1.In vitro differentiation of human skin-derived mesenchymal stem cells into lymphocytes: Possibility evaluation
Liping GUAN ; Jie YU ; Bing HUANG ; Ting LUO ; Jianfa HUANG ; Qian LIU ; Liping LIN ; Min ZHANG ; Kaijing LI ; Xigu CHEN
Chinese Journal of Tissue Engineering Research 2010;14(19):3601-3605
BACKGROUND: Previous research has demonstrated that dermal tissue has mesenchymal stem cells, which have a possibility of autologous transplantation. If the mesenchymal stem cells derived from the skin differentiate into lymphocytes under a certain condition, the immune system disease can be solved generally.OBJECTIVE: To investigate the possibility of differentiation of human skin-derived mesenchymal stem cells into lymphocytes. METHODS: Surface marker expression was detected in the 14th passage human skin-derived mesenchymal stem cells using flow cytometry. Transdifferentiation medium of human skin-derived mesenchymal stem cells consisted of human lymphocyte supernatant and fresh human skin-derived mesenchymal stem cells based on the ratio of 7:3. Inverted microscope was employed to observe morphological changes, and flow cytometry was used to detect surface marker expression in the lymphocytes at 1-8 days after induction. Self-marker expression of human skin-derived mesenchymal stem cells was then detected at 3,6, and 9 days after induction.RESULTS AND CONCLUSION: Human skin-derived mesenchymal stem cells stably expressed self-specific marker CD73, Vimentin and so on, but did not express specific markers of hematopoietic system, I.e., CD34, CD45 and so on, lowly expressed HLA-I, but did not express HLA-DR at all. At 3 days after induction, the cell volume significantly increased, cell proliferation rate was significantly lower than before induction, and a lot of cystic-like particles with strong refraction were observed in or between cells. The CD45 lymphocyte expression was not significantly changed, but CD3, CD19, CD16, CD4, and CD8 expression rates of human skin-derived mesenchymal stem cells were linearly increased at 1-4 days after induction and stabilized at 5-8 days after induction. In addition, CD37, CD34, Vimentin, and HLA-DR expressions were not changed at 3, 6, and 9 days after induction, but HLA-I expression rate was gradually increased with the prolongation time of induction. This suggested that human skin-derived mesenchymal stem cells can differentiate into lymphocyte and potentially participate in repairing immune system injury.
2.In vitro EdU labeling of peripheral blood mononuclear cells in rabbits
Minglei ZHAO ; Dongqin ZHEN ; Jianfa HUANG ; Weihua LI ; Wencong WANG ; Zhiquan LI ; Hening ZHANG ; Bikun XIAN ; Yuting PENG ; Minyi ZHOU ; Bing HUANG
Chinese Journal of Tissue Engineering Research 2017;21(9):1432-1438
BACKGROUND: The proliferation of peripheral blood stem cells among peripheral blood mononuclear cells (PBMCs) invitro remains unclear. There is no optimal marker for tracing PBMCs transplanted in vivo.OBJECTIVE: To observe the degree of PBMC proliferation in stem cell medium by EdU labeling and to explore thefeasibility of EdU-labeled peripheral blood stem cells.METHODS: New Zealand rabbit PBMCs were isolated and cultured for 1 to 5 days in stem cell medium supplementedwith EdU. The cells were observed and counted at 0, 1, 2, 3, 4 and 5 days in culture. The cells were harvested at eachtime point and stained with EdU fluorescent reagents. Then, confocal microscopy and flow cytometry were used to detectEdU-labeled cells.RESULTS AND CONCLUSION: (1) Freshly isolated rabbit PBMCs were rounded and showed clear outline. After 1 dayculture, most of the cells were suspended in the medium, spherical or round. There were also a few cell clusters andadherent cells scattered in a triangle or polygon shape; after 2 days culture, more cell debris were observed, and mostcells were round; when cultured for 3-5 days, increased cell debris, smaller cell mass and decreased cell densitysignificantly were observed. (2) With the prolongation of culture time, the cell count decreased gradually. (3) Whencultured for 1 day, EdU labeled cells in red were scattered. The number of cells marked with EdU red label increasedsignificantly at day 2 and remained unchanged after 3 days of culture. At 5 days of culture, the number of red cellsmarkedly decreased; the highest positive rate of EdU-labeled cells was (2.38±0.10)% at 2 days after culture. To conclude,these results showed that the proportion of proliferating cells in rabbit PBMCs was very low. EdU is capable of labelingproliferative cells among PBMCs.
3.Synchrotron radiation phase contrast CT imaging of in vitro biological soft tissue
Jianfa ZHANG ; Xiujun CHEN ; Wenjing LI ; Zhongjie HUANG ; Chaosen ZHONG ; Guangzhao ZHOU ; Shunfa LIN
Chinese Journal of Medical Imaging Technology 2018;34(2):171-175
Objective To explore the feasibility of in vitro biological soft tissue imaging by using synchrotron radiation phase contrast CT.Methods Three samples of resected human cardia,two samples of resected human esophageal carcinoma and esophagus,as well as two samples of middle cerebral artery tissue extracted from corpses were fixed and airdried at room temperature for synchrotron radiation phase contrast CT imaging.The images of soft tissue structures were observed and compared with pathological findings.Results The images of synchrotron radiation phase contrast CT showed three-layer structure of cardia and esophagus,mucous,submucosa and muscular layer.The surface of mucous layer was smooth.The images of esophageal carcinoma showed cancerous tissue infiltrating esophageal wall.The wall and lumen of cerebral arteries could be also clearly displayed.Conclusion Synchrotron radiation phase contrast CT imaging can clearly display fine structures of in vitro biological soft tissue.
4.The value of blood coagulation function in early stage of acute renal fail-ure
Min HE ; Jianwu QIU ; Jieping HUANG ; Xingjiang DENG ; Jun WU ; Jianfa HE
China Modern Doctor 2014;(18):163-165
Objective To study the changes of blood coagulation function in early stage of acute renal failure patients, and illustrate its significance. Methods From October 2011 through January 2013, 80 patients with acute renal failure in the Nephrology Department and Pediatrics Department of Yuebei People’s Hospital were enrolled in this study,50 health people in the same period as control group,then compare the differences in creatinine, blood urea nitrogen, blood platelet(PLT), activated partial thromboplastin time(APTT), prothrombin time(PT), thrombin time(TT), plas-ma fibrinogen (FIB) between the two groups. Results The FIB levels and PLT count in observation group were higher than those of controls group (P<0.05), but APTT,PT,TT showed no statistically significant difference (P>0.05). Con-clusion Hypercoagulabale state could occur in patients with acute renal failure in early stage, abnormal clotting may be participates in occurrence of acute renal failure.
5. Preliminary efficacy analysis of Cheng's Giraffe reconstruction after proximal gastrectomy in adenocarcinoma of esophagogastric junction
Xiangdong CHENG ; Zhiyuan XU ; Yi′an DU ; Can HU ; Jianfa YU ; Litao YANG ; Ling HUANG ; Pengfei YU ; Gaiguo DAI ; Yanqiang ZHANG
Chinese Journal of Gastrointestinal Surgery 2020;23(2):158-162
Objective:
To investigate the safety and feasibility of proximal partial gastrectomy with Cheng's Giraffe esophagogastric reconstruction for the treatment of early Siewert II adenocarcinoma of esophagogastric junction (AEG).
Methods:
Indication of Cheng's Giraffe esophagogastric reconstruction: (1) Siewert II AEG or Siewert III AEG with diameter < 4 cm; (2) preoperative staging as cT1-2N0M0. A descriptive case series study was carried out. Clinical data of 34 patients with Siewert II AEG undergoing proximal partial gastrectomy and Cheng's Giraffe esophagogastric reconstruction at Department of Abdominal Surgery of Zhejiang Cancer Hospital and Department of Gastrointestinal Surgery, The First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine from February to July 2018 were retrospectively collected and analyzed, including 14 cases in IA stage, 11 cases in IIA stage and 8 cases in IIB stage. Brief procedure of Cheng's Giraffe esophagogastric reconstruction was as follows: Firstly, 12 cm long tubular stomach was formed by longitudinal incision 4 cm away from the great curvature of the stomach. Secondly, the gastric fundus and His angle were formed. Finally, the distance from His angle to esophagal-tubular gastric anastomosis should be more than 5 cm. The reflux disease questionare (RDQ) scores, radionuclide gastric emptying scintigraphy, and 24-hour multichannel intraluminal (MII)-pH monitoring technology were used to evaluate postoperative gastric emptying and gastroesophageal reflux.
Result:
All 34 patients successfully completed proximal partial gastrectomy with Cheng's Giraffe esophagogastric reconstruction, including 13 cases by open surgery and 21 cases by laparoscopic surgery. The operation time was (144.6±39.8) minutes, the blood loss during operation was (35.4±17.2) ml. No laparoscopic case was converted to open surgery and no postoperative complication was observed. The postoperative hospital stay was (8.4±2.5) days. The postoperative RDQ score was 4.4±3.1 one month after operation, and 3.3±2.5 six months after operation. Gastric-half emptying time was (67.0±21.5) minutes, and the residual ratio was (52.2±7.7)% in 1 hour, (36.4±3.1)% in 2 hours and (28.8±3.6)% in 3 hours at postoperative 1-month. The 24-hour MII-pH monitoring at postoperative 2-month revealed the frequency of acid reflux was (12.6±7.9) times, frequency of non-acid reflux was (19.6±9.7) times, DeMeester score was 5.8±2.9.
Conclusion
Cheng's Giraffe esophagogastric reconstruction is safe and feasible in the treatment of Siewert type II AEG, and has good dynamic and anti-reflux effects.