Adult ; Critical Illness ; Extracorporeal Membrane Oxygenation ; methods ; Female ; Humans ; Myocarditis ; therapy

With the advantage of clear target and little side effect, antibody drug has attracted widely attention of worldwide pharmaceutical companies. However, large scale mammalian cell culture and antibody quality analysis are the bottlenecks of antibody drug industrialization in China. Especially due to the significant effect of cell culture conditions on antibody heterogeneity. Therefore, it is extremely urgent to optimize cell culture conditions to favor the demands of antibody drug development. This review summarized the most recent advances in the effect of cell culture conditions on antibody quality, followed by addressing the key issues that might be strategically important for domestic antibody drug development.
Animals ; Antibodies, Monoclonal ; biosynthesis ; isolation & purification ; Antibody Formation ; Cell Culture Techniques ; methods ; trends ; Genetic Heterogeneity ; Protein Stability ; Quality Control

BACKGROUND:Neural stem cells transplanted into the infarct region can promote the recovery of damaged nerve cells after cerebral infarction.
OBJECTIVE:To analyze the relevant influential factors of neural stem celltransplantation in the treatment of cerebral infarction.
METHODS:In this paper, we analyzed the experimental studies concerning neural stem celltransplantation for cerebral infarction published in recent years based on database search. There were main two aspects for discussing the progress in neural stem cells and cerebral infarction, the Chinese clinical trial registration and basic experimental studies.
RESULTS AND CONCLUSION:After cerebral infarction, neural stem cellproliferation and differentiation is closely related to the brain microenvironment. Large amount of nerve cellloss can be found in the infarct region. Cytokines can play a role in the neural stem celltransplantation to repair neurological injury after cerebral infarction, and also can induce neural stem cellproliferation, differentiation and migration, including epidermal growth factor, brain-derived neurotrophic factor, insulin-like growth factor 1, nerve growth factor, and basic fibroblast growth factor. Acupuncture and traditional Chinese medicine can promote the proliferation, migration and differentiation neural stem cells in the subependymal zone after cerebral infarction. Neural stem celltransplantation in the treatment of cerebral infarction has yielded progress, but there are stil many issues that need to be resolved in the future.

Objective To assess the mortality and risk factors for stroke among dialysis patients with different dialysis modality. Methods 590 patients who underwent hemodialysis (HD) or peritoneal dialysis (PD) from January 2008 to December 2012 were recruited in our study, and categorized according to dialysis modality. The prognostic risks of stroke were hazard ratio of risk was calculated by Cox regression analysis in HD and PD patients respectively. by the Kaplan?Meier curves or the Cox proportional hazards model. Results A total of 590 patients is under a median follow?up of 32.5 months. The stroke incidence rate of 49.2/1, 000 patient?years in total patients, and 74.1/1, 000 patient?years in HD patients, which was significantly higher compared with that of 31.8/1,000 patient?years in PD patients. On multivariate analysis, independent predictors of stroke occurrence were age(HR=1.05;95%CI:1.02~1.09;P=0.003)、diabete(HR=1.98;95%CI:1.31~3.46;P=0.001)、CVD(HR=2.06;95%CI:1.62-3.05;P < 0.001)、Total triglycerides(HR = 1.20; 95% CI:1.08-1.58; P = 0.034) and hemodialysis (HR = 2.03; 95% CI:1.46-3.89; P = 0.005). Conclusions Age, diabete, CVD, total triglycerides and hemodialysis are independently associated with increased stroke risks in dialysis patients, which suggest that these patients should pay attention to weight control and glucose control.

OBJECTIVE: To investigate biological characteristics of human umbilical cord-derived mesenchymal stem cells, and to explore the possibility of hepatocyte-like cells differentiation.METHODS: The umbilical cord was provided by healthy term birth woman in Tianjin Third Central Hospital. Mesenchymal stem cells were isolated from human umbilical cord by enzyme digestion method. Cells were passaged at 80%-90% confluent. The ninth passage of cells at a density of 5×10~(10)/L were seeded in 12-well culture plate and incubated with DMEM containing hepatocyte growth factor, fibroblast growth factor-4 and oncostatin for 28 days. Cell growth activity was detected by MTT method; cell cycle was detected by flow cytometry; surface immunological marker in MSC was detected by immunocytochemical stain and flow cytometry; specific surface phenotype of hepatocyte was detected by immunocytochemical staining. Function characteristic of hepatocyte was determined by staining for glycogen.RESULTS: MSCs were isolated from human umbilical cord and presented with fibroblastic morphology. 80% of cells were at G_0/G_1 phase with good growth activity and stably passaged over 20 times. These cells were positive for CD29, CD105, and Vimentin, but negative for CD34 and CD31. MSCs were induced to hepatocyte-1 ike cells that were positive for alpha fetoprotein, CK18, CK19 at 1 week and albumin at 3 weeks. At 4 weeks, induced cells were positive for glycogen staining.CONCLUSION: MSCs isolated from human umbilical cord can be cultured in a long periods time in vitro and are able to differentiate into functional hepatocyte-like cells.

Objective To investigate the effect of Shanzha Xiaozhi capslue on intima-media thickness(IMT) and plaques score of carotid artery in patients with non-acute phase coronary heart disease and traditional Chinese medicine(TCM)syndrome of phlegm and blood stasis. Methods A prospective study was conducted to carry out a research on 102 patients with non-acute phase coronary heart disease and TCM syndrome of phlegm and blood stasis. They were randomly divided into two groups:a control group(50 cases)treated with conventional western medicine alone and a observation group(52 cases)which was treated by both conventional western medicine and Shanzha Xiaozhi capslue(the main TCM ingredients:Shanzha,Dahuang)0.7 g,3 times a day,the therapeutic course being 6 months in both groups. The cardiocerebral vascular incidences of the two groups were observed,in the mean time,the carotid artery IMT and plagues score were registered,the scores of phlegm stagnation syndrome and blood-stasis syndrome were measured,adverse reaction was observed and compared in both groups before and after treatment. Results After treatment,the occurrence of major adverse acute cardiovascular and cerebrovascular events in observation group were reduced significantly compared with those in the control group(5.77%vs. 20.00%,P<0.05). The IMT and plaques score of carotid artery and the scores of phlegm stagnation syndrome and blood-stasis syndrome were all decreased obviously compared to those before treatment,and the effect was more remarkable in the treatment group〔IMT of carotid artery(mm):0.80±0.13 vs. 0.95±0.12,the plaques score:1.35±0.65 vs. 1.75±0.88, phlegm syndrome score:20.98±6.42 vs. 35.55±9.22,blood-stasis syndrome score:23.23±5.12 vs. 28.95±6.38, P<0.05 or P<0.01〕. Abdominal pain appeared in 2 patients of observation group without other adverse reactions. Conclusion Shanzha Xiaozhi capsule can stabilize atherosclerotic plaque and reduce the occurrence of acute adverse cardiocerebral vascular events in patients with non-acute phase coronary heart disease and phlegm and blood stasis syndrome possibly by removing phlegm and blood stasis to decrease the IMT and plaques score of carotid artery.

A double-molecular beacons (DMB) based assay was developed for porcine circovirus 2(PCV2) detection. Two single-stranded DNA molecular beacons which could specifically hybridize with PCV2 genome DNA respectively in different sequence were designed according to the characteristics of the PCV2 genome sequences. The fluorescence signal was amplified 80 times by DMB, which was 2-4 times higher than that of single molecular beacon. Under the optimal conditions of 10 mmol/L MgCl2 , 20 mmol/L Tris-HCl (pH=8. 0), 40 ℃ and 30 min incubation time of DNA with DMB, the enlargement factor was increased linearly with DNA concentration over the range from 2 nmol/L to 200 nmol/L, with a detection limit of 1 nmol/L. The method was applied to detect PCV2 in genome of 18 swine fever samples and 8 PCV2 positive cases were found, which were confirmed by PCR method.

OBJECTIVE A surveillance study was performed for nosocomial infections in order to investigate the change in antimicrobial resistance of Klebsiella pneumoniae,especially the strains isolated from 1999 to 2004.METHODS K-B test was used for the antibiotics susceptibility test and the results were read based on National Committee for Clinical Laboratory Standards(NCCLS) of the USA.The situation of ESBLs-producing strains of K.pneumoniae was investigated.RESULTS Totally 326 K.pneumoniae strains showed the highest susceptibility to imipenem.Ceftazidime,cefepime,and cefoperazone/sulbactam also showed excellent activity against K.pneumoniae.The prevalence of ESBLs from 326 strains was 20.2%.CONCLUSIONS It is important to study the drug resistance in nosocomial infections by K.pneumoniae.

Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.

Objective: To analyze the long-term prognosis of re-operation in patients with functional tricuspid regurgitation (FTR) after left sided valve replacement (LSVR) and hence evaluate the optimal timing of mentioned re-operation. Methods: A total of 59 FTR patients who had re-operation after their prior LSVR in our hospital from 1999-01 to 2013-01 were analyzed. The clinical information and post-operative follow-up results were recorded in all patients. Results: There were 5/59 (8.5%) patients died in peri-operative period and the overall post-operative mortality was 11.9% (7/59). The follow-up data of 54 survivors were available for the mean time of 51.1 (21-188) months. There were 19/54 (35.2%) patients suffered from MACE and 30 (55.6%) were beneifted by improved cardiac function. Uni-variable analysis indicated that pre-operative NYHA class IV (P=0.008), pre-operative right ventricular (RV) dysfunction (P=0.037), concomitant left-sided redo-operation (P=0.017) and TVR operation (P=0.002) were associated with all cause mortality of tricuspid re-operation. Multi-variable Cox regression analysis showed that pre-operative RV dysfunction was the only independent risk factor of long term MACE-free accumulating survival rate (HR=3.0, 95% CI 1.11-8.2,P=0.031); while TVR operation (HR=12.8, 95% CI 1.53-107.02,P=0.019) and pre-operative NYHA class IV (HR=5.3, 95% CI 1.20-24.51,P=0.032) were the independent risk factors for long-term mortality in patients after tricuspid re-operation. Conclusion: Patients with compensatory RV function showed better long term prognosis after secondary tricuspid operation. Aggressive re-operation before the occurrence of right ventricular dysfunction could be beneficial for relevant patients.