Objective To discuss the therapeutic effect of kaluohuangna and anjiahuansuan in treating pa-tients with hemoptysis induced by pulmonary tuberculosis. Methods The therapeutic effect of kaluohuangna and an-jiahuansuan was compared. 163 eases of patients with hemoptysis of pulmonary tuberculosis were divided into group A ( n = 81 ) and B ( n = 82 ), and treated with kaluohuangna and anjiahuansuan respectively. Results Overall effec-tive rates of group A and B were 90. 1% ( 73/81 ) and 89.0% ( 73/82 ) respectively ( P > 0.05 ). The time in stop-ping blood in group A ( 5.8±2.9 ) h was shorter than in group B ( 6.0±3.0) h ( P > 0.05 ). Side effects of group A (0/163 ) were lower than group B(24/163 ) ( P < 0.05 ). Conclusions Kaluohuangrta is the first choice for treating patients with a little and middle hemoptysis induced by pulmonary tuberculosis, with less side effect.

Cell culture technology is the most commonly used method in the in vitro experiments at present. However, monolayer cell culture technology has been unable to meet the demand of the researchers. This is because that monolayer cell culture cannot mimic the cellular environment in which multiple cells interact with each other in the body. We cannot discuss the relationship of many cells, because we do not know the relationship between cells through a single kind of cell. So cell co-culture medicine arises at the historic moment for the demand. With the development of research method in recent years, cell co-culture method also has been improved in practice: from direct contact co-cultures to indirect contact co-cultures, from two-dimensional co-cultures to three-dimensional co-cultures. Cell co-culture method is closer to the human body. It is also more advantageous to study the interaction among cells. Nowadays, there are more researchers tend to select this method to study the physiological and pathological in vitro model, tissue engineering, and cell differentiation research. At the same time, it has become the focus of drug research and development, drug analysis, mechanism of drug action, and drug targets. This article will review the studies of cell co-culture method, summarize advantages and disadvantages of various methods, so as to promote improvement of cell culture methods, to build cells co-culture system that more close to human body, and build the in vitro model that simulate internal circulation of human body further.

AMOS-PCR and MLVA were carried out to identify the Brucella strains isolated in Fujian province, which were classified as B. melitensis biovar 2 or 3 by conventional microbiological tests. All these 3 isolates were identified to be B. melitensis by AMOS-PCR. The genetic patterns obtained by MLVA were queried in the Brucella 2007 database and clustered with B. melitensis strains. It is evident that these two molecular assays may be used as the assistant tools in the identification of Brucella strains.

BACKGROUND: Adipose tissue-derived mesenchymal stem cells (ADMSCs) have the multilineage differentiation potential, and are relatively easier to be obtained, thus they have attracted more and more attention as a new seed cell for cell engineering.OBJECTIVE: To observe the in vitro culture conditions of ADMSCs isolated from rat's subcutaneous adipose tissue, and identify them using immunohistochemical staining.DESIGN: An animal experiment.SETTING; Department of Cardiology, the General Hospital of Chinese PLA.MATERIALS: One healthy male Wistar rat of clean degree, 4 months old, weighing 200 g, was used. DMEM, fetal bovine serum were from GIBCO; Monoclone antibodies of rabbit-anti-rat CD13, CD34, CD44, CD45, CD105, D-related human leucocyte antigen (HLA-DR), factor-Ⅷ, vov Willebrand factor (VWF), Myosin, SABC kits and DAB staining kit from Wuhan Booster Biological Engineering, Co.,Ltd; Adeno-associated virus encoding green fluorescent protein from Vector Gene Technology Company Limited (Beijing).METHODS: The experiments were carried out in the Department of Internal Medicine, the General Hospital of Chinese PLA in October 2006. ① Cell isolation and culture: 0.3 g adipose tissue was cut from subcutaneous adipose tissue of Wistar rat's groin under aseptic condition, then minced and digested before culture, DMEM was changed at 2-3 days after plenty of fusiform-shap ed attached cells were observed under microscope, and the cell growth was observed. The cell concentration was adjusted to 2×107 L-1, then seeded into 96-well plate, and 100 μL for each well. From the second day, 3 wells were randomly selected every day, the cells were released with tripsin, and counted with blood cell counting chamber under inverted microscope. ② Cell viability assay: ADMSCs of passages 3 to 8 were added to DMSO freeze medium, and thawed after 2-4 weeks, and the cell viability was assessed by trypan blue dye exclusion. ③Immunohistochemical staining and identification: 2 ×107 L -1 cells were seeded to culture plate, then the immunohistochemical (SABC method) identification and Oil red O staining were performed to determine the cell surface antibodies of CD13, CD34, CD44, CD45, CD105, HLA-DR, factor-Ⅷ, HLA-DR and VWF. ④Lineage-specific differentiation and identification: The ADMSCs were plated on multi-well chamber and induced with lineage-specific media supplementation at least two weeks and identified by histologic/immunohistochemical assay of Oil red O for adipogenisis, alkaline phosphatase (ALP) stain for osteogenisis and Myosin monoclonal antibody for myogenisis. ⑤Transfected adenovirus carried green fluorescence protein (AD-GFP) medium: The fourth generation of ADMSCs were seeded on 96-well plate, 3 000 cells for each well, serum-free DMEM was changed after 24 hours, and added by AD-GFP at the same time, then transfected with different multiplicity of infection (MOI) of 1∶50, 1∶100, 1∶150 and 1∶200respectively, and then the transfection was observed.MAIN OUTCOME MEASURES: ① Results of cell isolation and culture; ② Cell viability after freezing and thawing; ③Results of immunohistochemical staining and identification; ④ Results of lineage-specific differentiation and identification;⑤ Results of transfected adenovirus carried AD-GFP.RESULTS: ① About 3.6×105 attached cells were obtained from 0.3 g subcutaneous adipose tissue, and these cells could be subcultured for passages in vitro with stable population doubling time. ② The cells were thawed after freezing for 2-3 weeks, and the trypan blue staining showed that the cell viability was above 90%. ③ The immunocytochemical staining showed that CD13, CD44, CD105 were positive and CD45, factor-Ⅷ, HLA-DR and VWF negative in different generations. ④ From the second generation, a few Oil red O positively stained cells were observed, which were obviously increased after prolonging the refreshing. After lineage-specific differentiation, the cells were all positive by Oil red O staining, ALP staining and Myosin immunohistochemical staining. ⑤ 72 hours after transfection, it was observed under fluorescence microscope that most cells were green fluorescence when the MOI value was 1∶200, the transfection was successful, and it was generally determined that the transfection rate was above 90%.CONCLUSION: A large number of ADMSCs with multilineage differentiation potential can be easily obtained from rat adipose tissue, osteoblast, myoblasts, they can be expanded in large quantity and stored in vitro for long time, AD-GFP were also successfully transfected.

Objective To investigate the relationship between cardiovascular risk factors and arterial elasticity in elderly hypertensive patients.Methods 253 subjects were divided into two groups:hypertension group (n=141) and non-hypertension group (n =112).Carotid-femoral pulse wave velocity (C-FPWV),carotid-radial PWV (C-RPWV),height and weight were determined.Fasting plasma levels of blood glucose (BG),blood lipids,insulin,endothelins (ET),nitric oxide (NO) and high sensitivity C-reactive protein (hsCRP) were assessed.Results C-FPWV and CRPWV were higher in hypertension group than in non-hypertension group [(11.7±1.9) m/s vs.(9.7±1.1)m/s,(11.7±1.7) m/s vs.(9.4±1.1)m/s,t=8.43 and-6.30,both P＜0.01].Body mass index (BMI),systolic blood pressure (SBP),diastolic blood pressure (DBP),levels of triglyceride (TG),BG,ET,NO and hsCRP,HOMA-IR had significant differences between the two groups (t=-5.27,-4.18,-6.00,6.29,-4.18,-4.86,-3.41,respectively,all P＜0.05).Pearson correlation analysis showed that CFPWV was correlated with SBP,age,DBP,fasting BG,NO,ET,TG,HOMA IR,cholesterol (CHO),hsCRP (r=0.534,0.374,0.340,0.338,-0.306,0.242,0.228,0.225,-0.218,0.178,respectively,all P＜0.05); C-RPWV was correlated with age,DBP,SBP,NO,TG (r=0.312,0.319,0.241,-0.197,0.151,respectively,all P＜0.05).Multiple regression analysis demonstrated that C-FPWV=4.640 + 0.081× HOMA IR+ 0.047× age+0.035×SBP+0.29×TG-0.017×NO+0.014×ET+0.132×hsCRP,r2=0.514; CRPWV=3.161+0.034×age+0.033×DBP+0.313×fastingBG+0.013×ET-0.013×NO,r2=0.390.Conclusions The decreases of arterial elasticity are closely related with endothelial function,insulin resistance,levels of blood lipids and hsCRP in elderly patients with hypertension.

Objective To investigate the correlation between dose and effect of thioacetamide (TAA) on rat model of intestinal endotoxemia. Methods The models of intestinal endotoxemia were induced by three different doses of TAA by gavage administration of TAA 200, 400, 600mg/kg respectively once per day for two days.The doses were given at same time point every day. Each group included 10 rats. The rats in the control group were administrated with 2 mL 0.9% NaCl saline gavage. The death of the rats was observed at 24 hours and 48 hours after administration. The blood samples of the living rats were drawn from abdominal aorta for determining the plasma endotoxin levels, serum alanine aminotransferase(ALT)and aspartated transaminase (AST) levels. The histopathological changes of liver were examined. Single factor analysis of variance was performed and comparision between groups was done using t test. Results No rat in the control group died. Two rats of 200 mg/kg TAA group, five rats of 400 mg/kg TAA group and eight rats of 600 mg/kg TAA group died during the experiment. The mean serum ALT levels of TAA model groups [(305.09±116.78)U/L,(901.67±274.31)U/L,(1454.84±473.49)U/L] were all significantly higher than that of the control group(47.81±22.61)U/L(t=14.583, 25.896 and 20.596, respectively; all P＜0.05). The mean serum AST levels of TAA model groups [(465.88±139.96)U/L, (884.37±250.90)U/L,(1889.23±159.67)U/L] were all significantly higher than that of the control group (69.33±22.04)U/L(t=12.988,18. 455 and 13.542, respectively; all P＜0.05). The mean plasma endotoxin levels of TAA model groups [(0.436±0.110)EU/mL, (0.550±0.095) EU/mL, (0.620±0.057)EU/mL] were all significantly higher than that of the control group (0.103±0.056)EU/mL(t=7.335, 5.260 and 8.191, respectively; all P＜0.05). The histological results of TAA model groups showed hepatic cell degeneration and necrosis in different degrees. Conclusions TAA with 200-600mg/kg is proper to establish the rat model of intestinal endotoxemia. The death rate of rats in the 200mg/kg TAA group is lower than those of other model groups, which suggests that 200mg/kg TAA may be the best dosage for establishing rat model for further studies.

Objective To test experimentally Fluoro-Jade B(FJB) stain method for detecting degeneration of neurons in the basal ganglia. Methods Kainic acid(KA)-lesion model by stereotaxical injection of KA into striatum of rats,MPTP-lesion model by injection of MPTP into intraperitoneal cavity of mice,as well as KA-lesion model of cultured striatal cells were firstly prepared.FJB stain dye was then used to visualize degeneration of neurons in above KA-or MPTP-lesion models. Results KA-or MPTP-induced degenerative neurons including cell bodies and processes could be clearly visualized by FJB stain dye.In the brain sections,FJB-positive stained degenerative neurons were numerously observed in the striatum of KA-lesion rats and the substantia nigra pars compacta of MPTP-treated mice,but not detected in the control animals.Moreover,degenerative neurons were also detected with FJB stain in cultured striatal neurons.Semi-quantitative analysis on percentage(?s) of FJB-positive neurons constituting total cultured striatal neurons in unit area showed that degenerative neurons of KA-lesion group (8.42?1.09)% was evidently more than that of controls (3.42?0.45)%,P

Objective To summarize the nursing cooperation during anterior cruciate ligament(ACL)reconstruction with autogenous quadurpled hamstring tendons under arthroscopic guide.Method Data of 25 cases of ACL reconstruction with autogenous quadrupled hamstring tendons under arthroscopic guide during January 2008 and December 2012 were analyzed and summarized the key points retrospectively.Result All the cases of ACL reconstruction were completed smoothly without any complication.Conclusion Excellent nursing cooperation including well-done preoperative preparation of instruments and accurate surgical cooperation are a guarantee of successful operation.

Objective:To examine the occurrence of seizures and perioperative risk factors in children after obtaining cardiopulmonary bypass (CPB).Methods:Electroencephalograph (EEG) and cerebral oxygen saturation (ScO 2) were monitored in 87 patients (aged 3 days-110 months, with the median at 4.4 months) and they were admitted to the Cardiac Intensive Care Unit (CICU) from May 2018 to April 2019, with 77 patients for 72 h after CPB and 10 cases with clinical seizures after CICU discharge for 8-180 h. Clinical data were recorded every 4 hours, including blood pressure, central venous pressure (CVP), temperature, arterial blood gases, lactate, doses of Dopamine, Milrinone, and Adrenalin, and daily C-reaction protein (CRP) and N-terminal pro-brain natriuretic peptide(NT-proBNP). Demographic data, the Society of Thoracic Surgeons-European Association for Cardio-Thoracic Surgery(STAT) mortality classification and clinical outcomes (duration of mechanical ventilation, CICU and hospital stay, and death) were recorded. Results:Seizures occurred in 24 patients (27.6%) within 48 h after CPB.EEG seizures appeared in 7 patients, both EEG and clinical seizures in 7 patients, and clinical seizures in 10 patients.Three patients developed epilepsy.There were 2 hospital deaths and both had EEG seizures.Patients with seizures had higher STAT categories, longer operative time and worse clinical outcomes (all P<0.05). Univariate regression revealed that the occurrence of seizures was positively related to adrenalin, temperature, lactate level (all P<0.01), blood pressure, CVP (all P<0.05) and CRP ( P<0.1), and negatively correlated with arterial carbon dioxide tension[ pa(CO 2)], ScO 2 and dose of Milrinone (all P<0.05). In multivariate regression, only dose of Adrenalin ( P<0.001) and lactate level ( P<0.01) remained significant. Conclusions:The incidence of seizures was high in children after CPB.Numerous perioperative factors that were related to seizures were identified.The most significant risk factors were higher dose of adrenaline and lactate level.Studies on neurological injury after CPB are urgently needed in China in order to reduce brain injury and improve neurodevelopmental outcomes.

Objective To analyze and discuss the effect of heat preservation on the recovery time of BIS and the concentration of propofol in the recovery period of the elderly patients with general anesthesia in order to provide guidance for clinical treatment.Methods78 elderly patients with general anesthesia were selected as the subjects, and the patients were divided into the control group and the study group with 39 cases in each group.The patients were divided into the control group and the study group.Control group, only to be liquid input and covered by drapes, research group take liquid input and peritoneal flushing fluid heating and heating blanket coverage of insulation measures, were recorded and compared between the two groups from anesthesia began to operation to complete the different time points of esophageal temperature and MAP, and to observe the recovery time of patients and effect compartment concentration differences.ResultsTwo groups at T0, T1 esophageal temperature no significant difference, study group t2-t6 esophageal temperatures were significantly higher than those of the control group (P<0.05), maps of the other two groups at different time points had no significant difference;when two groups of patients with propofol withdrawal BIS values had no significant difference (P<0.05), and in the time of discontinuation BIS is more than or equal to 80 recovery time and effect compartment concentration have significant difference (P<0.05).ConclusionThe surgical treatment of elderly patients with general anesthesia can help maintain the body temperature, shorten the recovery time of BIS and reduce the concentration of propofol in the recovery period.