BACKGROUND:There are many kinds of cell media with different components,which have a great influence on cell growth.Several studies in and outside China have used serum-free media containing fetal bovine serum for in vitro amplification culture,but the use of media containing human peripheral blood serum to directionally induce human umbilical cord mesenchymal stem cells to neural stem cells and human peripheral blood serum to promote neural stem cells to differentiate into other nerve cells,so far there are few relevant studies. OBJECTIVE:To observe the feasibility of inducing human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells. METHODS:(1)Human umbilical cord mesenchymal stem cells were cultured in DMEM/F-12 culture medium containing 10%human peripheral blood serum by volume fraction.Flow cytometry analysis was performed at the third passage to identify surface markers and alizarin red staining was used to detect osteogenic differentiation function.(2)The third-generation human umbilical cord mesenchymal stem cells were induced into neural stem cells using DMEM/F-12 medium containing 0.5%N2,1.5%B27,20 ng/mL basic fibroblast growth factor,and 20 ng/mL epidermal growth factor,and their surface markers were identified.(3)Well-growing human umbilical cord mesenchymal stem cell-derived neural stem cells were taken to prepare a single cell suspension.They were evenly inoculated into 96-well plates and incubated with DMEM/F-12 culture medium containing 10%human peripheral blood serum for 8 days.Then,hematoxylin-eosin staining,microtubule-associated protein 2,and glial fibrillary acidic protein immunofluorescence staining were performed to detect the differentiation of neural stem cells into other neural cells. RESULTS AND CONCLUSION:(1)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum grew in a spiral-like pattern and were distributed in multiple layers,with directional arrangement.The surface of human umbilical cord mesenchymal stem cells highly expressed CD44,CD105,CD29,and CD73.Cells stained with alizarin red showed a color reaction.(2)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum could be induced to differentiate into neural stem cells,and the surface of neural stem cells was highly expressed with CD44,CD105,CD29,CD73,Nestin,NF-L,and GALC.(3)On day 8 of induced differentiation of neural stem cells,after staining with hematoxylin and eosin,it was found that the protruding protrusions were longer,with more branches and adjacent cells connected,presenting typical neural cell morphology.Immunofluorescence staining for microtubule-associated protein 2 and glial fibrillary acidic protein was positive.It is concluded that human umbilical cord mesenchymal stem cells cultured by human peripheral blood serum can be directly induced to differentiate into neural stem cells.Under the action of human peripheral blood serum,neural stem cells can differentiate into other neural cells as the culture time prolongs.

Objective:To evaluate the safety of double and a half layered esophagojejunal anastomosis in radical gastrectomy.Methods:This prospective, multi-center, single-arm study was initiated by the Affiliated Cancer Hospital of Zhengzhou University in June 2021 (CRAFT Study, NCT05282563). Participating institutions included Nanyang Central Hospital, Zhumadian Central Hospital, Luoyang Central Hospital, First Affiliated Hospital of Henan Polytechnic University, First Affiliated Hospital of Henan University, Luohe Central Hospital, the People's Hospital of Hebi, First People's Hospital of Shangqiu, Anyang Tumor Hospital, First People's Hospital of Pingdingshan, and Zhengzhou Central Hospital Affiliated to Zhengzhou University. Inclusion criteria were as follows: (1) gastric adenocarcinoma confirmed by preoperative gastroscopy;(2) preoperative imaging assessment indicated that R0 resection was feasible; (3) preoperative assessment showed no contraindications to surgery;(4) esophagojejunostomy planned during the procedure; (5) patients volunteered to participate in this study and gave their written informed consent; (6) ECOG score 0–1; and (7) ASA score I–III. Exclusion criteria were as follows: (1) history of upper abdominal surgery (except laparoscopic cholecystectomy);(2) history of gastric surgery (except endoscopic submucosal dissection and endoscopic mucosal resection); (3) pregnancy or lactation;(4) emergency surgery for gastric cancer-related complications (perforation, hemorrhage, obstruction); (5) other malignant tumors within 5 years or coexisting malignant tumors;(6) arterial embolism within 6 months, such as angina pectoris, myocardial infarction, and cerebrovascular accident; and (7) comorbidities or mental health abnormalities that could affect patients' participation in the study. Patients were eliminated from the study if: (1) radical gastrectomy could not be completed; (2) end-to-side esophagojejunal anastomosis was not performed during the procedure; or (3) esophagojejunal anastomosis reinforcement was not possible. Double and a half layered esophagojejunal anastomosis was performed as follows: (1) Open surgery: the full thickness of the anastomosis is continuously sutured, followed by embedding the seromuscular layer with barbed or 3-0 absorbable sutures. The anastomosis is sutured with an average of six to eight stitches. (2) Laparoscopic surgery: the anastomosis is strengthened by counterclockwise full-layer sutures. Once the anastomosis has been sutured to the right posterior aspect of the anastomosis, the jejunum stump is pulled to the right and the anastomosis turned over to continue to complete reinforcement of the posterior wall. The suture interval is approximately 5 mm. After completing the full-thickness suture, the anastomosis is embedded in the seromuscular layer. Relevant data of patients who had undergone radical gastrectomy in the above 12 centers from June 2021 were collected and analyzed. The primary outcome was safety (e.g., postoperative complications, and treatment). Other studied variables included details of surgery (e.g., surgery time, intraoperative bleeding), postoperative recovery (postoperative time to passing flatus and oral intake, length of hospital stay), and follow-up conditions (quality of life as assessed by Visick scores).Result:[1] From June 2021 to September 2022,457 patients were enrolled, including 355 men and 102 women of median age 60.8±10.1 years and BMI 23.7±3.2 kg/m2. The tumors were located in the upper stomach in 294 patients, mid stomach in 139; and lower stomach in 24. The surgical procedures comprised 48 proximal gastrectomies and 409 total gastrectomies. Neoadjuvant chemotherapy was administered to 85 patients. Other organs were resected in 85 patients. The maximum tumor diameter was 4.3±2.2 cm, number of excised lymph nodes 28.3±15.2, and number of positive lymph nodes five (range one to four. As to pathological stage,83 patients had Stage I disease, 128 Stage II, 237 Stage III, and nine Stage IV. [2] The studied surgery-related variables were as follows: The operation was successfully completed in all patients, 352 via a transabdominal approach, 25 via a transhiatus approach, and 80 via a transthoracoabdominal approach. The whole procedure was performed laparoscopically in 53 patients (11.6%), 189 (41.4%) underwent laparoscopic-assisted surgery, and 215 (47.0%) underwent open surgery. The median intraoperative blood loss was 200 (range, 10–1 350) mL, and the operating time 215.6±66.7 minutes. The anastomotic reinforcement time was 2 (7.3±3.9) minutes for laparoscopic-assisted surgery, 17.6±1.7 minutes for total laparoscopy, and 6.0±1.2 minutes for open surgery. [3] The studied postoperative variables were as follows: The median time to postoperative passage of flatus was 3.1±1.1 days and the postoperative gastrointestinal angiography time 6 (range, 4–13) days. The median time to postoperative oral intake was 7 (range, 2–14) days, and the postoperative hospitalization time 15.8±6.7 days. [4] The safety-related variables were as follows: In total, there were 184 (40.3%) postoperative complications. These comprised esophagojejunal anastomosis complications in 10 patients (2.2%), four (0.9%) being anastomotic leakage (including two cases of subclinical leakage and two of clinical leakage; all resolved with conservative treatment); and six patients (1.3%) with anastomotic stenosis (two who underwent endoscopic balloon dilation 21 and 46 days after surgery, the others improved after a change in diet). There was no anastomotic bleeding. Non-anastomotic complications occurred in 174 patients (38.1%). All patients attended for follow-up at least once, the median follow-up time being 10 (3–18) months. Visick grades were as follows: Class I, 89.1% (407/457); Class II, 7.9% (36/457); Class III, 2.6% (12/457); and Class IV 0.4% (2/457).Conclusion:Double and a half layered esophagojejunal anastomosis in radical gastrectomy is safe and feasible.

Objective:To evaluate the safety of double and a half layered esophagojejunal anastomosis in radical gastrectomy.Methods:This prospective, multi-center, single-arm study was initiated by the Affiliated Cancer Hospital of Zhengzhou University in June 2021 (CRAFT Study, NCT05282563). Participating institutions included Nanyang Central Hospital, Zhumadian Central Hospital, Luoyang Central Hospital, First Affiliated Hospital of Henan Polytechnic University, First Affiliated Hospital of Henan University, Luohe Central Hospital, the People's Hospital of Hebi, First People's Hospital of Shangqiu, Anyang Tumor Hospital, First People's Hospital of Pingdingshan, and Zhengzhou Central Hospital Affiliated to Zhengzhou University. Inclusion criteria were as follows: (1) gastric adenocarcinoma confirmed by preoperative gastroscopy;(2) preoperative imaging assessment indicated that R0 resection was feasible; (3) preoperative assessment showed no contraindications to surgery;(4) esophagojejunostomy planned during the procedure; (5) patients volunteered to participate in this study and gave their written informed consent; (6) ECOG score 0–1; and (7) ASA score I–III. Exclusion criteria were as follows: (1) history of upper abdominal surgery (except laparoscopic cholecystectomy);(2) history of gastric surgery (except endoscopic submucosal dissection and endoscopic mucosal resection); (3) pregnancy or lactation;(4) emergency surgery for gastric cancer-related complications (perforation, hemorrhage, obstruction); (5) other malignant tumors within 5 years or coexisting malignant tumors;(6) arterial embolism within 6 months, such as angina pectoris, myocardial infarction, and cerebrovascular accident; and (7) comorbidities or mental health abnormalities that could affect patients' participation in the study. Patients were eliminated from the study if: (1) radical gastrectomy could not be completed; (2) end-to-side esophagojejunal anastomosis was not performed during the procedure; or (3) esophagojejunal anastomosis reinforcement was not possible. Double and a half layered esophagojejunal anastomosis was performed as follows: (1) Open surgery: the full thickness of the anastomosis is continuously sutured, followed by embedding the seromuscular layer with barbed or 3-0 absorbable sutures. The anastomosis is sutured with an average of six to eight stitches. (2) Laparoscopic surgery: the anastomosis is strengthened by counterclockwise full-layer sutures. Once the anastomosis has been sutured to the right posterior aspect of the anastomosis, the jejunum stump is pulled to the right and the anastomosis turned over to continue to complete reinforcement of the posterior wall. The suture interval is approximately 5 mm. After completing the full-thickness suture, the anastomosis is embedded in the seromuscular layer. Relevant data of patients who had undergone radical gastrectomy in the above 12 centers from June 2021 were collected and analyzed. The primary outcome was safety (e.g., postoperative complications, and treatment). Other studied variables included details of surgery (e.g., surgery time, intraoperative bleeding), postoperative recovery (postoperative time to passing flatus and oral intake, length of hospital stay), and follow-up conditions (quality of life as assessed by Visick scores).Result:[1] From June 2021 to September 2022,457 patients were enrolled, including 355 men and 102 women of median age 60.8±10.1 years and BMI 23.7±3.2 kg/m2. The tumors were located in the upper stomach in 294 patients, mid stomach in 139; and lower stomach in 24. The surgical procedures comprised 48 proximal gastrectomies and 409 total gastrectomies. Neoadjuvant chemotherapy was administered to 85 patients. Other organs were resected in 85 patients. The maximum tumor diameter was 4.3±2.2 cm, number of excised lymph nodes 28.3±15.2, and number of positive lymph nodes five (range one to four. As to pathological stage,83 patients had Stage I disease, 128 Stage II, 237 Stage III, and nine Stage IV. [2] The studied surgery-related variables were as follows: The operation was successfully completed in all patients, 352 via a transabdominal approach, 25 via a transhiatus approach, and 80 via a transthoracoabdominal approach. The whole procedure was performed laparoscopically in 53 patients (11.6%), 189 (41.4%) underwent laparoscopic-assisted surgery, and 215 (47.0%) underwent open surgery. The median intraoperative blood loss was 200 (range, 10–1 350) mL, and the operating time 215.6±66.7 minutes. The anastomotic reinforcement time was 2 (7.3±3.9) minutes for laparoscopic-assisted surgery, 17.6±1.7 minutes for total laparoscopy, and 6.0±1.2 minutes for open surgery. [3] The studied postoperative variables were as follows: The median time to postoperative passage of flatus was 3.1±1.1 days and the postoperative gastrointestinal angiography time 6 (range, 4–13) days. The median time to postoperative oral intake was 7 (range, 2–14) days, and the postoperative hospitalization time 15.8±6.7 days. [4] The safety-related variables were as follows: In total, there were 184 (40.3%) postoperative complications. These comprised esophagojejunal anastomosis complications in 10 patients (2.2%), four (0.9%) being anastomotic leakage (including two cases of subclinical leakage and two of clinical leakage; all resolved with conservative treatment); and six patients (1.3%) with anastomotic stenosis (two who underwent endoscopic balloon dilation 21 and 46 days after surgery, the others improved after a change in diet). There was no anastomotic bleeding. Non-anastomotic complications occurred in 174 patients (38.1%). All patients attended for follow-up at least once, the median follow-up time being 10 (3–18) months. Visick grades were as follows: Class I, 89.1% (407/457); Class II, 7.9% (36/457); Class III, 2.6% (12/457); and Class IV 0.4% (2/457).Conclusion:Double and a half layered esophagojejunal anastomosis in radical gastrectomy is safe and feasible.

Objective:To evaluate the effects of objective feedback teaching applied in the specialist nurses training of digestive endoscopy.Methods:Six-eighty nurses participating in the training of specialist nurses of digestive endoscopy in Chongqing were recruited in the study, among which, 30 trainees in 2019 were assigned to the observation group, taking objective feedback teaching, and 38 trainees in 2018 were assigned to the control group, receiving traditional teaching. After the training, the two teaching methods were assessed through theoretical results, operation skills and comprehensive evaluation, and meanwhile a self-designed satisfaction questionnaire on the satisfaction of trainees was conducted. SPSS 25.0 was used to perform t-test, Chi-square test and Mann-Whitney U test. Results:The operation skills (84.90±4.92) and comprehensive quality (86.30±4.62) of observation group were higher than those of the control group [(82.39±4.10), (83.86±5.10)], with statistically significant differences ( P<0.05), while there was no significant difference in theoretical knowledge [(85.80±5.63) vs. (83.68±4.51), P>0.05]. Trainees were satisfied with the new training mode. Conclusion:The objective feedback teaching mode can improve the training effect especially on operation skills and comprehensive quality of the specialist nurses of digestive endoscopy, and insure the satisfaction of specialist nurses with the training.

Objective To investigate the CT features of clear cell renal carcinoma (ccRCC)and angiomyolipoma with minimal fat (AMLmf)and to improve the CT diagnostic accuracy of these two diseases.Methods The CT features of 55 patients with pathologically-confirmed ccRCC and 1 2 patients with pathologically-confirmed AMLmf were analyzed retrospectively,including the CT value in both plain and tri-phase enhanced CT scan,tumor enhancement rate(△R1,△R2,△R3),maximum diameter,enhanced homogeneity,location of the main tumor,cortex raising signs,etc.The statistical analysis was carried on.Results The maximum diameter,the CT value in parenchymal phase,enhancement rate (△R1,△R2,△R3)of tumors in ccRCC group were significantly higher than those of tumors in AMLmf group,and the CT value in plain CT scan in ccRCC group was significantly lower than that in AMLmf group (all P<0.05).No statistically significant difference was found in the CT value of the tumor in corticomedullary phase and in excretion phase (both P>0.05).The rate of extrarenally-located main tumors of AMLmf group was significantly higher than that of ccRCC group (P=0.020),the location of main tumors and cortex raising signs showed no statistically significant difference with the maximum tumor diameter(both P>0.05).The enhanced homogeneity of the tumor in corticomedullary phase,parenchymal phase and excretion phase in ccRCC group was lower than that in AMLmf group (all P<0.05).Conclusion The CT value in plain CT scan in ccRCC group is lower than that in AMLmf group;the enhancement rate of the ccRCC group is higher than that of the AMLmf group;the enhanced homogeneity of the ccRCC group is worse than that of the AMLmf group.The extrarenally-located main tumors are more commonly seen in AMLmf than in ccRCC,and the cortex raising signs and the location of main tumors are unrelated to the size of the tumor.

Objective To clarify the characteristics and postoperative benefit of gallbladder carcinoma in elderly patients (≥ 65 years old).Methods Two hundred and seventy-three patients of gallbladder carcinoma were collected,who were treated intent resection from January 2004 to December 2012 in the Department of Hepatobiliary Surgery,Eastern Hepatobiliary Hospital,Second Military Medical University.More than 65 years old was defined as the elderly,else was defined as the younger.The clinical-pathological features and prognosis of 85 elderly patients(elderly group) and 188 younger patients (younger group) were retrospectively analyzed.The survival of patients were followed up by telephone or outpatient.The incidence of hypertension,incidence of diabetes,TNM staging,and median CA19-9 were compared between the two groups.Continuous variables using a two-sample t test or Mann-Whitney U test.Categorical variables were compared by the Chi-square test or Fisher probability method.The survival curve was drawn by the Kaplan-Meier method.The univariate analysis and multivariate analysis of prognosis were respectively done using the Log-rank test and COX regression model.Results The incidence of hypertension,incidence of diabetes,TNM stage ratio (Ⅲ + Ⅳ/Ⅰ + Ⅱ),and CA19-9 median in the elderly group were 30.6％,11.8％,27.6 and 69.3 U/ml,respectively.The differences in the younger group were 13.8％,4.8％,7.9 and 28.2 U/ml,respectively,with statistically significant difference between the two groups (all P ＜ 0.05).The incidence of complications was 54.1％ in the elderly group and 48.9％ in the younger group,with no significant difference between the two groups (P =0.302).The median survival of the elderly group was 28.01 months,and the median survival of the younger group was 36.20 months,with no statistical difference between the two groups (P =0.131).Cox analysis showed that independent prognostic risk factors for the elderly patients with gallbladder cancer included liver invasion (HR =2.386,95％ CI:1.379-4.127,P =0.002) and lymph node metastasis (HR =1.866,95 ％ CI:1.100-3.167,P =0.021).Conclusions Radical resection is safe and feasible for elderly patients with gallbladder carcinoma.Age is not a contraindication for surgery.Radical resection can get the same benefits as young people.Liver invasion and lymph node metastasis are independent risk factors affecting the prognosis of the elderly patients with gallbladder carcinoma.

Objective To investigate the clinical significance of joint application of carcinoembryionic and carbohydrate antigen 19-9 for improving the prognosis of patients with gallbladder cancer.Methods A retrospective analysis of clinical-pathological features and prognosis were conducted including 390 patients with gallbladder cancer,who were treated from January 2003 to December 2013 at Eastern Hepatobiliary Surgery Hospital of Second Military Medical University.The clinical value of joint application of careinoembryionic and carbohydrate antigen 19-9 in prognosis was explored.The survival of patients was followed up by telephone or outpatient.Continuous variables were indicated as median (average) and categorical variables were expressed as number(n).Mann-Whitney U test was applied for continuous variables.Categorical variables were compared by the Chi-square test or Fisher probability method.The survival curve was drawn by the Kaplan-Meier method.The univariate analysis and multivariate analysis of prognosis were respectively done using the Log-rank test and COX regression model.Results Of the 390 patients,327 had complete data.The median (range) follow-up time was 49 (1-123) months.Both carcinoembryionic and carbohydrate antigen 19-9 could be used to predict prognosis of gallbladder cancer.A linear combination of carcinoembryionic and carbohydrate antigen 19-9 was significantly better(0.9365) than carbohydrate antigen 19-9 (0.7619) as well as carcinoembryionic (0.7937) alone in term of specificity.Preoperative blood test carcinoembryionic and carbohydrate antigen 19-9 nature showed double-negative group had the best prognosis,with a median survival of 27.06 months,R0 radical rate was 86.1％.Doublepositive group had the worst prognosis,with a median survival of 6.17 months,R0 radical rate was 45.1％.Conclusions Combinations of pretherapeutic tumor makers carcinoembryionic and carbohydrate antigen 19-9 can improve the accuracy of predicting prognostic in patients with gallbladder cancer,and the clinical application is practical.

BACKGROUND:Rabbit bone marrow mesenchymal stem cels as a kind of adult stem cels with strong proliferation and multilineage differentiation potential exhibit a tremendous application potential in tissue engineering and biological therapy.
OBJECTIVE:To in vitro culture, proliferate and identify rabbit bone marrow mesenchymal stem cels and to observe cel biological characteristics.
MEHTODS:Bone marrow of rabbits was extracted under sterile conditions to separate bone marrow mesenchymal stem cels using the whole bone marrow adherence method and Percol density gradient centrifugation method. Afterwards, the cels were purified and proliferated using differential adherence method. Morphology and growth pattern of cels were observed under microscope, and expression of cel surface antigen markers was detected by flow cytometry.
RESULTS AND CONCLUSION:Rabbit bone marrow mesenchymal stem cels presented with short adherent time and fast growth. After passage and purification, impurities cel counts were decreased. Primary cels presented with triangular, fusiform and spindly shapes. Passage 5 cels with single shape showed the typical polar swirling growth, and could not express CD34 and CD45, but expressed CD29 and CD44. These findings indicate that the cels cultured using the whole bone marrow adherence method and Percol density gradient centrifugation method possess stem cel characteristics in morphology, surface markers and multilineage differentiation, which have been identified as bone marrow mesenchymal stem cels by flow cytometry.

Objective To review the pathogenesis of osteoarthritis ( OA) and microsphere delivery systems advances in the field of osteoarthritis treatment, and provide theoretical support for clinical research and osteoarthritis therapy.Methods The literatures on osteoarthritis have been reviewed in recent years, and pathogenesis and osteoarthritis therapy were summarized.ResuIts At present, the therapy of OA was gradually from symptoms relief to pathogenesis, and further revealed the inner regularity of osteoarthritis treatment.However, a variety of western medicines had its own limitations. ConcIusion In recent years, intra-articular injection therapy because of its unique advantages in clinical treatments,attracts widespread attention, and particularly the microsphere injection with superior prospects is researched more in the field of new formulations.

Objective To study optimal compatibility scheme of vidarabine monophosphate for injection combined with aciclovir injection. Methods L18(35) of orthogonal design was used with five factors: temperature, illumination,storage time,solvent and solvent dosage;the insoluble particles were observed by GWJ-4 type particle analyzer.The content of vidarabine monophosphate and aciclovir was detected by HPLC.Optimization of the best scheme of vidarabine monophosphate for Injection combined with aciclovir injection was studied.Results The regression equation of vidarabine monophosphate and aciclovir were A=3.78 ×104C+2.32, r=0.9997(n=5)and A=7.11 ×105C-4.65, r=0.9995(n=5),The results showed that the relationship between the range of 200-900μg/mL(vidarabine monophosphate) and the range of 1000-4500μg/mL(aciclovir) was good.The optimal compatibility conditions are the follows:temperature of 25,as far as possible away from light;Vidarabine monophosphate for injection 1 (0.1 g/branch) and 1 acyclovir injection (10 mL:0.5 g) mixed with 100 mL 0.9% sodium chloride injection,and dropping out within 4 h after compatibility. Conclusion The optimal compatibility scheme has good repeatability,injection solution stability is good.It prouides scientific reference for the safty of clinical rational drug use.