Objective To explore the effect of compound sophora flavescens injection on intraperitoneal chemotherapy of cisplatin in malignant ascites of gastric cancer patients and its influence on immune function.Methods 90 cases with malignant ascites of gastric cancer failures were researched,who from May 2014 to May 2016.According to random number table, they were divided into observation group and control group, 45 cases in each group.The control group was treated with intraperitoneal chemotherapy of cisplatin alone, and the treatment group was treated by intraperitoneal chemotherapy with compound Kushen injection combined with cisplatin.The two groups of patients before and after treatment, compared the level of immune function indicators, while observing the two groups of patients after treatment, short-term efficacy, quality of life improvement and adverse reactions.Results Observation group of patients with efficient and control rates were higher than the control group patients, but no obvious difference between groups.The stability factor of 55.55%of patients with observation group was obviously higher than that of control group 22.22%of the patients, the clear difference between groups (P<0.05).Before the treatment, two groups of patients of CD3 +, CD4 +, CD8 +and CD4 +/CD8 +T cell subsets and NK cells was no significant difference.After treatment, two groups of patients of CD3 +, CD4 +, CD4 +/CD8 + T cell subsets and NK cells were significantly increased than before treatment ( P<0.05 ) , and CD8 + than before treatment significantly reduced ( P <0.05 ) , while the observation group of CD3 +, CD4 +, CD4 +/CD8 +T cell subsets and NK cells was significantly higher than that of control group ( P <0.05 ) , and CD8 + was significantly lower than control group (P <0.05).The control group in the improvement of quality of life of patients with effective rate of 35.55%compared with 80.00% of the observation group of patients, significantly lower, more obvious differences between groups (P <0.05).Observation group of patients the incidence of gastrointestinal reaction and liver function abnormalities were 0.00%, 8.88%, significantly lower than the control group patients were 11.11%, 22.22%, there exists significant difference between groups (P <0.05).Fatigue, nausea, vomiting and 2 groups of patients compared the incidence rate of no significant difference.Conclusion The combined use of compound Kushen injection combined with cisplatin intraperitoneal perfusion chemotherapy in patients with malignant ascites of gastric cancer for treatment,effect is remarkable,Can effectively improve the patient's T cell subsets and NK cells,Improve the patient's immune function.

Objective To investigate the effect of vascular endothelial growth factor C (VEGF-C) on apoptosis of pancreatic cancer cell. Methods Human pancreatic cancer cell line PANC-1 orthotopic implantation tumor model was established in nude mice. Primary pancreatic cancer cells and that derived from lymphatic metastasis were primarily cultured. Expression of VEGF-C was inhibited through antisense oligodeoxynucleotide in vitro transfection. Reverse transcription polynlerase chain reaction (RT-PCR) and flow cytometer were used to detect the effect of VEGF-C on apoptosis of pancreatic cancer cells and bcl-2. Results After in vitro transfection, mRNA expression level of VEGF-C in PANC-1 pancreatic cancer cells significantly decreased (P ＜0. 01 ). Apoptosis rate of pancreatic cancer cells derived from spontaneous lymphatic metastasis was (2. 83 ± 1.01 ) %, ( 4. 98 ± 2. 05 ) %,and ( 13.22 ±2. 17) % respectively for control group, SODN group and ASODN group after in vitro transfection among which apoptosis rate in ASODN group increased significantly (P ＜0. 01 ). However, apoptosis rate for pancreatic cancer cells derived from primary tumor had no obvious change (P ＞0.05), with (3.51 ±1.38)%, (4.76 ±2. 16 ) %, and (5. 33 ± 2. 18 ) % respectively in control group, SODN group and ASODN group. The expression level of bcl-2 in pancreatic cancer cells derived from spontaneous lymphatic metastasis decreased significantly (P ＜0. 05) while it had no obvious change in primary pancreatic cancer cells (P ＞ 0. 05). Conclusion To inhibit expression of VEGF-C in pancreatic cancer cell can promote apoptosis of pancreatic cancer cell, which is relevant to downregulation of bcl-2;however, it has no obvious effect on primary pancreatic cancer.

Objective To study the effect of low molecular weight heparin in the treatment of hypercoagulable state of ulcerative colitis .Methods 116 patients with ulcerative colitis hypercoagulable were enrolled in the study . Thepatients were divided into two groups according to the random number table ,58 cases in each group .The conven-tional group was treated with conventional therapy ,and the low molecular weight heparin group was treated with low molecular weight heparin on the basis of conventional therapy .Both two groups were treated for 15 days.The degree of hypercoagulability ,the inflammatory status of the patients before and after treatment , and the platelet count and plasma D-dimer levels before and after treatment were compared .Results The improvement rate of hypercoagulable state of the low molecular weight heparin group was 96.55％,which was higher than 87.50％ in the routine group (χ2 =4.921,P<0.05).There were no statistically significant differences in platelet count and plasma D -dimer level between the two groups before treatment (all P<0.05).The platelet count and plasma D-dimer level were significantly lower in the low molecular weight heparin group(t=7.255,8.013,all P<0.05).The levels of TNF-αand CRP in the low molecular weight heparin group were significantly lower than those in the control group ( t =9.245,8.256,all P<0.05).Conclusion The effect of low molecular weight heparin in the treatment of hypercoagu-lable state of ulcerative colitis is effective , which can effectively reduce the inflammation level and hypercoagulable state of the patients ,it is beneficial and feasible for the prognosis of the patients .

While human induced pluripotent stem cells (hiPSCs) have promising applications in regenerative medicine, most of the hiPSC lines available today are not suitable for clinical applications due to contamination with nonhuman materials, such as sialic acid, and potential pathogens from animal-product-containing cell culture systems. Although several xeno-free cell culture systems have been established recently, their use of human fibroblasts as feeders reduces the clinical potential of hiPSCs due to batch-to-batch variation in the feeders and time-consuming preparation processes. In this study, we have developed a xeno-free and feeder-cell-free human embryonic stem cell (hESC)/hiPSC culture system using human plasma and human placenta extracts. The system maintains the self-renewing capacity and pluripotency of hESCs for more than 40 passages. Human iPSCs were also derived from human dermal fibroblasts using this culture system by overexpressing three transcription factors-Oct4, Sox2 and Nanog. The culture system developed here is inexpensive and suitable for large scale production.
Cell Culture Techniques ; methods ; Cell Differentiation ; Cellular Reprogramming ; Culture Media ; Extracellular Matrix Proteins ; isolation & purification ; Female ; Fibroblasts ; cytology ; Humans ; Lentivirus ; genetics ; Placenta ; chemistry ; Pluripotent Stem Cells ; cytology ; metabolism ; Pregnancy ; Sodium Chloride ; chemistry ; Transcription Factors ; genetics

Objective This study aimed to assess the efficiency and purification of the Trace DNA extraction with a quantified method for the magnetic bead-based DNA extraction as performed on the Tecan Automated systems with TE-MAGS magnetic separator. Methods Serial dilutions of standard commercial DNA 9947A were used with the total DNA contents, 0.1ng, 0.2ng,0.3ng, 0.4ng,0.5ng, 0.6ng, 0.7ng, 0.8ng, 0.9ng,1ng,diluted progressively and a 1ng DNA (standard commercial DNA 9947A) admixed with 6 common DNA-PCR inhibitors were extracted on the Automated systems and then performed via Fluorogenic probe quantitative PCR and STR genotype for the quantification analysis of recovery and purification. Results The recovery rate of standard 9947A DNA diluted with 0.1~1ng was 38.92~60.01%, and 0.3ng and more DNA could observed the full STR profiles. For the different PCR inhibitors, above 94.5% of bile acid, collagen and urea were efficient removal during the purification process, and the hemoglobin, melanin and humic acid removal efficiency were about 97.5%, 97.85%, 82.14%, respectively.Conclusion Our results suggested that The TE-MAGS magnetic bead-based DNA extraction was suitable for the extraction of trace DNA with high recovery efficiency and purification ability.

The aim of this study was to evaluate the roles of interleukin (IL)-17A in risk stratification and prognosis of patients with sepsis-associated acute kidney injury (SAKI). Methods: We enrolled 146 sepsis patients (84 non-SAKI and 62 SAKI patients) admitted to the emergency department from November 2020 to November 2021. Patients with SAKI were differentiated based on the severity of acute kidney injury. All clinical parameters were evaluated upon admission before administering antibiotic treatment. Inflammatory cytokines were assessed using flow cytometry and the Pylon 3D automated immunoassay system (ET Healthcare). In addition, a receiver operating characteristic (ROC) curve was utilized to determine the prognostic values of IL-17A in SAKI. Results: The levels of creatinine, IL-2, IL-4, IL-6, IL-17A, tumor necrosis factor alpha, C-reactive protein, and procalcitonin (PCT) were significantly higher in the SAKI group than in the non-SAKI group (p < 0.05). The level of IL-17A revealed significant differences among stages 1, 2, and 3 in SAKI patients (p < 0.05). The mean levels of PCT, IL-4, and IL-17A were significantly higher in the non-survival group than in the survival group in SAKI patients (p < 0.05). In addition, the area under the ROC curve of IL-17A was 0.811. Moreover, the IL-17A cutoff for differentiating survivors from non-survivors was 4.7 pg/mL, of which the sensitivity and specificity were 77.4% and 71.0%, respectively. Conclusion: Elevated levels of IL-17A could predict that SAKI patients are significantly prone to worsening kidney injury with higher mortality. The usefulness of IL-17A in treating SAKI requires further research.

Objective:To explore the effects of magnanimous therapy on the magnanimous and enterprising traits of lung cancer patients and the analysis of related factors.Methods:Totally 197 patients with lung cancer were divided into individual group ( n=62), team group ( n=75) and control group ( n=60). Comparison and correlation analysis were applied to the data before and after the electroencephalogram and the magnanimous questionnaire, the cancer response questionnaire, the T-type psychological scale, the cancer heart state questionnaire and the cancer patient's life function index scale. t test, analysis of variance and Pearson correlation analysis were processed by SPSS 23.0. Results:After treatment, the " enterprising" dimension and " magnanimous" dimension of individual group and the " enterprising" dimension of the team group ((3.035±0.309), (3.041±0.265), (3.173±0.371)) were higher than that before treatment((2.934±0.326), (2.908±0.315), (3.130±0.387), all P<0.05). There was negative correlation between " magnanimous" dimension of the magnanimous questionnaire and " subconscious" dimension of the T-type psychological scale in individual group( r=-0.280, P<0.05). In team group, the " enterprising" dimension of the magnanimous questionnaire was negatively correlated with " Psychological" and " Yield" dimension of the cancer heart state questionnaire( r=-0.279, -0.285, P<0.05), and positively correlated with " Facing" of the cancer response questionnaire, " Good physical condition and ability" and " Psychological well-being" dimension of the cancer patient's life function index scale( r=0.367, 0.402, 0.379, P<0.05). There was a negative correlation between the " enterprising" dimension of the magnanimous questionnaire and the beta wave value in individual group. Conclusion:The magnanimous therapy can improve enterprising and magnanimous level of patients with lung cancer, and the effects are related with the above-mentioned psychosomatic factors.

OBJECTIVETo explore the presence, frequency and clinical value of myeloid-derived suppressor cells (MDSCs) in peripheral blood of patients with small cell lung cancer (SCLC).

METHODSFlow cytometry using antibodies against CD11b, CD33, CD14 or HLA-DR was conducted to explore the unique cell surface markers of MDSCs and statistical analysis was performed to explore the correlation of MDSCs and clinical features.

RESULTSMDSCs were present in 36 patients with SCLC and uniquely marked by CD11b and CD33-positive, but HLA-DR-negative on cell surfaces and possessed mononuclear phenotype. The levels of CD11b(+)CD33(+)HLA-DR(-)cells (MDSCs) in the SCLC patients and healthy controls were (26.87 ± 6.87)% and (11.04 ± 3.76)%, respectively, with a statistically significant difference (P < 0.001). MDSCs level was significantly associated with clinical stage and tumor distant metastasis (P < 0.05) , but not with age, sex, smoking status and performance status. The later was the clinical stage, the higher was the MDSCs level (r = 0.665, P < 0.001). The level of MDSCs was higher in SCLC patients with distant metastasis than in those without metastasis (r = 0.489, P = 0.003). The level of MDSCs was higher before treatment than after treatment and the difference was statistically significant (P = 0.003).

CONCLUSIONSThe results of our study demonstrate the existence of MDSCs in SCLC patients and the MDSCs level is associated with SCLC stage, metastasis and treatments. MDSCs might be a novel biomarker for early diagnosis and prognosis for SCLC patients.

Flow Cytometry ; HLA-DR Antigens ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Myeloid Cells ; Phenotype ; Prognosis ; Small Cell Lung Carcinoma ; metabolism ; pathology

OBJECTIVE：To study the effects of Mahuang xixin fuzi decoction on Toll-like receptors （TLRs）response and cytochrome C oxidase （Cyt-CO）-mediated apoptosis regulation in mice with influenza disease of kidney-yang deficiency. METHODS：Totally 48 male Balb/c mice were randomly divided into normal group （n＝12）and modeling group （n＝36）. The modeling group was intraperitoneally injected with estradiol benzoate solution （8 mg/kg）and intranasally injected with influenza virus H 1N1（20 μL/mice）to establish the influenza disease compound model of kidney-yang deficiency. After modeling ，the mice were randomly divided into model group ，positive drug group （Oseltamivir phosphate capsules ，0.195 g/kg），Mahuang xixin fuzi decoction group （1.802 g/kg，by crude dru g），with 12 mice in each group. Each group was given relevant medicine intragastrically，normal group and model group were given， corresponding volume of normal saline intragastrically 20 mL/kg，once a day ，for consecutive 6 days. During admi-nistration，body weight and anal temperature of mice were mail：xsy407861520@163.com measured daily ；the percentage of initial body weight was calculated. After last medication ，the organ （spleen，thymus and lung ）indexes were calculated ；the pathological changes of lung tissue were observed. The viral load of influenza A virus H 1N1 in lung tissue was detected （reflected by M gene mRNA expression）；mRNA expressions of TLR3，TLR7，myeloid differentiation factor （MyD88）and Caspase- 3 in cardiac tissue as well as the activity of Cyt-CO and the content of cytochrome C （Cyt-C）were also determined. RESULTS ：Compared with normal group，initial body weight percentage and anal temperature of the model group continued to decrease （P＜0.05）；the spleen and thymus indexes were decreased significantly （P＜0.05），while lung index was increased significantly （P＜0.05）；the lung tissue lesions were serious. Viral load in lung tissue ，mRNA expressions of TLR 3，TLR7，MyD88 and Caspase- 3 in cardiac tissue as well as the content of Cyt-C were increased significantly （P＜0.05 or P＜0.01），while the activity of Cyt-CO in cardiac tissue was significantly decreased （P＜0.01）. Compared with model group ，initial body weight percentage and anal temperature of mice in Mahuang xixin fuzi decoction group showed an increasing trend from the fourth day of administration （P＜0.05 or P＜0.01）. The spleen and thymus indexes were increased significantly （P＜0.05），while the lung index was significantly decreased （P＜0.05）；the pathological injury of lung tissue was significantly improved ；viral load in lung tissue ，mRNA expressions of TLR 3 and Caspase- 3 as well as the content of Cyt-C in cardiac tissue were decreased significantly （P＜0.05 or P＜0.01），while the activity of Cyt-CO was increased significantly in cardiac tissue （P＜0.01）. CONCLUSIONS ：Mahuang xixin fuzi decoction can improve influenza disease of kidney-yang deficiency in mice ，the effect may related to inhibit TLRs response and apoptosis regulation pathway mediated by Cyt-CO.