Objective To investigate the relationship between the single nucleotide polymorphisms (SNP) of rs1008438 in HSP70 promoter and the susceptibility of high altitude pulmonary edema (HAPE) .Methods The PCR‐DNA sequencing method was used to analyze gene distribution of rs1008438 in 100 HAPE patients and 200 healthy people ,and the relationship between dif‐ferent genotypes and HAPE was evaluated .Meanwhile ,HSP70 protein in cytoplasm and nuclei of white blood cells were detected by ELISA in patientgroup and healthy group .TNF‐α,IL‐1β and IL‐6 levels were analyzed by protein chip technology and EVI‐DENCE180 automatic chip reader .Results The TT ,GT ,and GG genetype frequencies of rs1008438 in HAPE patients and healthy controls were 76 .0% ,20 .0% ,4 .0% ,and 93 .0% ,6 .5% ,0 .5% ,respectively .The TT genetype frequency in HAPE patients was significantly lower than that in healthy control(P<0 .05) .The HAPE incidence rate in GT/GG genetype was 4 .195 times higher than that in TT genetype ,and the allele G can also significantly increase the prevalence of HAPE compared to T allele (OR=4 .178) .ELISA showed that HSP70 were higher in HAPE group of all genotypes than those in healthy control of the same geno‐type .And the nuclei/cytoplasm ratio of HSP70 in TT genotype was higher than that in GT/GG genotype .Protein chip showed that the levels of TNF α,IL‐1β,and IL‐6 in HAPE group were significantly higher than those in healthy control .Conclusion The poly‐morphism of rs1008438 is related to susceptibility of HAPE .Mutate genetype may change the promoter activity and increase the ex‐pression of HSP70 ,which induced HAPE .

BACKGROUND: Soft tissue filling is a problem in clinic. It has been proved that ultrasonic wave-treated mature adipocytes cannot be used for cell transplantation.It is hopeful to solve the problem with adipose tissue engineering. OBJECTIVE: To observe the effect of ultrasonic wave on the in vitro culture and proliferation of preadipocytes, and validate the possibility of preadipocyte as seed cell in adipose tissue engineering following ultrasound-assisted liposuction. DESIGN: Controlled observation experiment. SETTING: Department of Plastic Surgery, Rennin Hospital, Wuhan University. MATERIALS: This experiment was carried out in the Medical College of Wuhan University from July 2003 to September 2004. One local 3-month-old hybridized pig was provided by the Experimental Animal Center of Medical College of Wuhan University. After the pig was anesthetized with ketamine, an area of 10 cm×20 cm was labeled on both sides of back respectively, and the tissue in the labeled area was harvested. The tissue on the right side served as experimental group and that on the left side served as control group. METHODS: The tissue of the expedmental group was pretreated for 8 minutes by ultrasonic wave with the energy of 3W/cm2. Under aseptic condition, the skin layer was open, and 100 g subcutaneous adipose tissue was resected from each side and then placed in prepared container containing cold D-hanks solution for later use. The ultrasonic wave-treated porcine preadipocytes of adipose tissue of experimental group were isolated and cultured in vitro, and the number and lipid content of preadipocytes were measured every other 2 days. Results were presented as the mean val ue of the number of cells of three wells. In the control group, pretreatment of ultrasonic wave was omitted. The growth curves of two groups were drawn. Intracellular adipose content was measured by oil red O staining. Absorbance (A) was measured with spectrophotometer (HITACHI G2000), which was regulated at the wavelength of 510 nm. MAIN OUTCOME MEASURES: ① Morphological observation of culture of porcine preadipocytes. ②The growth curves of experimental and control groups. ③3 Change in the lipid content of experimental and control groups.RESULTS: ① Cells in the control group adhered to the wall within 6 to 24 hours, and those in the experimental group basically adhered to the wall after 1 to 2 days, and there were many non-adhesive cells in the exoerimental group. Accumulation of adipose granules in the cells was found in the control group after 4 days and that was found in the experimental group after 6 days. On day 12, the cells in the control group basically differentiated into the cells with single lipid drop or multiple lipid drops, and a small amount of natant mature adipocytes were found, while few cells with lipid drop were found in the experimental group. ②Under the condition of the same amount of cells, the cell doubling time of experimental group was about 72 hours and that of control group was 36 hours. After 9-day culture, the number of cells in the experimental group and control group was 13×104 cells/well and 18×104 cells/well, respectively. The rate of cell proliferation of experimental group was very slow. ③Lipid of the experimental group appeared on the 6th day after culture, and that of the control group appeared on the 4th day after culture. The peak value of A was 0.32 and 0.68 in e penmental group and control group respectively.CONCLUSION: Ultrasonic wave has obviously inhibitory effect on the proliferation and differentiation of preadipocytes.The preadipocytes treated by ultrasonic wave are not suitable as seed cells.

Objective To study DNA damage induced by chlorinated drinking water by-product MX.Methods Hu-man hepatocellular carcinoma cell line HepG2cells was selected as target cells to test the DNA damage induced by MX using single cell gel electrophoresis assay.In this study,the cells were treated with MX at final concentrations of10,30,100and300?mol/L.DMSO and hydrogen peroxide were administered as solvent control and positive control respectively.Results MX induced DNA single strand breaks at the concentrations of30,100and300?mol/L.The length of DNA migration induced by MX increased significantly in comparison with the solvent control(DMSO)in a dose-response manner.Conclusions Chlorinated by-product MX in drinking water could induce obvious DNA damage to human hepatocellular carcinoma cell line HepG2cells.

Objective To understand the situation of college students' awareness about innovative credit, analyze the influencing factors, in order to provide a scientific basis for constructing innovation credits and promoting the double strategy. Method The undergraduates of a medical university who were still at school in April 2015 were selected, and by using a stratified cluster sampling method, 450 students were chosen to be conducted by self-administered questionnaire survey to understand the cognitive status of in-novation of credit system in college students. Chi square test was used to compare the passing rates of students in different demographic characteristics, and the relationship between scores was analyzed by linear regression analysis. Result A total of 439 valid questionnaires were collected , and the total scores of innovative knowledge, attitudes and behaviors of 439 undergraduates were 15.95％ (70/439), 36.44％ (160/439) and 32.80％(144/439) respectively. In the part of knowledge innovation credits, girls' passing rate was higher than boys ( χ2=4.010,P=0.045. In the part of attitude, high grade group students had more positive attitude than the low grade group of students ( χ2=6.227,P=0.0013). In the part of behavior; higher grade students with innovative credits had higher pass rates than those in lower grades (chi, 2=7.781, P=0.005), and boys had a higher rate of passing than girls ( χ2=6.658,P=0.010). The total score of knowledge was positively linear with the total score of attitude and behavior. Conclusion College Students' awareness of innovation credit rate is low, but the innovative attitude and behavior is positive. The higher the awareness rate is conducive to the cultivation of attitude and behavior, so it is necessary in medical colleges to carry out innovation of credit and promote the cultivation of innovative consciousness.

Objective To detect the expression of survivin protein in intramedullary gliomas and evaluated the clinical significance of the survivin expression. Method Seventeen cases of intramedullary gliomas were removed by using microsurgical technique.It composed of 8 cases of ependymomas and 9 ones of astrocytomas.The patients were followed up by MRI scanning periodically.The survivin protein expression of the intramedullary gliomas were examined by immunohistochemical stain (SP). Results Total resection of the tumor was obtained in 7 cases of ependymomas and subtotal resection was undertaken in the other one case.For 9 cases of astrocytomas,total resection of the tumor achieved in 3 cases,subtoal resection in 5 cases and partial resection in one case.Survivin expression was detected in 2 samples of ependymomas and 6 ones of astrocytomas.2 astrocytoma cases were moderate positive staining,who suffering from intracranial and vertebral subarachnoid dissemination of the tumor. Conclusion The result of microsurgical treatment for intramedullary ependymomas is satisfactory.The survivin expression in astrocytoma samples is significantly higher than that in ependymoma.Moderate positive staining may correlated with subarachnoid dissemination of the tumor.

Objective To clone and characterize Profilin encoding genes in Amaranthus spinosus and to analyze the contribution of different amino acids in isoallergens to allergen antigenicity and tertiary structure. Methods The primers were designed according to the core sequences which were obtained by bioinformatic analysis of the known Profilin amino acid sequences, followed by gene cloning from the Ama- ranthus spinosus cDNA pool and subsequent confirmation by double-digestion, colony PCR and DNA sequen- cing. Antigenicity evaluation and tertiary structural modeling of the encoded protein were accomplished by online software MULTIPRED and SWISS-MODEL, respectively. Results Two panallergenic genes, named as PRF7 and PRF23, were acquired from Amaranthus spinosus. Sequence and structure analysis demonstra- ted that there was some discrepancy in tertiary structures of the encoded proteins, besides distinct difference in their amino acid sequences. PRF7 exhibited high homology with panallergen Profilins Q64LH0, with the identities 98%, whereas the homology of PRF23 and Q9XF42 (apple allergen) was 81%. Q64LH0 and PRF23 were modeled as 3nulA (Q42449) and lg5uB (Q9LE18), respectively. PRF23 exhibited distinct0 three dimensional structural difference in certain fragments compared with Q64LH0 and other Profilins. Though the average values of antigenicity displayed no difference between Q64LH0 and PRF23 on whole se- quences, the antigenicity of PRF23 on certain fragments was obviously lower than that of Q64LHO because of the alteration of some amino acids with different characters, implying the cause of lower incidence of hay fe- ver in South China than in North China. Conclusion Based on sequence analysis, antigenicity evaluation and tertiary structural modeling for Q64LH0 and PRF23, we obtained lots of useful information about the contribution of different amino acids to antigenicity and protein structures, thus would facilitate allergen ge- netic improvement by amino acid replacement.

Objective To investigate the application of the parietal branches of superficial temporal artery island flap in the complex scalp defects.Methods A parietal branches of superficial temporal artery island flap on the ectatic scalp flap was designed to repair the complex scalp defects in 25cases and the repairing effect was observed.Results The island flaps were survived completely in 24patients,in which 1 patient had partial necrosis because of the flap tension was too large,but healed after local dressing and debridement.After followed up 6～ 12 months,the color and texture of the flap were the same to the surrounding normal scalp,and the shape was satisfactory.The flap donor site of hair growth was good,with well healing and no obvious complications.Conclusions The parietal branches of superficial temporal artery island flap can repair the complex scalp defects with the flexible flap design and movement.The flap survives well and the repair area is large.The flap and the surrounding scalp connects good.Therefore,it is a good method strongly recommended for small area complex scalp defects repair in clinics.

Objective To investigate the expression of sialic acid-binding immunoglobulin-like lectin-1 (Siglec-1) in the peripheral blood mononuclear cells (PBMCs) in patients with rheumatoid arthritis (RA),osteoarthritis (OA) and healthy controls and to explore the relationship between Siglec-1 expression and disease activity in RA.Methods Siglec-1 protein and mRNA levels were measured by flow cytometry and real-time quantitative reversetranscription-polymerase chain reaction (qRT-PCR) in 42 RA patients,28 OA patients and 26 healthy controls,respectively.The correlation studies between Siglec-1 and disease activity score 28 (DAS28) or C-reactive protein were performed.T-test was used for comparisons between groups and Pearson's correlation test was used for correlation analysis.Results The percentage of Siglec-1 positive cells of PBMCs in RA group [(15.2±7.6)％] was significantly higher than that in the OA group [(2.3 ±2.6)％] or healthy controls [(2.1±1.6)％,t=8.615,8.661; all P＜0.01].And the major cell type in PBMCs that expressed Siglec-1 was monocytes.The relative Siglec-1 mRNA expression in PBMCs in the RA group (3.4±1.5) was also significantly higher than that in the OA group (1.2±0.4) or healthy controls [(1.0± 0.4),t=3.446,3.966; all P＜0.05].But no significant differences of Siglec-1 protein and mRNA between the OA group and healthy controls were found.Furthermore,positive correlations between Siglec-1 protein and DAS28 or hs-CRP were found in RA patients (r=0.89,P＜0.01; r=0.48,P＜0.01).Conclusion PBMCs are activated which are characterized by elevated expression of Siglec-1 in RA patients.Circulating Siglec-1 may be considered as a potential noninvasive biomarker for monitoring disease activity in RA.

AIM: To investigate the possible mechanism of PRL-2 in invasive metastasis of tumors.METHODS: The PRL-2 vector was transfected into CL1 cell with lipofectamine reagent,the transfectants were selected by growth in the medium supplemented with G418.Zymographic analysis of metalloproteinases(MMPs) activity was performed,RT-PCR was used to determine the mRNA levels of(MMP-2),MMP-9,TIMP-1 and TIMP-2,the protein levels of(MMP-2),MMP-9,TIMP-1 and TIMP-2 were analyzed by Western blotting.The effects of the special inhibitor of PRL-2 on transfected cells were also observed.RESULTS: The stable cell line selected by G418 was identified by RT-PCR and Western blotting.More abundance of MMP-2,MMP-9 and its activated type were secreted by the CL-1-PRL-2 cells than untransfected cells and transfected vector cells(P

OBJECTIVETo investigate the effect and mechanism of Huanglian Jiedu Tang active fraction (HIJDTAF) on calcium overloading in neurons.

METHODCerebral ischemia was imitated by hypoxia/hypoglycemia damage on fetal rat neurons. Double wavelength fluorospectrophotometry was used to assay the content of calcium in neurons in order to evaluate the effect of HLJDTAF on calcium overloading. Neurons were treated with glutamic acid, potassium chloride (KCl), A23187, caffeine(CAF) and methacholine (Mch) to analysis the related mechanism of HLJDTAF on calcium overloading in neurons.

RESULTHLJDTAF 0.3, 0.15 g x k(-1) could remarkably inhibit the calcium overloading in neurons caused by hypoxia/hypoglycemia, glutamic acid, KCl and A23187. HLJDTAF 0.3 g x kg(-1) could inhibit the increasing of calcium caused by CAF and Mch in the presence of and in the absence of extra-calcium.

CONCLUSIONHLJDTAF could remarkably inhibit the calcium overloading in neurons after cerebral ischemia injury, it probably plays the function via several pathways.

Animals ; Caffeine ; pharmacology ; Calcimycin ; pharmacology ; Calcium ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glutamic Acid ; pharmacology ; Male ; Methacholine Chloride ; pharmacology ; Neurons ; cytology ; drug effects ; metabolism ; Potassium Chloride ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Fluorescence