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Traditional chemotherapy is the cornerstone of comprehensive treatment for gastric cancer, but its recurrence and metastasis rates are high, and the overall prognosis is not ideal. The rise of immune checkpoint inhibitors (ICI) has brought new hope to gastric cancer patients and changed the current pattern of comprehensive treatment for gastric cancer. With the increasing use of ICI, immune related adverse reactions (irAEs) such as skin toxicity and gastrointestinal toxicity are becoming increasingly common. Scientific understanding, early diagnosis, and graded management are currently the main strategies for handling irAEs. This article aims to review the mechanisms, clinical manifestations, and prediction, treatment, and management of irAEs after ICI treatment of gastric cancer, in order to enhance the understanding of irAEs among clinical physicians, better manage immunotherapy related adverse reactions, and improve the prognosis and quality of life of patients.
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Objective:To investigate the prognosis of patients with initially resectable gastric cancer liver metastasis (GCLM) who were treated by different modalities, and analyze the influencing factors for prognosis of patients.Methods:The retrospective cohort study was conducted. The clinicopathological data of 327 patients with initially resectable GCLM who were included in the database of a nationwide multicenter retrospective cohort study on GCLM based on real-world data from January 2010 to December 2019 were collected. There were 267 males and 60 females, aged 61(54,68)years. According to the specific situations of patients, treatment modalities included radical surgery combined with systemic treatment, palliative surgery combined with systemic treatment, and systemic treatment alone. Observation indicators: (1) clinical characteristics of patients who were treated by different modalities; (2) prognostic outcomes of patients who were treated by different modalities; (3) analysis of influencing factors for prognosis of patients with initially resectable GCLM; (4) screening of potential beneficiaries in patients who were treated by radical surgery plus systemic treatment and patients who were treated by palliative surgery plus systemic treatment. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. The Kaplan-Meier method was used to calculate survival rate and draw survival curve, and Log-Rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the COX proportional hazard regression model. The propensity score matching was employed by the 1:1 nearest neighbor matching method with a caliper value of 0.1. The forest plots were utilized to evaluate potential benefits of diverse surgical combined with systemic treatments within the population. Results:(1) Clinical characteristics of patients who were treated by different modalities. Of 327 patients, there were 118 cases undergoing radical surgery plus systemic treatment, 164 cases undergoing palliative surgery plus systemic treatment, and 45 cases undergoing systemic treatment alone. There were significant differences in smoking, drinking, site of primary gastric tumor, diameter of primary gastric tumor, site of liver metastasis, and metastatic interval among the three groups of patients ( P<0.05). (2) Prognostic outcomes of patients who were treated by different modalities. The median overall survival time of the 327 pati-ents was 19.9 months (95% confidence interval as 14.9-24.9 months), with 1-, 3-year overall survival rate of 61.3%, 32.7%, respectively. The 1-year overall survival rates of patients undergoing radical surgery plus systemic treatment, palliative surgery plus systemic treatment and systemic treatment alone were 68.3%, 63.1%, 30.6%, and the 3-year overall survival rates were 41.1%, 29.9%, 11.9%, showing a significant difference in overall survival rate among the three groups of patients ( χ2=19.46, P<0.05). Results of further analysis showed that there was a significant difference in overall survival rate between patients undergoing radical surgery plus systemic treatment and patients undergoing systemic treatment alone ( hazard ratio=0.40, 95% confidence interval as 0.26-0.61, P<0.05), between patients undergoing palliative surgery plus systemic treatment and patients under-going systemic treatment alone ( hazard ratio=0.47, 95% confidence interval as 0.32-0.71, P<0.05). (3) Analysis of influencing factors for prognosis of patients with initially resectable GCLM. Results of multivariate analysis showed that the larger primary gastric tumor, poorly differentiated tumor, larger liver metastasis, multiple hepatic metastases were independent risk factors for prognosis of patients with initially resectable GCLM ( hazard ratio=1.20, 1.70, 1.20, 2.06, 95% confidence interval as 1.14-1.27, 1.25-2.31, 1.04-1.42, 1.45-2.92, P<0.05) and immunotherapy or targeted therapy, the treatment modality of radical or palliative surgery plus systemic therapy were independent protective factors for prognosis of patients with initially resectable GCLM ( hazard ratio=0.60, 0.39, 0.46, 95% confidence interval as 0.42-0.87, 0.25-0.60, 0.30-0.70, P<0.05). (4) Screening of potentinal beneficiaries in patients who were treated by radical surgery plus systemic treatment and patients who were treated by palliative surgery plus systemic treatment. Results of forest plots analysis showed that for patients with high-moderate differentiated GCLM and patients with liver metastasis located in the left liver, the overall survival rate of patients undergoing radical surgery plus systemic treatment was better than patients undergoing palliative surgery plus systemic treatment ( hazard ratio=0.21, 0.42, 95% confidence interval as 0.09-0.48, 0.23-0.78, P<0.05). Conclusions:Compared to systemic therapy alone, both radical and palliative surgery plus systemic therapy can improve the pro-gnosis of patients with initially resectable GCLM. The larger primary gastric tumor, poorly differen-tiated tumor, larger liver metastasis, multiple hepatic metastases are independent risk factors for prognosis of patients with initial resectable GCLM and immunotherapy or targeted therapy, the treatment modality of radical or palliative surgery plus systemic therapy are independent protective factors for prognosis of patients with initially resectable GCLM.
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Objective:To identify the risk factors associated with postoperative adjuvant chemotherapy in patients with stage I gastric cancer and establish nomograms model based on risk factors.Methods:In this retrospective case-control study, 161 cases with stage Ⅰ primary gastric adenocarcinoma were included who underwent gastrectomy at the Department of General Surgery of the First Medical Center of Chinese PLA General Hospital from January to December in 2020, including 129 male cases and 32 females cases, with the average age of (59.90±0.80) years. Among them, 41 cases were treated with postoperative adjuvant chemotherapy (chemotherapy group), while 120 cases who did not receive postoperative adjuvant chemotherapy (no chemotherapy group). Univariate and multivariate Logistic regression analyses were used to identify the risk factors of adjuvant chemotherapy in stage Ⅰ gastric cancer patients and establish the nomograms predictive model. ROC curve and calibration curve were used to evaluate the performance of the model.Results:Multivariate analysis revealed that primary tumor site, tumor size, T stage, N stage lymph-vascular tumor embolus or perineural invasion were the independent risk factors of postoperative adjuvant chemotherapy for stage Ⅰ gastric cancer( P<0.05). The ROC curve indicated that area under the curve (AUC) of the multivariate model was 0.91(95% CI: 0.86-0.97). The calibration curve showed that probability predicted by nomograms was consistent with the actual situation(C-index: 0.91). Conclusions:The tumor located in the proximal stomach, tumor size>2 cm, T 2, N 1, lymph-vascular tumor embolus or perineural invasion maybe be the risk factors for chemotherapy decision in stage Ⅰ gastric cancer patients. The established model has good predictive ability for postoperative chemotherapy of stage Ⅰ gastric cancer patients, which might provide reference for the selection of clinical decisions in this part of patients.
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Objective:To study the risk factors of complications after bowel resection for acute mesenteric ischemic disease.Methods:Retrospective case-control study was used to analyze the case data of 68 patients diagnosed with acute mesenteric ischemic disease (AMI) with bowel resection at the First Medical Center of the PLA General Hospital from January 2010 to January 2020, including 43 males and 25 females. The patients were divided into complication group ( n=21) and the non-complication group ( n=47) according to whether they had complications after surgery. The risk factors associated with the development of postoperative complications were analyzed by multivariate Logistic stepwise regression method to determine the risk factors with clinical significance. Measurement data with normal distribution were expressed as mean ± standard deviation ( ± s), and t-test was used for comparison between groups. Chi-square test was used for comparison between count data groups. Results:Univariate analysis showed that age >60 years, Marshall score≥2, type of resected bowel, pathology suggestive of irreversible transmural necrosis, length of ICU stay >6 d, length of mechanical ventilation >2 d, American Society of Anesthesiologists (ASA) classification, and preoperative procalcitonin≥2 ng/mL were the risk factors affecting the development of complications after bowel resection for acute mesenteric ischemic disease risk factors ( P<0.05). Multivariate Logistic regression analysis showed that age>60 years ( HR=12.364, 95% CI: 1.135-134.662, P=0.039) and preoperative procalcitonin ≥2 ng/mL ( HR=14.144, 95% CI: 1.280-156.303, P=0.031) were independent risk factors for the development of postoperative complications after AMI parallel bowel resection. Conclusion:The rate of complications after combined bowel resection for AMI is high. When patients are combined with age>60 years and preoperative procalcitonin≥2 ng/mL, preoperative prevention of postoperative complications should be emphasized to improve the prognosis of patients.
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Acute mesenteric ischemic disease(AMI) is a critical surgical emergency. Due to its insidious and atypical initial symptoms, a delayed diagnosis significantly increases the mortality rate of AMI patients. The diagnosis and treatment process of AMI often involve close collaboration and communication among multiple disciplines and departments.Our team deeply recognizes the importance of a multidisciplinary team(MDT) model in the standardized management of the AMI disease. It can significantly improve the efficiency of disease diagnosis, delay disease progression, provide timely treatment, and improve the prognosis of patients, thereby reducing the overall mortality rate. Therefore, combining our center′s experience in treating AMI patients with the implementation of the MDT model, we elaborated on the key steps and quality control measures for the standardized application of the MDT model in AMI disease, with the aim of providing a theoretical foundation and experiential reference for colleagues and professionals.
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Objective To analyze the occurrence and clinical significance of hypophosphatemia after mod-erate-to-major abdominal surgery. Methods A total of 120 patients who had received moderate to major abdominal surgery and then transferred to the intensive care unit ( ICU) from January 2008 to October 2018 were retrospectively analyzed. The peri-operative parameters of serum inorganic phosphate and calcium levels were recorded and analyzed. The incidence of hypophosphatemia after the operation was calculated. The effect of phosphorus-supplement infusion on the complications and mortality within 30 days after the operation were observed. Results After moderate-to-major surgery, the phosphorus concentration significantly decreased [ (1. 21±0. 27) vs. (0. 83±0. 24) mmol/L, P<0. 05];the incidence of hypophosphatemia was 58. 3%, especially in patients with nutritional risk assessed before the operation. The post-operative complications were less in the phosphorus-supplement infusion group than in the control group (17. 5%vs. 41. 3%, P=0. 028). Conclusion The incidence of hypophosphatemia is relatively high after moderate-to-major abdominal surgery, especially in patients with nutritional risks. The treatment of phosphorus-sup-plement infusion can benefit the patients in their short-term prognosis.
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Human epidermal growth factor receptor 2 (HER2) proteins are overexpressed in a high proportion of gastric cancer (GC) cases and affect the maintenance of cancer stem cell (CSC) subpopulations, which are used as targets for the clinical treatment of patients with HER2-positive GC. Despite improvements in survival, numerous HER2-positive patients fail treatment with trastuzumab, highlighting the need for more effective therapies. In this study, we generated a novel type of genetically modified human T cells, expressing a chimeric antigen receptor (CAR), and targeting the GC cell antigen HER2, which harbors the CD137 and CD3ζ moieties. Our findings show that the expanded CAR-T cells, expressing an increased central memory phenotype, were activated by the specific recognition of HER2 antigens in an MHC-independent manner, and effectively killed patient-derived HER2-positive GC cells. In HER2-positive xenograft tumors, CAR-T cells exhibited considerably enhanced tumor inhibition ability, long-term survival, and homing to targets, compared with those of non-transduced T cells. The sphere-forming ability and in vivo tumorigenicity of patient-derived gastric cancer stem-like cells, expressing HER2 and the CD44 protein, were also inhibited. Our results support the future development and clinical application of this adoptive immunotherapy in patients with HER2-positive advanced GC.
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Animals , Humans , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasms, Experimental , Allergy and Immunology , Pathology , Therapeutics , Receptor, ErbB-2 , Allergy and Immunology , Receptors, Antigen, T-Cell , Allergy and Immunology , Stomach Neoplasms , Allergy and Immunology , Pathology , Therapeutics , Tumor Cells, CulturedABSTRACT
Objective@#To compare the short-term and long-term outcome between robotic gastrectomy and laparoscopic gastrectomy.@*Methods@#The clinical data of 517 patients who had received robotic gastectomy and laparoscopic gastrectomy between December 2011 and December 2013 at Department of General Surgery, Chinese People′s Liberation Army General Hospital was collected. After propensity score matching, 70 patients in robotic gastectomy and 70 patients in laparoscopic gastectomy were identified. Perioperative outcome and overall survival were compared between the two groups using t test, χ2 test, Kaplan-Meier curve and Log-rank test, respectively. Prognosis factors were analyzed by Cox′s proportional hazards regression.@*Results@#There were comparable baseline characteristics between patients in robotic group (RG) and those in laparoscopic group (LG). The conversion rate for RG and LG were 5.7% and 4.3% respectively (P=1.000). Compared with LG, RG had similar lymph node retrieval (25.5±7.2 vs. 24.5±8.3, t=0.770, P=0.443) and less blood loss ((147.0±96.8) ml vs. (188.0±111.2) ml, t=-2.326, P=0.021). There were also similar complications (χ2=0.233, P=0.629) and severity of complications (W=70.500, P=0.053). Although there tended to be early mobility, early flatus and less hospital stay for patients in RG group, the difference between RG and LG was not statistically significant. The 3-year survival rate was 72.9% and 60.0% for patients in RG and patients in LG (P=0.578). Multivariable analysis revealed gender (HR=2.529, 95% CI: 1.042 to 6.140, P=0.040), neoadjuvant chemotherapy (HR=0.272, 95% CI: 0.104 to 0.710, P=0.008) and vascular invasion (HR=2.135, 95% CI: 1.027 to 4.438, P=0.042) were independent prognostic factors.@*Conclusion@#Compared with laparoscopic gastrectomy, robotic gastectomy could achieve similar short-term and long-term outcomes.
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<p><b>OBJECTIVE</b>To compare the short-term outcomes between robotic and laparoscopic radical total gastrectomy in gastric cancer patients with BMI index ≥24 kg/m.</p><p><b>METHOD</b>Clinical data of 93 gastric cancer patients who underwent robotic and laparoscopic radical total gastrectomy at PLA General Hospital from April 2016 to April 2017 were retrospectively analyzed. The retrospective cohort study was adopted.</p><p><b>INCLUSION CRITERIA</b>preoperatively definite diagnosis of primary gastric cancer by endoscopy and biopsy; preoperative BMI ≥24 kg/m; no previous abdominal surgery; no previous chemotherapy and radiotherapy; no distant metastasis or invasion into adjacent organs before operation or during operation; receiving radical gastrectomy; Roux-en-Y reconstruction of digestive tract in open procedure. According to approaches of minimally invasive surgery, 24 patients underwent robotic surgery and 69 underwent laparoscopic surgery. The intraoperative parameters (overall operative time, pneumoperitoneal time, open procedure time, intraoperative blood loss, transfusion rate, number of total retrieved lymph nodes and metastatic lymph nodes) and postoperative parameters (drainage in the first postoperative day, the first defecation time, morbidity of postoperative complication and hospital stay) were compared between two groups. Correlation of the above parameters were analyzed.</p><p><b>RESULTS</b>Of 93 patients, 77 were male and 16 female with an average age of (60.0±10.6) years. The average BMI was (26.8±1.3) kg/m in whole patients, (26.9±1.6) kg/m in robotic group and (26.8±1.7) kg/m in laparoscopic group. No significant differences in age, gender, BMI, preoperative ASA class, postoperative pathological findings and clinical classification were observed between two groups, which made short-term parameters between two groups comparable. The robotic group had a significantly longer overall operative time [(301.2±68.9) minutes vs. (247.3±59.6) minutes, P=0.000], longer open procedure time [(141.5±26.3) minutes vs. (92.5±36.7) minutes, P=0.029] and higher cost than laparoscopy group[(17.5×10 ± 9.7×10) yuan vs. (10.0×10 ± 2.3×10) yuan, P=0.001]. Pneumoperitoneal operative time, intraoperative blood loss, transfusion rate, number of total retrieved lymph nodes, number of harvested metastatic lymph nodes and postoperative short-term efficacy were similar between the two groups (all P>0.05). In robotic group, pneumoperitoneal operative time was positively correlated with overall operative time (r=0.708, P=0.010); total cost was positively correlated with postoperative hospital stay (r=0.493, P=0.000) and open procedure time was negatively correlated with the first defecation time (r=-0.962, P=0.038). In laparoscopy group, total cost was positively correlated with overall operative time (r=0.411, P=0.046), drainage volume in the first postoperative day was positively correlated with the number of total dissected lymph node (r=0.540, P=0.006), postoperative hospital stay was positively correlated with intraoperative blood loss (r=0.574, P=0.003), total cost was positively correlated with intraoperative blood loss and hospital stay (r=0.609, P=0.002; r=0.865, P=0.000), drainage volume in the first postoperative day was positively correlated with BMI (r=0.533, P=0.007).</p><p><b>CONCLUSION</b>For gastric cancer patients with BMI ≥24 kg/m, robotic radical total gastrectomy is associated with longer operative time and higher cost, but is less vulnerable to the change of BMI and more in favor of the realization of enhanced recovery after surgery (ERAS) than laparoscopic radical total gastectomy.</p>
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Aged , Female , Humans , Male , Middle Aged , Body Mass Index , Gastrectomy , Methods , Laparoscopy , Length of Stay , Operative Time , Retrospective Studies , Robotic Surgical Procedures , Stomach Neoplasms , General Surgery , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinicopathologic features and prognostic factors of gastric neuroendocrine neoplasms(gNENs).</p><p><b>METHODS</b>Clinicopathologic data of 104 patients with gastric neuroendocrine neoplasms admitted in Chinese PLA General Hospital between January 2000 and December 2014 were analyzed retrospectively. Tumor proliferation activity classification (G1, G2 and G3) and TNM staging were observed. The clinicopathologic features of the whole group were collected and the univariate and multivariate analysis were determined by Log-rank and Cox proportional hazard model to detect the prognosis-determining features.</p><p><b>RESULTS</b>Of all the patients, 66 cases(63.5%) were neuroendocrine carcinoma, 25 cases(24.0%) were mixed adenoendocrine carcinoma and 12 cases (11.5%) were neuroendocrine tumor. For G grades, 92 cases (88.5%) were G3 grade, 8 cases(7.7%) were G2 grade and 4 cases (3.8%) were G1 grade. TNM staging results showed that stageI( was found in 6 cases (5.8%), stageII(A in 6 cases (5.8%), stageII(B in 9 cases (8.7%), stage III(A in 8 cases (7.7%), stage III(B in 55 cases (52.9%) and stageIIII( in 20 cases (19.2%). For T stage, 7 cases (6.7%) were T1, 12 cases (11.5%) were T2, 24 cases (23.1%) were T3, and 61 cases (58.7%) were T4. Lymph node metastasis occurred in 73 cases (70.2%) and distant metastasis occurred in 20 cases(19.2%). Eighty-six patients were followed up for 6 to 186 months. The median survival was 33.0 months(95% CI: 28.3 to 36.6), and 1-, 3-, and 5-year survival rates were 80%, 49% and 31%. Clinicopathologic features which were considered statistically significant on univariate analysis were selected to Cox proportional hazard model. Univariate analysis showed that risk factors of reducing survival rate included tumor size, pathological type, proliferation activity grades, and depth of invasion (all P<0.05), as well as chromogranin A expression, tumor staging, lymph node metastasis and distant metastasis(all P<0.01). The multivariate analysis showed that the stage of gNEN was the independent risk factor of the prognosis (RR=14.213, 95% CI: 1.316 to 153.524, P=0.029).</p><p><b>CONCLUSION</b>Late staging is the main clinical feature and a prognostic factor for gNENs.</p>
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Humans , Carcinoma , Diagnosis , Pathology , Lymphatic Metastasis , Multivariate Analysis , Neoplasm Staging , Neuroendocrine Tumors , Diagnosis , Pathology , Prognosis , Proportional Hazards Models , Retrospective Studies , Risk Factors , Stomach Neoplasms , Diagnosis , Pathology , Survival RateABSTRACT
<p><b>OBJECTIVE</b>To systemically evaluate the relationship between the expression of insulin-like growth factor receptor type I (IGF-1R) and prognosis in gastric cancer (GC) patients.</p><p><b>METHODS</b>A literature search was conducted from PubMed, EMBASE, Web of Science, CNKI, Wanfang and VIP databases to retrieve the clinical studies relevant to IGF-1R expression and its prognostic value in GC patients. Meta-analysis was performed using STATA 12.0 software. The methodology was assessed according to the European Lung Cancer Working Party Quality Scale for Biological Prognostic Factors for Lung Cancer. The quality of studies was assessed using the Newcastle-Ottawa scale.</p><p><b>RESULTS</b>Four eligible studies including 685 patients were enrolled for this meta-analysis. Analysis results suggested that up-regulation of IGF-1R in GC patients was significantly associated with TNM staging (OR=5.20, 95%CI:1.12 to 24.15, P=0.035), lymph node metastasis(OR=8.24, 95%CI:2.68 to 25.34, P=0.000) and distant metastasis(OR=17.34, 95%CI:6.52 to 46.15, P=0.000). Moreover, up-regulated IGF-1R expression was significantly associated with poor overall survival of gastric cancer patients(HR=2.63, 95% CI:1.29 to 5.40, Z=2.64, P=0.008).</p><p><b>CONCLUSION</b>High IGF-1R expression may be an adverse prognostic factor in gastric cancer patients.</p>
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This paper is aimed to isolate and to cultivate human adipose-derived stem cells (hADSCs) from the adipose tissue by a combination of collagenase digestion, adherence to flasks and monoclonal cultural method so as to observe the morphological characteristics of the hADSCs. The immunophenotypes of hADSCs were detected by flow cytometry techniques. The general morphological characteristics of hADSCs were observed by cytochemical and immunofluorescent techniques. The ultrastructure of hADSCs was observed by transmission electron microscopy (TEM). The experimental results showed that hADSCs had unique immunophenotypes and they were positive for CD29, CD44, CD90, CD105 and CD166, but negative for CD31, CD45 and HLA-DR. Cytochemistry showed that cytoplasm of hADSCs was stained with light blue by hematoxylin-eosin, negative for Oil red O and AKP, and positive for immunofluorescence CD29 and CD166. There were abundant organella and microvilli in the ultrastructure of hADSCs. The results validate that they will offer a morphological foundation for application of the hADSCs.
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Humans , Adipose Tissue , Cell Biology , Cell Culture Techniques , Methods , Cells, Cultured , Microscopy, Electron, Transmission , Stem Cells , Cell Biology , Tissue Engineering , MethodsABSTRACT
BACKGROUND:Up to date,there is not an acceptable method for isolating,culture and amplifying human adipose-derived mesenchymal stem cells (hADSCs).OBJECTIVE:To explore the most effective way to obtain highly homogenous and undifferentiated hADSCs.DESIGN,TIME AND SETTING:The in vitro cytology experiment was performed at the Key Laboratory of Pathobiology,Ministry of Education,Jilin University from June to December 2008.MATERIALS:Human abdominal adipose tissue resected in the surgery was supplied by the Affiliated Hospital of Jilin University,The informed consent was obtained from patients.METHODS:Human adipose tissue was removed connective tissue and blood vessel,followed by incubation in 0.1% type I collagenase for 60 minutes at 37℃,filtrated then centrifuged.Consequently,the subnatant precipitation was cultured with LG-DMEM containing 10% fetal bovine serum,incubated at 6-well plate with density of 1×10~9/L,and placed in incubator of 5% CO_2 at 37 ℃.The cultured cells were passaged when the cells reached 80%-90% confiuency,and the 3~(rd) passage of cells were induced to osteogenic and adipogenic differentiation.MAIN OUTCOME MEASURES:Morphological characteristics of hADSCs were observed by laser scanning confocal microscope.Immunophenotypes,cell cycle and growth curve of hADSCs were detected by flow cytometry and immunofiuorescent techniques.In addition,the multiple differentiation potential of hADSCs was detected.RESULTS:hADSCs presented fibroblast-like morphological feature with a flocked array.The 3~(rd) passage of hADSCs had unique immunophenotypes and they were positive for CD73,CD44,CD166,CD105 and CD29,but negative for CD31,CD34,CD45 and HLA-DR.Cell cycle result showed that they had the typical growth characteristics of stem calls,namely,83.81% cells stayed at G_0/G_1 stage,only 16.19% cells were stayed at S+G_2/M stage;The latent phase of the primary culture cells was 2 days prior to and after incubation,followed by 3-6 days of logarithmical proliferation,reached a peak at day 6,and entering the growth platform phase with lower growth speed.The alkaline phosphatase was positive expressed at week 2 of osteogenic induction.And the positive expression of oil-O red staining could be seen at day 3 of adipogenic induction.CONCLUSION:Cells contamination can be reduced by removing connective tissue and blood vessel of adipose tissue,and 0.1% type Ⅰ collagenase for 60 minutes can effective separated stroma cell to matrix fiber,furthermore,ensure a sufficient contact between collagenase and tissures,which provide an supportive for harvesting highly homogenous hADSCs.
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Objective To investigate the TH1/TH2 patterns by calculating the percentage of TH1, TH2 cells, and the TH1/TH2 ratio of peripheral blood from patients with pregnancy-induced hypertension (PIH). Methods The percentage of TH1 and TH2 cells in peripheral blood from 15 normal pregnant controls and 20 PIH patients (including 12 moderate and 8 severe cases) were calculated using flow cytometry for the analysis of both the surface marker CD3 CD8 and intracellular cytokines, interleukin-4 (IL-4), and interferon-? (IFN-?). Results The percentage of TH1 cells and the ratio of TH1/TH2 in PIH patients were significantly higher than those in the normal third-trimester pregnant controls [(20.50?4.02)% vs (12.57?2.18)%, P