Abnormalities, Multiple ; blood ; diagnosis ; metabolism ; physiopathology ; Child ; Diagnosis, Differential ; Dwarfism ; blood ; diagnosis ; metabolism ; physiopathology ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Silver-Russell Syndrome ; blood ; diagnosis ; metabolism ; physiopathology

OBJECTIVETo explore the relationship of CD44 expression or glycosylation and hepatocellular carcinoma (HCC) metastasis.

METHODSIHC, Quantum dots detection, RT-PCR, Western blot, Cellular immune fluorescence and MS-PCR were used to identify CD44 expression in HCC samples and a series of human HCC cell lines with different metastatic potentials. Lectin array was used to reveal the relationship of CD44v6 glycosylation and human HCC metastasis.

RESULTSImmunohistochemistry analysis showed that CD44v6 was mainly distributed on the cell membrane, while CD44S immunoreactivity was prominently in the cytoplasm, CD44v3 and CD44v4/5 were in cytoplasm on membrane. Among CD44S and those CD44 variants, only the expression of CD44v6 was higher in metastasis HCC samples as compared to that in non-metastasis group (x²=8.828, P less than 0.05). This result was also re-confirmed by the result of Quantum dots (t = 2.392, P < 0.05) and serum detection (t = 2.56, P < 0.05). We found completely methylation of CD44v6 gene in Hep3B and incomplete methylation in MHCC97H and MHCC97L cell lines with metastatic potentials. The lectin affinity assay indicated that lectin MAL, SNA and WGA showed more affinity to MHCC97H and MHCC97Lcell lines than that of the non-metastatic Hep3B cell lines.

CONCLUSIONSCD44v6 over-expression presents a positive correlation with HCC metastatic potential, which may be associated with DNA methylation level in promoter sequence. The increasing sialic acid modified glycan of CD44v6 might be related to HCC metastatic ability.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Methylation ; Glycosylation ; Humans ; Hyaluronan Receptors ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; pathology ; Promoter Regions, Genetic

Objective To observe the clinical efficacy of deproteinized calf blood extractive injection combined with hyperbaric oxygen in patients with severe traumatic brain injury.Methods 74 patients with severe traumatic brain injury from January 2013 to March 2015 in Wenzhou hospital of Chinese traditional medicine were randomly divided into observation group and control group, 37 cases in each group.The control group received hyperbaric oxygen on the basis of conventional therapy, the observation group received deproteinized calf blood extractive injection on the basis of control group.The Glasgow coma scale ( GCS) , Barthel, hs-CRP, TNF-α, IL-6 and prognosis were compared between two groups.Results The GCS score and Barthel index scores post-treatment in observation group were (13.67 ±1.73),(65.73 ±4.02) points, which were higher than (9.66 ±1.24), (50.69 ± 3.76) points in control group, and the difference was significant (P<0.05).The serum hs-CRP, TNF-α, IL-6 post-treatment in observation group were (4.55 ±0.76)mg/L,(1.21 ±0.05)μg/L,(0.21 ±0.01)μg/L, which were better than those of control group (6.43 ±1.01)mg/L,(1.36 ±0.06)μg/L,(0.28 ±0.02)μg/L (P<0.05).The rate of favorable prognosis in observation group was 48.65%, which was higher than that of control group (P<0.05).Conclusion The deproteinized calf blood extractive injection combined with hyperbaric oxygen has the exact efficacy, which was better than hyperbaric oxygen alone in the treatment of severe traumatic brain injury.

AIM: To clone EBV-LMP2A gene, construct and identify the recombinant retroviral vector and stable cell strains expressing EBV LMP2A. METHODS: The full-length EBV LMP2A gene was generated by RT-PCR amplification from B95.8 cells which contain complement nucleotide sequence of EBV LMP2A gene. The gene was ligated to T-vector and sequenced to construct retroviral vector consisting with LMP2A. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pGEZ-LMP2A and two auxiliary viral vectors pHIT456 and pHIT60 by lipofectAMINE2000. Viral titration was performed according to the instructions of the manufacturer. To establish L929 cell line stable expressing LMP2A, L929 cells were infected with recombinant retrovirus three times and selected by Zeocine. The Zeocine-resistant clones (L929/LMP2A) were screened for LMP2A expression by RT-PCR and Western blot. RESULTS: The recombinant retrovirus vector carrying LMP2A gene was constructed successfully. Transfection yield a titer of 5×10~8 infectious particles/L. The infected L929 cells were selected by Zeocine. Results of RT-PCR and Western blot indicated that L929 transgenetic cells could stably express EBV-LMP2A. CONCLUSION: The L929 cell line stably expressing LMP2A provides suitability for extraction of the LMP2A protein and preparations of the vaccine for the therapy of EBV-associated diseases.

Flexible matching has recently been proposed as a method of improving interactions efficiency. In this study, the concept of flexible matching has been introduced, and the applicability of this strategy has also been described based on the power calculation of interaction between HER-2 polymorphism and smoking with breast cancer. A large-sample approximation method is used to estimate the power and efficiency of gene-environment interactions. In the basic scenario, power of interaction between HER-2 polymorphism and smoking of unmatched case-control study appears to be 30% while in the frequency matching case-control study it is 56%. However, when increasing the smoking prevalence in controls, greater power can be obtained (power=74%). Conclusions: Flexible matching strategies can increase the power and efficiency of case-control studies to detect and estimate the gene-environment interactions when compared with traditional frequency matching and it is especially useful under those scenarios when low environmental exposure of population, adverse gene-environment interactions or less paired controls are seen. Optimal matching design should be made available by weighing the benefits and loss due to flexible matching.

Objective To observe the dynamic changes of neuron-specific enolase(NSE)mRNA and protein in offspring rats brain tissue with bilirubin encephalopathy and explore the pathological mechanism and its diagnostic value on bilirubin encephalopathy.Methods Seven-day postnatal Wistar rats were used for study.One hundred and twenty rats were divided into 2 groups randomly(control group and experimental group),which were respectively subdivided into 6 groups(6,12,24,48,72,96 h).The rats in control group were intraperitoneally administered physiological saline 0.5 mL,the rats in experimental groups were intraperitoneally administered bilirubin(200 mg/kg).Reverse transcription-polymerase chain reaction was used to analyze the dynamic changes of NSE mRNA expression at 6,12,24,48,72 and 96 h in brain tissue of rats with bilirubin encephalopathy.Immunohistochemistry was used to evaluate NSE protein expression in hippo-campi,cerebral cortex,thalamic and pallidus at different times.Results The expression of NSE mRNA significantly decreased in brain tissue of rats with bilirubin encephalopathy from 6 h to 96 h compared with the control groups.The expression of NSE protein in hippocampi decreased in offspring rats with bilirubin encephalopathy from 6 h to 96 h,but there were no differences compared with the control groups.The expression of NSE protein in cerebral cortex was significantly decreased in rats with bilirubin encephalopathy from 6 h to 96 h,there were significant differences compared with the control group.The expression of NSE protein in thalamic significantly decreased in rats with bilirubin encephalopathy from 6 h to 96 h,but there were significant differences between experimental groups and the control groups at 24 h and 72 h.The expression of NSE protein in pallidus significantly decreased in offspring rats with bilirubin encephalopathy from 6 h to 96 h,and there were significant differences compared with control groups.Conclusions The changing trends of expression of NSE mRNA were identical to those of NSE protein.NSE may reflect the degree of injury of neurogliocyte.It can serve as reliable index to determine bilirubin encephalopathy.

Outcome mapping ( OM) is a participant-oriented new method for project planning ,monitoring and evaluation .It is based on continuous learning and focuses on changes in behavior ,relationship and activities of persons ,groups and organizations .The use of OM can help to clarify the roles and assignments of the government ,managers, health workers , and other relevant personnel , making full use of the limited social resources and improving the efficiency and quality of health services .We introduce the principle and operation steps of OM with case simulation in health resource integration , to show its application in decision-making .

Outcome mapping (OM) is a participant-oriented new method for project planning,monitoring and evaluation.It is based on continuous learning and focuses on changes in behavior,relationship and activities of persons,groups and organizations.The use of OM can help to clarify the roles and assignments of the government,managers,health workers,and other relevant personnel,making full use of the limited social resources and improving the efficiency and quality of health services.We introduce the principle and operation steps of OM with case simulation in health resource integration,to show its application in decision-making.
Decision Making ; Outcome and Process Assessment (Health Care) ; methods ; Planning Techniques

Adult ; Aldosterone ; blood ; Angiotensin II ; blood ; Diet, Sodium-Restricted ; Female ; Humans ; Kidney ; physiopathology ; Kidney Function Tests ; Liver Cirrhosis ; blood ; physiopathology ; Male ; Middle Aged ; Renin ; blood ; Sodium ; administration & dosage

OBJECTIVETo investigate the association between the apoptosis genes CASP3(rs12108497) and CASP9 (rs4646018) polymorphisms and the risk of developing stomach cancer.

METHODSIn this population-based case-controlstudy, 278 cases with stomach cancer and 278 age (± 5 years), gender, and residential area matched controls were recruited. The genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The unconditional Logistic regression analysis was utilized to calculate the odds ratios (OR) and 95% confidence intervals (CI).

RESULTSThe individuals with TC, CC genotypes of rs12108497 locus had significantly increased risk of stomach cancer in comparison to those carrying TT genotype (OR= 1.45, 95% CI: 1.01-2.07 for TC; OR= 2.17, 95%CI: 1.15-4.08 for CC). However, the rs4646018 locus of CASP9 gene polymorphism was not related to stomach cancer risk. Compared with the subjects carrying the both low-risk genotypes, those carrying 1 or 2 high-risk genotypes had a noteworthy increased risk of stomach cancer (OR= 1.60, 95% CI: 1.12-2.30). The combined high-risk genotypes appeared to be more evident in subjects of male (OR= 1.62, 95% CI: 1.05-2.49), ever-smokers (OR= 1.87, 95%CI: 1.12-3.12), ever-drinkers (OR= 1.92, 95%CI: 1.02-3.65) and no family history of cancer (OR= 1.78, 95%CI: 1.18-2.68).

CONCLUSIONThe current findings suggest that the polymorphism of CASP3 rs12108497 might be associated with the risk of stomach cancer. However, the CASP9 rs4646018 polymorphism may not be related to the stomach cancer risk.

Aged ; Case-Control Studies ; Caspase 3 ; genetics ; Caspase 9 ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk Factors ; Stomach Neoplasms ; enzymology ; genetics