Objective To compare the advantages and disadvantages of internal jugular vein and subclavian vein puncture approach by clinical and anatomical ways so as to explore the best way of establishing a central venous access to guide clinical work .Methods The 120 patients who were admitted into our hospital from January 2014 to January 2016 were randomly divided into the internal jugular vein group and the subclavian vein group with 60 cases in each group according to puncture approach , and the puncture were operated by the same group of physicians .The learning curve ,puncture success rate ,completion time ,complication rates and other indicators of the two groups were evalua-ted.Results The operating completion time of internal jugular vein puncture group was (22.00 ±5.58)minutes,the success rate was 52%, the learning curve was 12 cases;while the subclavian vein puncture group was (11.60 ±2.67)minutes,93% and 3 cases respectively,and the differences were statistically significant between the two groups (P<0.05).The total complication rate of the internal jugular vein punc-ture group was 58%while the subclavian vein group was 12%, and the difference was statistically significant (P<0.05).Conclusion Dif-ficulty,safety and complication rates of subclavian vein puncture approach were better than those of the internal jugular vein puncture ap -proach.Therefore, subclavian vein puncture approach should be the first choice when there’s a need to establish central venous access for rescuing severe patients such as shock .

Objective To investigate the application value of apolipoprotein E (ApoE) genotyping by DNA microarray technology and the relationship between ApoE allelic frequency and serum ApoE levels in both healthy individuals and patients with Alzheimer ′s disease (AD).Methods This research is case-control study.DNA microarray was used to detect the ApoE genotypes of AD patients (n =280) and age-matched non-demented elderly control subjects ( n =230) .The cases and controls were collected in Guangzhou Huiai Hospital during July 2014 to September 2015.The accuracy of genotype results was verified by DNA sequencing.Serum ApoE levels were measured by immunoturbidimetric assay .The ApoE genotype distribution and the relationship between ApoE allelic frequency and serum ApoE levels were analyzed.The “t” test was used to compare the ApoE levels of AD patients and controls , variance analysis was used to analyze ApoE levels in the persons with different genotype .Results DNA microarray technology genotyping results were completely consistent with the results of DNA sequencing .In AD group, the ApoE genotype distribution were 2.9%(8 /280) for ε2ε3, 1.8% (5/280) for ε2ε4, 46.8% (131/280)for ε3ε3,45.4%(127 /280) for ε3ε4 and 3.1%(9 /280) for ε4ε4.While in the control group, the ApoE genotype distribution were 0.9%(2 /230) for ε2ε2, 12.6% (29/230)for ε2ε3, 1.3%(3 /230) forε2ε4, 70.0% (161 /230) for ε3ε3 and 15.2% (35 /230) for ε3ε4.The average serum concentrations of ApoE were (33.29 ±10.87)mg/L in AD patients and (41.28 ±10.95)mg/L in the controls.Among all participants, the average serum levels of ApoE were (50.86 ±6.21) mg/L for ε2 carriers, (38.78 ± 12.07)mg/L for ε3 carriers and (30.47 ±7.68)mg/L for ε4 carriers.In AD group,ApoE level of ε2, ε3,ε4 carriers is (50.31 ±9.08)mg/L, (38.30 ±7.60) mg/L and (32.86 ±5.93)mg/L respectively.In the control group, the ApoE level of ε2, ε3, ε4 carriers is (51.00 ±5.53)mg/L, (41.01 ±10.09)mg/L and (32.86 ±5.93)mg/L respectively.The ApoE levels of persons with different ApoE alleles are ε2 >ε3 >ε4. The difference is significant (F =89.6, P <0.05).However, the ApoE levels in persons with the same ApoE genotype between healthy individuals and AD patients have no significant difference ( t =0.981, 2.878 and 1.732 respectively, P >0.05) .Conclusions DNA microarray technology possesses high efficiency and favorable accuracy.The ε2 allele is associated with a higher ApoE concentration , ε3 allele with a mediate concentration and ε4 allele with a lowest concentration.Serum concentrations of ApoE showed no significant difference between AD patients and the healthy groups who have the same genotype .The primary cause of the low serum ApoE levels in AD patients is that the ApoE ε4 allelic frequencies of them are higher than that of the healthy persons.

Objective To investigate the sensitivity of different methods in the diagnosis of portal hypertension associated with liver cirrhosis. Methods 99 patients with portal hypertension associated liver cirrhosis, and 49 healthy subjects as control were enrolled in this study. Their portal and splenic vein widths, and the conditions of varicose vein in esophagus and gastric fundus were determined, and portal pressures were measured by radionuclide method. The positive percentage of different methods for the diagnosis of portal hypertension were analysed. Results Most of the patients with portal hypertension had the extension of portal and splenic vein in the fifferent areas and in various degrees. There were no significant differences in the degrees of portal and splenic vein extension among the patients with mild, moderate or severe esophageal varicose vein. The width of portal and splenic vein was not related with portal pressure. Radionuclide method was the most sensitive in diagnosing portal hypertension. Conclusion Portal hypertension can be diagnosed according to the conditions of vessels and spleen. The sensitivity of different methods in diagnosing portal hypertension were different, and that of radionuclide method was the highest. The width of portal and splenic vein had no close relation with portal pressure and the degree of esophageal varicose vein.

Objective To study the changes of lactic dehydrogenase (LDH) and ? hydroxybutyric acid (HBD) in atrophic gastritis type A(AAG) patients. Methods Auto biochemistry instrument was used to detect serum LDH and HBD. Hemoglobin, vitamin B 12 and gastrin were also examined routinely. Results All AAG patients had elevated levels of LDH and HBD in serum and 60 percent of patients had a rise of indirect bilirubin. In the first day after supplement with folic acid and vitamin B 12 , LDH and HBD decreased simultaneously by 30% along with a decrease of indirect bilirubin. The changes of LDH and HBD were earlier than changes of reticulocytes. Conclusions Serum LDH and HBD rise significantly in AAG patients. This might be associated with in situ hemolysis of bone marrow. And it may be a good index for differential diagnosis between AAG and atrophic gastritis type B.

Objective To investigate the relationship of apolipoprotein E (ApoE) allelic frequency and serum lipid levels in patients with Alzheimer′s disease (AD). Methods DNA microarray was used to detect the ApoE genotypes of AD patients (n = 200) and age-matched non-demented elderly control subjects (n = 159). Serum lipid levels was measured by Immunoturbidimetric assay at the same time. We analyzed the ApoE genotype distribution and the relationship of apolipoprotein E ( ApoE ) allelic frequency and serum lipid levels . Results The ApoE ε4 allelic frequencies (25.5%) in AD group is higher than that of the control group (7.9%) (P < 0.05). The ε2 allele was associated with a higher ApoE concentration, whereas with a mediate concentration in ε3 and the lowest concentration ( P < 0 . 05 ) in ε4 . Serum concentrations of ApoE showed no significant difference between AD patients and the healthy population who were with the same genotype (P > 0.05). Conclusion The ApoE levels are negatively related to ApoE ε4 allele frequency and have no significant differences with the same genotype in AD and the control group,which suggests that lower serum ApoE levels in AD patients is caused by higher ApoE ε4 allelic frequency in AD than in healthy population.

Objective To analyze the clinical and X-ray manifestations of sclerosing adenosis of the breast,and to evaluate the val-ue of preoperative differential diagnosis between the sclerosing adenosis and the breast cancer combined with pathology.Methods X-ray manifestations and clinical features of 25 patients with the sclerosing adenosis of the breast confirm by surgical pathology were analyzed retrospectively.Then the X-ray manifestations and clinical features were compared with the pathological results.Results Total 25 patients suffered from the sclerosing adenosis of the breast.Among them,2 cases were bilateral sclerosing adenosis,23 ca-ses were unilateral sclerosing adenosis,6 cases with epithelial atypical hyperplasia,4 cases with intraductal papilloma,7 cases with fibroadenoma nodule formation,and the rest with the pure sclerosing adenosis.X-ray manifestations:7 cases presented the medium density nodular shadows with blurred and irregular edges,1 6 cases showed the star-shaped shadows of which 6 cases were the“black stars”,3 cases of“white stars”.One in 7 cases can only see a star shadow,another was unclear or the form was varied.1 3 patients showed the calcification among 4 cases presented the small clusters of distribution with needlepoint-like calcification,2 cases of bulky curved calcification,and the rest with scattered and regional distribution of dot,needlepoint-like calcification.Conclusion Combined with clinical manifestations,the breast X-ray plays an important role in the differential diagnosis of sclerosing adenosis and the breast cancer.

Objective To obtain a single toxin component from the jellyfish Cyanea capillata and provide a foundation for the further study on bioactivity and function of the serine proteases from C.capillata.Methods Primers designed with restriction enzyme were used to amplify the coding region of cDNAs (CcSP1, CcSP2 and CcSP3).PCR fragments were ligated with the pET-24a( +) vector to construct the recombinant plasmids (pET24a-CcSP1, pET24a-CcSP2 and pET24a-CcSP3).After screening and identification,the recombinant plasmids were transformed into the Rosetta (DE3).plysS for protein expression.After induction with IPTG, SDS-PAGE and Western-blot were used to detect the expression of the recombinant proteins.Results SDS-PAGE showed that the proteins of rCcSP1, rCcSP2 and rCcSP3 were expressed in a single band at about 34 kDa, 42 kDa and 42kDa, respectively.Western-blot detection with anti-His antibody further confirmed that these recombinant proteins were His-tagged CcSP1, CcSP2 and CcSP3 fusion protein were obtained.Conclusion Prokaryotic recombinant plasmids of C.capillata serine proteases are contructed and recombinant proteins are obtained, which establishes the foundation for future study on the function of serine proteases from jellyfish.

Due to the particularity of marine ecological environment, many marine lives have developed and accumulated a large amount of biological molecules with special chemical structures and physiological activities, representing an important resource for the development of marine drugs. Generally, the marine antibacterial proteins are mainly identified from the marine microorganisms, invertebrates and fish.In this review, the progresses on marine antibacterial peptides and antimicrobial activity proteins are briefly summarized.

Objective To investigate the effect of B7-H3 on human hepatocellular carcinoma cell line HepG2 mediating regulation on human peripheral blood CD8+T cell activation,cell cycle and secretion of IL-17.Methods The expression of the B7-H3 on HepG2 cells was detected by RT-PCR and FCM respectively.B7-H3 was silenced by PGPU6/GFP/neo-B7-H3shRNA plasmid via cathodolyte liposome transfection method.CD8+T cells were sorted from healthy human peripheral blood with immunomagetic beads.The effect of HepG2 cells on activation,cell cycle and cytokine secretion of CD8+T cells which was stimulated by PHA or PMA respectively were analyzed by FCM.Results B7-H3 was highly expressed on HepG2 cells,and PGPU6/GFP/neo-B7-H3shRNA plasmid could effectively block down its expression.Otherwise,HepG2 cells could inhibit the expression of CD69,the early activation phenotype of T cell,blockade B7-H3 on HepG2 cells could significantly attenuate the inhibitory effects.Likewise,blockade B7-H3 on HepG2 cells apparently reversed the inhibitory effects of HepG2 cells on CD8+T cell cycle through down-regulating the cell number in G0/G1 phase and up-regulating the cell number in S phase;Moreover,HepG2 cells caused a sharp increase of IL-17 which was secreted by CD8+T cells and the level of IL-17 was further up-regulated after blocking down B7-H3.Conclusion HepG2 cells highly expressed B7-H3 that could promote the inhibitory the effect of HepG2 on expression of CD69 and cell cycle of CD8+T cells.HepG2 cells were able to up-regulate the level of IL-17 secreted by CD8+T cells,in which B7-H3 played an inhibitory role.

Objective Based on real-time PCR technique and ring primers,to establish a simple,accurate,cost-effective and easily standardized quantitative assay for quantification of HER2 mRNA,and apply to provide medication guidance for clinical tumor personalized molecular targeted therapy.Methods Screening reference gene which was stable expression in breast cancer,and optimizing the PCR reaction system.Then a real-time PCR with Eva Green for quantification of the mRNA expression levels of HER2 gene was developed.The specificity,sensitivity and reproducibility of the method were evaluated 87 specimens including 55 liquid nitrogen-frozen breast cancer tissues and 32 normal tissues were detected by the real-time quantitative reverse transcription (FQ RT)-PCR and immunohistochemistry(IHC).Results The standard curve of the method indicated a good linear relationship between the Ct value and the template concentration with the correlation coefficient being 0.997.The linear range of the system was from 101 to 106 copies/μl and the lower detection limit was 101 copies/μl.It had a high sensitivity and good specificity.The inter-assay coefficients of variation of HER and RPL37A genes were (5.93 ± 0.57)％ and (5.11 ± 0.59)％,(2.49 ±0.81)％ and (2.98 ±0.97)％ respectively.The intra-assay coefficients of variation were (5.76 ±0.58)％and (7.71 ±0.61)％,(3.75 ±0.76)％ and (4.40 ±0.96)％ respectively.Using the optimized FQ RTPCR system,HER2 gene of 87 specimens was quantificated.The sensitivity of the assay was 96.36％ (53/55),the specificity was 78.13％ (25/32),the positive predictive value was 88.33％ (53/60),the negative predictive value was 92.59％ (25/27),and the total coincidence rate between FQ RT-PCR and IHC was 89.66％ (78/87).The correlation of the results between the FQ RT-PCR and IHC was good (Kappa =0.770,P ＞ 0.05).Conclusions The method can quantify the mRNA expression levels of HER2 gene rapidly and cost-effictively with high sensitivity and specificity.It can be applied to clinical molecular diagnosis with attractive prospect.