1.僅用經驗性的理論對燒傷濕性醫療技術作出解釋是不夠的。
      20世紀90年代初，徐榮祥完成的燒傷濕性醫療技術在燒傷臨床治療醫學領域掀起了一場風暴。他的醫療技術徹底地改變了燒傷治療醫學的現狀，通過使用他獨創的藥物MEBO和濕性療法，即使是重度燒傷，患者的創面也能實現無疤痕癒合。
      革命性的治療效果對傳統的以燒傷外科為基礎的燒傷技術產生了強烈的震撼。徐榮祥和他的醫療技術成為傳統燒傷醫學爭論的焦點。一些學者否定濕性醫療技術，指責徐榮祥是騙子，其所謂的醫療技術是騙術。而另一些學者則不僅把濕性醫療技術看成是燒傷醫學的最高成就，自己也成為濕性醫療技術積極的鼓吹者和實踐者。
      同樣都是傳統燒傷外科的學者，針對同樣一個技術，態度和立場截然相反，針鋒相對，這是為什麼？
      反對者所以反對，支持者所以支持，實際上，都與一個核心問題相聯繫，那就是基理。
      反對者在看待和評價濕性技術時，是從經驗出發，以傳統的理論作為依據的。按照傳統的經驗，受損傷的皮膚組織除表皮可以自行生理性癒合而不留疤痕外，深層皮膚是不可能自行生理性癒合不留疤痕的。傳統的理論為這一經驗提供了解釋，表皮皮膚創傷可以癒合是因為表皮有基底幹細胞存在，當表皮受損，基底幹細胞會自然啟動，分化形成組織，並最終完成損傷皮膚的修復。即便皮膚未受損傷，正常的表皮組織代謝也需要產生新的組織以替代舊的組織，這一過程也是由基底幹細胞自然完成的。基底幹細胞主要位於表皮層，真皮層也有少量存在，因此，一旦燒傷深及表皮下的真皮、脂肪層或肌肉層，表皮的修復就不再可能，原因很簡單，那裡沒有幹細胞。根據這一既有的理論，一些學者自信地得出結論：隨便徐榮祥瞎吹，治好了這個，治好了那個，肯定是假的，不符合生命規律的事情，誰也辦不到！
      儘管經歷了近十年的發展，利用濕性醫療技術救治成功的病人累計已近上百萬，這一實際存在的現實卻仍不足以改變持反對意見者的態度。不能怨人食古不化，也不能怨人冥頑不化，問題在於基礎理論方面。在分子醫學的時代，人們已經能夠在分子水平上對一些影響生命的現象的規律作出更深刻更精確的描述，而濕性醫療技術仍僅僅通過在治療學方面總結出的一些經驗性的理論來解釋燒傷濕性醫療技術，這顯然是不夠的。
      不過，在支持徐榮祥濕性醫療技術的學者那裡，他們卻為濕性醫療技術受到的指責抱不平。從現實出發，在這些學者眼裡，事實是第一位的，理論是第二位的。徐榮祥濕性醫療技術的貢獻絕不單純是將燒傷醫學提高到一個前所未有的水平，更重要的是他通過自己的醫學實踐揭示了一種現象，即深度損傷的皮膚也是可以實現生理性癒合的。這不僅是治療醫學的革命，更是人類認識的一場革命。如果沒有濕性醫療技術，人類對於有關皮膚組織生長的認識不要說在本質上有多深刻，就是在現像上也是不全面的。
      對於濕性醫療技術，理論上講不清，並不是濕性醫療技術的錯，而恰恰是傳統的理論隨著新的現象的出現，已不能滿足現實的需要，現實要求它必須積極地正視現實，迎接挑戰，面臨更新。一些敏感的學者強烈地感到啟動濕性醫療技術基理方面的研究已迫在眉睫，勢在必行，同時他們也感悟到濕性醫療技術基理的研究對未來的影響，不會單純地局限在燒傷醫學的領域，它的效應可能會是一場核裂變，波及整個醫學，深及未來的生命科學。這或許也就是巴巴拉和高登兩位美國學者用諾貝爾獎作尺度來衡量其研究價值的真實原因！
      對於濕性醫療技術基理的研究，他的發明者徐榮祥更是孜孜以求。在濕性醫療技術誕生後，徐榮祥在進一步完善技術的同時，便開始了燒傷濕性醫療技術所實現的損傷皮膚無疤痕癒合基理方面的研究，其探索的努力也從未停止過。早在1989年，在徐榮祥創辦的《中國燒傷創瘍》雜誌的創刊號中，他和他的研究者就刊登文章研究在光學顯微鏡下觀察到的皮膚細胞表現特殊的生長情況。大規模地開展研究則是從1991年的最後一個季度開始的。
      2.幹細胞大兵團遠征計劃中途夭折
      1991年10月25日，美國新澤西海肯塞醫療中心著名燒傷醫學專家巴巴拉打來國際長途，興奮地向徐榮祥通報了他對燒傷濕性治療技術的核心藥物BC4D進行重複實驗研究的情況。以下是當時電話記錄的摘要：
      1990年12月你來美訪問，臨走時，交給我的關於MEBO研究，經過一年的實驗，終於有了結果，你留下的MEBO共進行瞭如下實驗，結果很令人滿意。
      第一個實驗是利用MEBO進行了體外細胞的培養研究。簡單說，實驗方法如下：用兩組試管，均裝有細胞培養液並放入上皮細胞，然後在其中一組中的試管液內放入MEBO，而另一組不放入MEBO，進行對比細胞培養試驗。其實驗結果是非常令人吃驚的。發現在放入MEBO的試管中，其上皮細胞變為基底層細胞，且生長速度非常快，其高速生長的基底層細胞在質量上均是成熟的。這一切我們均利用較精密的儀器進行了測量。這樣的結果是在國際上的重大突破。在世界上，任何藥物均不能出現此結果！
      第二個實驗結果：將成塊真皮放入細胞培養液中培養，發現殘留在真皮內的殘存毛囊腺上皮及毛囊腺在MEBO的作用下，再生速度非常快，再生出的毛囊腺、毛囊器的形態與正常原有的相同。這一切變化均是通過顯微鏡而直接觀察到的。對這樣的變化，以前人們曾根據毛囊的再生規律預想過，但因毛囊上皮生長速度極為緩慢，從沒有人能描述，更不能直接觀察到毛囊再生的情況，而使用MEBO後，毛囊腺上皮再生速度明顯加快，人們可直接觀察到。這是一個重要的國際性突破。
      第三，關於分子生物學的研究結果：我們對使用MEBO後的組織細胞的再生基因工程進行了研究，通過實驗儀器對RNA、DNA合成了測量。實驗結果證明：使用MEBO後，其中的DNA、RNA的數量大量增加，說明細胞的分裂速度較快。這一結果和以上所報告的結果都是極為重要的，在世界上也從來沒有人做過此項試驗，也從來沒有獲得這樣的結果。
      這些結果對美國FDA通過MEBO進入美國市場也有著更重要的作用。這項實驗是在美國FDA認可的實驗室內進行的，是完全可信的，更何況此項實驗是FAD撥款的。
      儘管巴巴拉描述的這些實驗中的一些實驗，徐榮祥已經做過，但是從大洋彼岸傳來的信息還是使徐榮祥產生了一種競爭世界科學最前沿的緊迫感。放下電話，徐榮祥匆忙整理完材料，便直奔衛生部科技司，向有關領導作了匯報，並提議由國家主持正式全面啟動濕性醫療技術有關基理的研究。徐榮祥的建議立即得到衛生部科技司和部主要領導的高度重視。
      1991年10月30日，在衛生部科技司召集和主持下，一個由中國醫學科學院、中國預防醫學科學院和北京醫科大學等權威醫學研究機構的基礎研究專家參加的大型學術報告會在首都賓館舉行。會上，徐榮祥向基礎醫學專家詳細地介紹了燒傷濕性醫療技術對燒傷皮膚組織實現生理性修復所起到的特殊功效，並提出了基礎研究所要研究的課題。這些課題包括：一、汗腺上皮再生表皮，二、真皮及附件完全再生，三、DNA高速變化規律。
      徐榮祥的報告激活了基礎醫學專家的神經，僅僅一個月，近五十名著名的基礎醫學專家提交了落實研究的詳細課題設計，直接參加這項研究的科研人員多達150人。當厚厚的課題報告書堆滿了案頭的時候，徐榮祥彷彿進入了遠古的英雄時代，他像史詩中的英雄一樣真實而深切地感受到了一場世紀之戰的恢宏。
      多麼雄渾激越的時代，多麼催人奮進的計劃，多麼激動人心的研究！
      正當圍繞揭示皮膚細胞生長規律而展開的大兵團作戰的英雄史詩的帷幕即將拉開的時候，徐榮祥突然受到了來自傳統燒傷醫學最猛烈的衝擊。
      儘管在衛生部的關懷下，在國務院的支持下，徐榮祥躲過了劫難，沒有被狂風暴雨所襲倒，但是他剛剛組織起來的遠征軍卻被支解得七零八落。在以後的日子裡，抨擊與反抨擊、調查與被調查、陷害與反陷害的角逐構成了徐榮祥人生中一段複雜而險越的經歷。為了挺直燒傷濕性醫療技術稚嫩的身軀，他不得不捲入一場求生的鬥爭，皮膚組織細胞學的研究計劃被迫暫時埋藏在了心靈的最深處。
      1994年底，徐榮祥的命運引起剛剛走馬上任的衛生部新任副部長張文康的關注，他找到徐榮祥，關切地詢問了濕性醫療技術有關基理的研究情況。當得知徐榮祥的研究因學術紛爭而裹足不前時，張文康當即果斷地指示，研究工作不能中止，衛生部將努力創造良好的環境，支持徐榮祥的研究。
      躲過了寒冬的徐榮祥，倍感春天的溫暖。1995年5月22日，通過中央電視台的《新聞聯播》，徐榮祥向社會公佈了啟動皮膚幹細胞的研究計劃。
      3.鎖定合成人類角蛋白19型細胞
      沒有大規模的兵團，沒有聲勢浩大的儀式，僅僅帶領了一支不到十人的隊伍，徐榮祥神秘地從公眾媒體上銷聲匿跡。1996年4月1日，他帶領著自己的研究小組離開北京，驅車直奔有大量正在接受濕性醫療技術治療的重度燒傷病人的湖北襄樊，開始了他有關皮膚再生基理的細胞學臨床與基礎密切結合的研究。
      徐榮祥這一次開展研究同5年前組織大兵團時在思路發生了很大的變化。5年來為求生存而展開的鬥爭固然耽誤了許多寶貴的時間，但是時間也使徐榮祥的思想得到了一次沉澱的機會。從對細胞的狂熱到冷靜，時間濾去了一些浮躁的表皮的虛像，而使得隱秘的本質的聯繫漸漸地凸顯出思維的平台。
      5年間，徐榮祥並未停止對皮膚組織再生基理的思考，最初這些思考呈現為一種靈感式的迸發狀態，思想的火花五彩繽紛，四處飛濺，豐富而零亂，彷彿急需開展的研究有很多很多，簡直是五花八門，樣樣齊全，專家申報了50多個課題仍讓他感到不夠過癮。而隨著時間的推移，漸漸地，他的思想開始完全集中到了乾細胞這唯一的目標上。
      他的思想被一個在細胞學上稱為角蛋白19型的干細胞牢牢地控制著。最初，並沒有通過邏輯性演繹縝密清晰的步驟，僅僅是一個靈感，一種直覺使他關注到這種細胞。他及時地捕捉住它，生怕它會從裂變的思維中消失。後來，通過理性的分析，在沒有得到實驗證實之前，他就已作出了超前的判斷：完成深度損傷皮膚生理性修復的基礎物質很可能就是人類角蛋白19型這種干細胞。這一判斷一經作出，他的大腦就被完全鎖定了，幾乎到了想掙脫都沒辦法掙脫的地步。
      早在1989年，徐榮祥就通過光學顯微鏡直接觀察到燒傷的皮膚組織在接受MEBO這一藥物治療後細胞組織出現的特異性變化。這和巴巴拉在體外觀察的結果是相同的。在施用了MEBO之後，燒傷創面上增生了許多細胞，這些細胞聚集在一起形成了一個群狀的細胞團。這樣的變化在沒有用藥的創面上是根本見不到的。當徐榮祥把這樣的組織切片送到解放軍總醫院細胞室進行觀察的時候，權威的細胞學專家不禁驚嘆：“這是什麼組織，燒傷的皮膚創面怎麼會有胚胎組織類型的細胞！” 
      根據已知的基礎研究提供的有關皮膚組織細胞的知識，徐榮祥知道已探明的皮膚分化程度不同的表皮細胞含有5種類型的角蛋白：合成人角蛋白1型、10型、9型、16型和19型。這些不同類型的細胞其功能表達各不相同。合成1型和10型角蛋白的細胞是生成較硬皮質的細胞，它是成熟的表皮細胞，其分化增生的功能已接近完結；合成角蛋白9型和16型的細胞是表皮幹細胞移行過程中的中間型細胞，是可以分化形成合成1型和10型角蛋白的細胞，屬於有分化能力的未成熟細胞；而含角蛋白19型的細胞則是提供表皮再生的，分化能力最強的表皮幹細胞。
      完成人類皮膚的生長和修復，這些細胞在不同階段都起到了相應的作用，但是它們中除了19型是最原始的類型外，其餘的1型、10型、9型和16型都是中間階段的類型，它們本身就存在於表皮層，在表皮以下的組織中是不存在的。因此要實現深度燒傷皮膚修復，這樣類型的細胞是力所難及的。唯一有可能的只有19型。
      不過，要讓思維確信無疑地接受19型，並不是順理成章的事，中間仍然有思想障礙需要排除。因為19型是胚胎細胞，也就是說根據已有的知識，它只有在人剛剛由受精卵在子宮中發育成胚胎時才可以見到，隨著胚胎進一步的發育，19型就會分化變成其他類型的細胞。如果要確定19型，那麼，除非首先要在19型細胞可能出現的位置上實現突破，即在被認為是不可能出現的地方，如表皮層、真皮層、脂肪層，甚至肌肉層中尋找到19型的踪影；其次，就是跟踪19型的變化規律，如果在深度燒傷皮膚癒合的過程中，能夠確定地追踪到19型細胞分裂、增生，並有轉化為其他更為成熟的諸如1型、 10型、9型和16型等類型的跡象的話，確定19型才可以說是板上釘釘的定論。
      4.拿下乾細胞有如探囊取物
      理論先行使實踐免走了很多的彎路，原來認為需要瞄準多個目標，進行大規模協同作戰的戰略，現在變得沒有必要。現在的目標非常明確：就是要直截了當拿下合成人角蛋白19型細胞。原本要動用千軍萬馬的巨工程被簡化成僅僅只要一個科研小組就可以完成的工作。而這個科研小組所作的工作也被壓縮成兩個方面。一方面是嚴格監督臨床治療的規範性，以便按照根據對照取樣需要設定的時間表，準確地從患者創面獲取到可供基礎研究觀察的病理切片。另一方面是選擇最優秀的實驗室，用最先進的細胞追踪技術，追踪幹細胞的變化規律。
      正巧這時湖北襄樊有一批重度燒傷病人，他們為開展幹細胞的基礎實驗研究提供了得天獨厚的機會。徐榮祥親自參與搶救和治療每一個病人，並指導研究人員按設計規範做病理切片。同時，他還不得不時常趕回北京，與中國醫科大學的實驗人員共同探討實驗設計和方法。有一次，當他回到北京，剛下飛機打開手機，襄樊臨床基地的電話便追了過來。一位燒傷危重病人突然病危，臨床醫生處理不了。徐榮祥毫不遲疑，沒出機場，立即又登上了返程的航班。就在當地醫生作出決定，準備對這位病人放棄治療時，他衝進了病房。經過他簡單的處理，病人的危險症狀立即得以緩解。 
      北京方面的研究情況也曾出現過周折。最初，實驗人員對切片進行免疫熒光觀察時，結果很不理想，在切片上，他們沒有發現有合成角蛋白19型的細胞。當這樣的結果呈送到辦公桌上的時候，徐榮祥感到很意外，他根本不相信會是這種結果，因為他對19型很有信心。
      無論從純理論的推斷，還是從實驗觀察所使用的技術方面都很難找出缺陷，徐榮祥開始對實驗的結果進行分析，這時他發現實驗結果反映出的情況有些離譜，切片反映出的細胞狀態是發生傳代變異了的，而不是原始的。這說明取切片的方法可能存在問題。
      回過頭來，重新檢討取切片的方法，徐榮祥發現產生他推斷的那種可能性確實存在，於是他和驗證課題組的成員中國醫學科學院協和醫科大學的許增祿教授商議決定對切片進行即時冷凍並置於液氮中保存。保存切片的方法一改變，實驗觀察便迅即走上了一條通衢大道。
      接下來的工作就是對切片進行生物素抗生物素蛋白DCS體系間接免疫染色。生物素抗生物素蛋白DCS體系間接免疫染色是目前世界上最先進的細胞染色技術，它使目標能夠通過熒光顯示得到特異性的標識，因而是追踪細胞變化的理想手段。
      首先，為了測定接受治療的燒傷皮膚組織中確實存在角蛋白19型細胞，研究者選用小鼠抗人角蛋白19型的單克隆抗體進行測試。實驗同時對正常皮膚與接受濕性醫療技術治療的燒傷皮膚進行對照測試，以觀察二者有無特異反應。實驗結果是理想的，研究者在正常皮膚組織中沒有發現有呈陽性反應的角蛋白19型細胞，而在接受MEBO藥物治療的燒傷皮膚組織中，24小時後，發現有顯示熒光呈陽性反應的角蛋白19型細胞。
      其次，繼續觀察接受治療的燒傷組織細胞的變化情況。研究者發現治療後4天，在汗腺、毛細血管和毛囊周圍潛在的角蛋白19型細胞開始增多。治療後7—14天，角蛋白19型細胞的數量接近並達到峰值。至21天和28天這個時間段，角蛋白19型細胞的數量開始回落。這個觀察結果與燒傷皮膚接受治療的臨床及組織學研究結果正相吻合。
      這項研究實現了研究者最初設定的目標，即確定角蛋白19型乾細胞，追踪其分化增生的規律。燒傷皮膚組織再生修復的基理終於顯出了端倪。
      第一，通過這項研究證實了在殘存的燒傷組織中可產生人角蛋白19型這種最原始的胚胎類型的細胞；人角蛋白19型細胞數量上的變化說明人角蛋白19型細胞具有不斷分化和持續增殖的功能。
      第二，從免疫熒光染色特異性表達的結果可以發現它在移動和分裂後，能產生合成其他類型角蛋白1型和10型的細胞，這些細胞繼續分裂產生合成成熟表皮細胞所含的典型的角蛋白1型和10型的細胞，這充分說明人角蛋白19型細胞就是承擔燒傷皮膚組織再生修復的細胞。
      第三，研究者還初步確定了人角蛋白19型細胞啟動調控的規律。研究表明，細胞是從靜止期釋放出來的，首先被激活的周期蛋白是cyclin D，在受到生長因子刺激後，它才表達，真核生物在細胞的G1期決定進入增生狀態還是退出週期，此時主要的調控者是周期蛋白cnclin D1/CDK4複合體，由此可以證實濕性醫療技術對乾細胞分裂複製和再生具有啟動和調控的特殊作用。
      第四，研究中還尋找出了成纖維細胞、血管組織細胞及皮膚神經細胞在皮膚再生過程中各自及相互間的持續增殖及形成皮膚的規律，其詳細成果研究者將陸續公佈。
      1998年4月，徐榮祥領導的科研小組旨在確定損傷皮膚生理性癒合基理的干細胞研究已初步完成，人類試圖利用乾細胞治療臨床疾病的宏圖首先在古老的東方美夢成真！

1. To explain MEBT (Moist Exposed Burn Therapy) techniques by empirical theory itself was not enough and satisfactory.

In the early 90s of 20th century, Rongxiang Xu completed his MEBT techniques, which shocked the clinical burn society.  His therapeutic techniques had entirely changed the current state of burn treatment.  By applying MEBO, his uniquely invented medication, and moist techniques (MEBT), wound healing without scar formation could be achieved even in severe burn cases.

These revolutionary therapeutic effects strongly shocked the traditional burn treatment techniques based on burn surgery.  Rongxiang Xu and his medical techniques became the center of debate in traditional burn medicine in China.  Some researchers denied the MEBT techniques, blamed Rongxiang Xu as a liar, and his techniques as a scam.  On the other hand, other researchers believed that the MEBT techniques were the most advanced achievements in burn medicine.  They became advocates and applied MEBT techniques themselves.

Why, among the same group of traditional burn medicine researchers, was there a totally contradicted opinion towards the same technique?

The key question based on which the opponents opposed, or the advocates supported the techniques, was about the mechanisms behind the techniques.

The critics against the moist techniques were based on experiences and traditional theories. In accordance with experiences, with the exception of epidermis that could automatically undergo physiological healing without scar formation after injury, injured deeper skin tissues were not able to heal without scar by themselves.  The rationale provided by traditional theory was that the basal stem cells residing in epidermis contributed to epidermal healing after epidermal injury.  When the epidermis was injured, basal stem cells were automatically initiated to differentiate into tissues to repair.  Even without injury, the old tissues needed to be replaced by new tissues during normal epidermal metabolism, which was carried out by basal stem cells also.  Basal stem cells were mostly located in epidermis, with a few found in dermis.  As a result, epidermal repair became impossible once the burn was deep into dermis, fat tissues, or underlying muscles where there were no stem cells.  Based on this established theory, some researchers confidently concluded: Whatever Rongxiang Xu claimed by his wish that he can cure, it was not real.  Nobody can make achievement against the principles of life. 

The fact that during the nearly ten years of development, MEBT had successfully treated millions of patients was still not enough to change the opponents’ attitude.  Their stubbornness should not be blamed, since the key lay in the fundamental theory.  In the time of molecular medicine, the principles of life could be thoroughly and accurately elucidated on molecular level, whereas the approach of moist therapy tried to explain the mechanisms by empirical theory, which was clearly equivocal.

Nevertheless, the supporting researchers, who believed that the fact is the first priority and the theory is only the second, felt the techniques were unfairly treated.  The contribution of Rongxiang Xu’s MEBT was not merely raised burn medicine to a level never seen before; more important, it revealed through clinical practice that deep injured skin is able to undergo physiological healing.  This was a revolution not only in therapeutic medicine, but also in human recognition.  Without moist therapy techniques, our understanding on skin tissue generation would not be intrinsically profound or superficially comprehensive.

MEBT could not be clearly explained in theory, which was not the fault of the techniques.  Quite the contrary, it was due to the insufficient understanding by traditional theories in new phenomena, which did not meet the needs in reality.  The reality required the tradition to confront the reality, the challenge, and the innovation.  Some researchers already sensed the imperative need to commence study on the mechanisms of MEBT techniques and recognized their significance for the future, which will not be limited in burn medicine.  The mechanisms would be a ‘nuclear fission’ involving the entire medical field and the future life science.  This was likely the reason that the two American researchers, Barbara and Gorden, used ‘Nobel Prize’ as the specification to assess the value of this study.

Rongxiang Xu, the inventor of MEBT techniques, had been diligently strived after the mechanisms behind MEBT for years.  After the birth of MEBT, Rongxiang Xu had further improved his techniques, and simultaneously initiated the study on the mechanisms behind skin wound healing without scar.  As early as in 1989, on the first issue of the journal The Chinese Journal of Burns Wounds & Surface Ulcers founded by himself, Rongxiang Xu and his research team published their research paper, in which the special cell growth observed under optical microscope was reported.  In the last quarter of 1991 the full scale research was ready to launch.

2. Expedition of stem cell army was aborted halfway through

On October 25th, 1991, Barbara, the famous burn specialist from Hackensack University Medical Center informed Rongxiang Xu through international phone conversation about the repeated experiments he performed on BC4D, the core medication of MEBT.  The following were the summary of this conversation:

“You visited America in December, 1990.  Before leaving, you gave me the study about MEBO.  After 1 year efforts, we finally got the results now.  By using MEBO you left, we performed following experiments, and the results are satisfying.

The first experiment was to use MEBO to study cell culture in vitro.  In short, the experimental methods were as follows: using two groups of epithelial cells in the test tubes all filled with media.  MEBO was added into only one group of test tubes (test group).  The cell culture results were then compared.  The results were remarkable.  In the test group, the epithelial cells transformed into mature basal cells with rapid growth, which was determined by precision apparatus.  These results, could not be obtained by using any other drug, were a significant world-class breakthrough!
 
The results of the second experiment: putting dermal explants into cell culture media with MEBO, which resulted in rapid regeneration of residual hair follicle gland epithelium and hair follicle gland under MEBO effects, the morphology of regenerated hair follicle gland and hair follicle appendage were unchanged.  All of these were observed directly through microscope.  Previously, people had only prefigured the similar change by the same pattern of hair follicle regeneration.  However, the growth of hair follicle epithelium was extremely slow so that nobody was able to describe, nor to observe the regeneration directly.  But, after MEBO application, considerable increase of the growth rates could be observed directly.   This was also a big international breakthrough.

The results of the third experiment regarding molecular biology: we tried to study the ‘regenerative’ gene engineering of regenerated tissue cells applied with MEBO, and measured the DNA and RNA with experimental tools. The experimental results showed: after the application of MEBO, the number of DNA and RNA in the tissue cells were significantly increased, which indicated rapid cellular division… The results, as important as those above, were never attempted and obtained before by anybody in the world.

These results had major impacts on the FDA approval of MEBO entering into American market.  Since the experiments were performed in FDA approved laboratory and even funded by FDA, they were totally reliable.”

Even though Rongxiang Xu had completed the experiments performed by Barbara, the information coming across the sea still brought the urgency to him.  After the phone conversation, Rongxiang Xu reorganized related files and went to the science department of China’s Ministry of Health.  He reported to the persons in charge and suggested to initiate full scale study under government supervision on the mechanisms of moist therapy techniques.  His suggestion drew immediate attention from both the department and the ministry.

On October 30th, 1991, directed by the science department of Ministry of Health, a large conference involving researchers in basic medical science from various research authorities, including Chinese Academy of Medical Sciences, Chinese Academy of Preventive Medicine, and Beijing Medical University, was held in Capital Hotel.  During the conference, Rongxiang Xu introduced the extraordinary effects of MEBT on physiological regeneration in burn skin tissues.  He also recommended the subjects need to be explored further, which included: 1) epidermal regeneration by sweat gland epithelium, 2) full regeneration of dermis and appendages, and 3) the study on the pattern of rapid DNA change.

The report by Rongxiang Xu genuinely encouraged these researchers in basic medical science.  Within one month, a detailed study design was prepared and submitted by nearly 50 prominent researchers in basic medical science of China, with up to 150 researchers involved.  Facing the thick folders of study description, Rongxiang Xu seemed going back into the ancient world, genuinely felt the greatness of the war of the century like an epic hero.

What a great era, what an encouraging plan, what an exciting study!

While the group study on the pattern of skin cell growth was underway, the battle began and the curtain was rising. Suddenly, Rongxiang Xu was heavily attacked by traditional burn medicine in China. 

Though Rongxiang Xu eluded from this adversity by the support from Ministry of Health and the State Council of China, his expeditionary army was dismembered.   During the following days, Rongxiang Xu experienced a series of complex and risky events consisted of critics and supports, investigating and being investigated, and frame and counter-frame.  He was forced to be involved in a struggle in order to back his MEBT techniques up.  The research plan of skin tissue cytology had to be buried deeply in his heart.

Dying before conquer, mourning ever since.

At the end of 1994, Rongxiang Xu’s experience received attention from the newly appointed Health Minister Wenkang Zhang, who inquired to Rongxiang Xu about the research status of the mechanisms in moist therapy techniques.  After acknowledging that Rongxiang Xu’s study was terminated due to the academic dispute, Wenkang Zhang resolutely instructed that the study could not be stopped; the Ministry of Health would create a healthy environment to support Rongxiang Xu’s research. 

After a cold winter, Rongxiang Xu was welcomed by the warmness of spring.  On May 22nd, 1995, he announced the initiation of skin stem cell research on Chinese CCTV Xinwenlianbo (CCTV network news).

3. To locked up ‘induced human keratin 19 positive cells’

Without a large-scale corps, without massive ceremony, Rongxiang Xu, along with his research team with less than 10 people, disappeared from press and public view.  On April 1st, 1996, he and his research team left Beijing and headed to Xiangfan, Hubei Province where there happened to be large numbers of severe burn patients receiving the treatment of MEBT techniques.  From there, his basic and clinical cytology combined study on skin regenerative mechanisms began.

This time his opinion about the study, comparing with five years ago, had changed dramatically.  Despite the time wasted in the struggle for the survival of his study in the last five years, the experience also gave Rongxiang Xu a chance to be composed.  From fervor to calmness, the impatience was removed by the time, and the hidden correlation of nature gradually emerged in his mind.

During the five years, Rongxiang Xu had never stopped thinking about the skin regenerative mechanisms.  Firstly, these thoughts surfaced as inspirational bursts, full of intellectual sparkles, splattering everywhere, rich and disorderly, just like the wide variety of more than 50 study subjects submitted by other scientists.  Gradually, his mind focused on only one subject---stem cell.

Rongxiang Xu’s mind was controlled by a cytological concept of keratin 19 stem cells.  Initially, he promptly noticed this cell not through logical deduction, but an inspiration.  Nonetheless, he kept himself focusing on this cell and was afraid that it might disappear from his fissile mind.  Later, through rational analysis, he predicted even before confirmed by experiments: the basic causative substances for the complete physiological repair of deep injured skin were most likely the keratin 19 positive stem cells.  Once the conclusion was drawn, his mind was locked in an unshakable state.

Back in 1989, Rongxiang Xu directly observed through optical microscope that after MEBO application, the cells in the burn tissues underwent specific changes.  This was consistent with the in vitro results obtained by Barbara.  Once MEBO was applied, large amount of cells proliferated in the burn wound and congregated to form a cell mass.  This change could not be seen in the wound without MEBO application.  When the tissue slide was sent to Chinese General Hospital of the People's Liberation Army, the authoritative cytologist exclaimed: ‘what is this tissue, how can embryonic tissue cells come to the burn skin wound?!’

Based on the knowledge of skin tissue cells provided by known basic research, Rongxiang Xu acknowledged that the detected skin epidermal cells in various differentiation states contain 5 types of keratin: human keratin 1, 10, 9, 16, and 19. Different cell types have different functional expression.  Cells containing positive keratin 1 and keratin 10 are mature epidermal cells with the ability to produce harder stromal cells, their proliferation and differentiation ability are almost exhausted; Keratin 9 and 16 positive cells are immature intermediate cells appeared in the migration process of epidermal stem cells with the ability to differentiate into keratin 1 and 10 positive cells; keratin 19 positive cells are epidermal regeneration cells with the most potential differentiation ability.

All of the above cells contribute to the growth and repair of human skin at different stages.  However, except keratin 19 positive cells, all other cells, including keratin 1, 10, 9, and 16 positive cells are intermediate cells naturally residing in epidermis only.  Therefore, none of these cells, but keratin 19 positive cells can complete skin regeneration in deep burn wound.

Still, there are mental barriers that make the unquestionable acceptance of keratin 19 more difficult than it should be.  Based on what we already know, keratin 19 positive cells are embryonic cells that can only be found in the process of embryo development from zygote in the maternal uterus.  By the development of embryo, keratin 19 positive cells will differentiate into other cell types.  To identify keratin 19 positive cells, firstly we must find the possible residence, i.e. try to find them in epidermis, dermis, fat layer, and even muscles, where are thought to be impossible for them to reside; in addition, we have to track the alteration and transformation of keratin 19 cells.  If we can identify the division, proliferation, and differentiation of keratin 19 cells into keratin 1, 10, 9, and 16 positive cells in the skin healing process in deep burn wound after MEBT, then we can be certain that the keratin 19 cells are found.

4. Conquering stem cell was easy as pie

Theory-first approach made the practice straightforward.  Previously it was believed that a full-scale, joint research with multiple subjects was essential to achieve the goal.  Now it seems not necessary.  The subject was simple and clear: to clarify the cells with keratin 19 positive expression.  This enabled a simple research unit to finish the study, which was originally thought a huge project and requiring substantial personnel resources.  The research unit was further divided into 2 teams.  One was to strictly supervise the clinical practice to make sure that in order to perform basic research observation, patients’ biopsy samples will be accurately obtained as scheduled.  The other team was to choose the best laboratory in which the transformation of stem cells would be followed by the most advanced cell tracking technology.

Sometimes destiny counts.  Coincidentally, there was a group of severe burn patients in Xiangfan, Hubei Province, which provided a valuable chance for basic stem cell research.  Rongxiang Xu himself participated in the rescue and treatment of each patient.  Furthermore, he guided researchers to make pathological slides in accordance with the planned specification.  At the same time, he had to travel back to Beijing once in a while to discuss experiment designs and methods with researchers from China Medical University.  In one occasion, right after arrived at Beijing airport and turn his mobile phone on, he received a phone call from the clinical center in Xiangfan, which informed him that there was a patient in critical situation and the hospital physician was unable to handle.  Rongxiang Xu immediately took another flight heading back without hesitancy.  Just when the hospital physician decided to give up on that patient, Rongxiang Xu arrived.  By his management, the life threatening symptoms in this patient was relieved soon.

There were also setbacks during the experiments in Beijing.  Initially, researchers found that the immunofluorescent results showing no keratin 19 positive cells expressed on the slides.  Rongxiang Xu was surprised by the results sent to his office.  Since he had unbreakable confidence on keratin 19, he did not believe the results.  There must be something wrong somewhere.  It was hard to find any error through either purely theoretical deduction, or the examination of the experimental procedures applied during the observation.  Rongxiang Xu started to analyze the experimental results and found that they were far from normal.  The cells on the slides showed mutation generated by passage and were not the original.  This indicated that there were errors in the sampling process.

Going back through the sampling methods, Rongxiang Xu found that what he proposed was possibly true. He discussed this with Zenglu Xu, professor of Chinese Academy of Medical Sciences affiliated Peking Union Medical University and a member of study subject validation team, and decided to immediately freeze the slides after sampling and reserve the slides in liquid nitrogen.  Once the storage condition was changed, the experimental observation became uncomplicated.

Next step was to perform indirect staining with biotin – avidin DCS system on the biopsy slides.  Indirect staining with biotin – avidin DCS system was the most advanced cell staining technique in the world.  It could show specific mark through fluorescencent display and therefore, was the ideal cell tracking method.

Firstly, in order to determine the existence of keratin 19 positive cells in the treated burn tissues, researchers selected a monoclonal antibody, mouse anti–human keratin 19 antibody as the tool.  During the experiment, the normal skin taken as control and was compared to the burn skin treated by MEBT, to see if there was any specific response.  Researchers found no keratin 19 positive cells in the normal skin, which met the anticipation.  On the other hand, keratin 19 fluorescent positive cells were found in MEBO treated burn tissues 24 hours after application.

In addition, observation on treated burn tissue cells continued.  Four days after treatment, researchers found increased number of keratin 19 cells in sweat gland, capillaries, and the tissues surrounding hair follicle.  Seven to fourteen days after treatment, the number of keratin 19 cells reached the peak.  Twenty-one to twenty-eight days after treatment, the cell numbers began to drop.  These results were consistent with clinical and histological results obtained from treated burn skin.

The experiment achieved the initially established objective to identify keratin 19 positive stem cells, and to track their proliferation and differentiation.  The mechanisms of burn skin tissue regeneration and repair emerged.

I. The experiment demonstrated that human keratin 19 positive cells, the most primitive embryonic cells, could be produced in the residual burn tissues; the change of the numbers of keratin 19 positive cells indicated that these cells had the ability to continuously differentiate and proliferate;

II. The results from immunofluorescent stain showed that keratin 19 positive cells could, after migration and division, generated other types of keratin positive (keratin 1 and 10) cells contained in mature epidermis, which sufficiently proved that burn skin tissue regeneration and repair were carried out by these human keratin 19 positive cells;

III. Researchers also primarily confirmed the initiation and regulation model of human keratin 19 positive cells.  Research showed that cells were released from stationary phase.  The firstly activated protein was cyclin D, which was expressed only by the stimulation of growth factors.  The cells in eukaryotic organisms were regulated by cycle protein cyclin D1/CDK4 complex at G1 phase to determine whether to enter proliferative state or withdraw from the cycle.  Thus, the initiative and regulatory effects of MEBT techniques on stem cell division and self renewal, and regeneration was proved.

IV. Researchers also discovered that fibroblasts, vascular tissue cells, and skin neural cells were able to continuously proliferate and generate skin by either themselves, or by the interaction between them.  The detailed results would be gradually published in the future.

In April, 1998, the research team led by Rongxiang Xu primarily completed the study on identifying the mechanisms in physiological healing of injured skin after treated with MEBT/MEBO.  The beautiful dream of treating clinical diseases with stem cells was fulfilled in the ancient east.