OBJECTIVETo explore the expression patterns of the chemokine CXCL12 and its receptor CXCR4 in human sperm.

METHODSWe collected semen samples from 10 fertile men, performed density gradient centrifugation, and then determined the expressions of both CXCL12 and CXCR4 in the sperm by RT-PCR and immunofluorescence staining.

RESULTSRT-PCR revealed the mRNA expressions of CXCL12 (0.641 +/- 0.180) and CXCR4 (0.464 +/- 0.100) in the sperm. However, only CXCR4 rather than CXCL12 was expressed at the protein level, and the positive staining for CXCR4 was observed mainly in the posterior part of the acrosome.

CONCLUSIONCXCL12 and CXCR4 are involved as important molecules in regulating the function of human sperm.

Acrosome ; metabolism ; Centrifugation, Density Gradient ; Chemokine CXCL12 ; metabolism ; Humans ; Male ; Receptors, CXCR4 ; metabolism ; Signal Transduction ; Spermatozoa ; metabolism

Objective To investigate the role of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) in inducing insulin resistance in differentiated human adipocytes.Methods Human preadipocytes obtained via liposuction were induced to differentiate into mature adipocytes.iE-DAP,a specific ligand for NOD1,was administered to human adipocytes in culture.NF-κB transcriptional activity and proinflammatory cytokines production were determined by luciferase assay and enzyme-linked immunosorbent assay.Glucose uptake in adipocytes was measured by 2-deoxy-D-[3 H] glucose uptake (P＜0.05).The expression of phosphatidylinositol-3-kinase p85 (PI-3K p85),insulin receptor substrate-1 (IRS-1) and Akt phosphorylations were detected by Western blotting.Results NF-κB transcriptional activity and cytokines such as interleukin (IL)-6,IL-8,and monocyte chemotactic protein 1 secretion were markedly increased after stimulation with iE-DAP (P＜0.05 or P＜0.01).Insulin-induced glucose uptake was decreased with the activation of NOD1 in a dose-and time-dependent fashion.NOD1 activation weakened insulin signal transduction as being revealed by increasing IRS-1 Ser307phosphorylation,reducing protein expression of PI-3K p85,and attenuating insulin-induced phosphorylation of Akt on Ser473 and Thr308in human adipocytes.Conclusion These results indicate that NOD1 activation induces inflammatory response and insulin resistance via IRS-1/PI3-K/Akt signaling pathway in human adipocytes.

OBJECTIVE To investigate the existence of genes for beta-lactam antibiotic resistance and for aminoglycosides modification enzymes(AMEs) in Pseudomonas aeruginosa(PAE) isolates from ICU patients and analyze the homology among strains.METHODS ?-Lactamase genes including TEM,SHV,OXA-10,PER,VEB,GES,CARB,IMP,VIM,SPM,GIM,DHA,FOX,MOX and oprD2,were detected by PCR amplication in 21 PAE isolates.The genes for AMES including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰwere determined by PCR amplification as well.RESULTS Among 21 isolates 21(100%),2(9.5%),1(4.8%),2(9.5%)and 4(19.0%) were positive for TEM,SHV,GES,CARB and VIM genes,respectively.The deletion of oprD2 gene was found in 14 out of 21 strains.Other ?-lactamase genes were absent in all isolates.As for AME genes,aac(3)-Ⅱ,aac(6″)-Ⅰ,aac(6)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aac(3)-Ⅰgenes were present in 19.0%,23.8%,9.5%,4.8%,and 19.0% of 21 isolates,However,aac(3)-Ⅰ gene was no position in any isolates.CONCLUSIONS P.aeruginosa carries various beta-lactamase and AME genes in ICU patients.Genetic cluster analysis suggested that clonal propagation result in nosocomial infection of PAE.

Objective To investigate the value of advanced virtual monoenergetic technique(mono-plus) of dual-energy CT (DECT) in improving imaging quality of the lliofemoral arteries.Methods Forty consecutive patients underwent DECT scan.Mono-plus images (40 keV),optimal monoenergy images and mixed energy images were obtained by using Siemens Syngo.via VB10A workstation.CT values,signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR) and subjective image quality were calculated in the abdominal aorta,iliac artery,external iliac artery,femoral artery,profound femoral artery, lateral circumflex femoral artery,transversal branch of lateral circumflex femoral artery,medial circumflex femoral artery and profound branch of medial circumflex femoral artery and compared among the three groups.Results Compared with the optimal monoenergy and mixed energy groups,CT value and CNR of the 9 arteries were significantly higher in the mono-plus group (P<0.003).SNR of the transversal branch of lateral circumflex femoral artery and the profound branch of medial circumflex femoral artery showed no significant difference (P>0.05) among the threegroups,whereas SNR of the arties bigger than the 6th grade branch were significantly lower in the mono-plus group (P<0.05).The subjective image quality in the mono-plus group was significant better than the other two groups(P<0.001).Conclusion 40 keV mono-plus technique in DECT can improve the display of lliofemoral arteries especially the small and distal branches.

Objective: To establish a method of rapid analysis for simultaneous determination of oleanolic acid and ursolic acid in Chinese materia medica (CMM) of Verbena officinalis, Ligustrum lucidum, Prunella vulgaris, Hedyotis diffusa, Patrinia scabiosaefolia, Eriobotrya japonica, Crataegus pinnatifida, Chaenomeles Fructus papaya. Methods: The analyses of preparation were conducted by accelerated solvent extraction (ASE), methanol was used as solvent extraction, and the static extraction time was 6 min. Separation was carried out on Acclaim C30 Thermo column with acetonitrile and 0.2% acetic acid (85:15) as mobile phase, flow rate was 0.3 mL/min, and UV detection wavelength was set at 205 nm. Results: After 10 min sample extraction time, oleanolic acid and ursolic acid reached baseline separation, and no sample matrix was interfered, the linear correlation coefficient was over 0.999, the average recovery between 95.8% and 102.7%; Compared to Chinese Pharmacopoeia 2015, the determination of average mass fraction difference was 3.7% and 5.8%. Conclusion: This method is simple, fast, and suitable for the analysis of these drugs, and it can be used for content determination of oleanolic acid and ursolic acid for eight kinds of CMM.

Objective Inquire into the relationship between diabetic peripheral neuropathy(DPN)pathogenic factor and Antigangliosides antibody(Anti-GS-Ab)in type 2 diabetes mellitus. Methods It was examined by enzyme-linked immunosorbent assays(ELISA)to the levels of serum Anti-gangliosides(Anti-GS)in 2 DM and DPN as well as healthy.Results The positive rate of Anti-GS-IgM,IgG in DPN group were 46.7% and 20.0%.repectivily it was obviously higher than normal group and 2 DM group.Conclusion The relationship between the DPN and Anti-GS-Ab is a close.It show that Anti-GS-Ab play an important role in DPN pathological process.

Objective To study the spectrum of upper gastrointestinal diseases and infection rate of Helicobacter pylori(Hp) in our hospital during the past 35 years.Methods Patients who were 16 or older with duodenal ulcer,gastric ulcer,reflux esophagitis,gastric cancer and esophageal cancer diagnosed by gastroscopy and pathology were retrospectively enrolled in our study from January 1980 to December 2014.Patients with chronic superficial gastritis,chronic atrophic gastritis or Hp infection from January 1989to December 2014 were also included in our study according to the same diagnostic criteria.The incidences of diseases and the infection rates of Hp were studied.Results A total of 213 495 patients underwent gastroscopy in our department during the past 35 years.The overall diagnostic rates of duodenal ulcer,gastric ulcer,reflux esophagitis,gastric cancer and esophageal cancer were 9.87％,3.79％,6.66％,1.59％ and 0.66％ respectively.There were 183 426 patients receiving gastroscopy in our department from January 1989to December 2014.The overall endoscopic diagnosis rates of chronic superficial gastritis and chronic atrophic gastritis were 49.83％ and 22.43％ respectively.The overall infection rate of Hp was 36.18％,which had a declining trend consistent with peptic ulcer (all P =0.000).Yet,the prevalence of reflux esophagitis,chronic superficial gastritis and chronic atrophic gastritis were increasing (all P =0.000).The diagnostic rates of gastric cancer and esophageal cancer were persistent (P =0.266,P =0.156).Conclusions The Hp infection during years has been decreasing,consistent with the declining tendency of peptic ulcer.On the other hand,reflux esophagitis,chronic superficial gastritis and chronic atrophic gastritis show an ascendant trend.The proportion of patients with gastric cancer and esophageal cancer is relatively stable.

Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.

Objective To analyze the effectiveness of antithymocyte globulin(ATG) combined with cyclosporin(CSA)in treatment of pediatric severe aplastic anemia(SAA).Methods Seven children with SAA were treated with ATG and CSA,ATG 4-5 mg/(kg?d),5 days,methyprednisone 2-3 mg/(kg?d) to reduce anaphylaxis,CSA 3-5 mg/(kg?d),assisting with transfusion and G-CSF.Results The complete remission rate was 28.57%(2/7),the partial remission rate was 14.29%(1/7),and the obvious improvement rate was 28.57%(2/7),1 child died.The overall response rate was 71.43%.Transfusion volumes in 4 to 6 months post treatment decreased obviously comparing to the first 3 months(P

Objective To explore the clinical effects of enema combined with massage therapy on jaundice in premature infants.Methods Seventy-five premature infants with jaundice were randomly divided into 3 groups,enema combined message therapy group(group A),abdominal massage therapy group(group B),and double-side phototherapy group(group C).All 3 groups were received the same formula fee-ding,intravenous nutrition and identical drug treatment.Group A was given enema with mixed kaiselu and normal saline together with 60 times clockwise abdominal massage once a day for 2 weeks.Group B only received abdominal massage twice per day for 2 weeks.Transcutaneous bilirubin(TB) indexes of all the premature infants in the 3 groups were detected and transformed into total TB concentrations every morning,through version of MINOLTA JM-102 transcutaneous bilirubin radiometer made in Japan.When TB index counted more than 196.58 ?mol/L,group A and B were given single-side phototherapy for 24 hours.Neither enema nor abdominal massage was given to group C,and double-side phototherapy was applied when TB indexes were above 196.58 ?mol/L.Daily TB indexes,duration of jaundice and phototherapy,time of meconium exhaustion,defecation times in each day,incidence of constipation and feeding intolerance were recorded.Results Duration of jaunhospitalized and phototherapy were significantly shortened in group A compared with those of the other groups.In 34 premature infants who were hospital for at least 2 weeks,TB indexes in group C were lower than those in group B on the 9th day.On the 12th day and the 14th day,TB indexes in group A and C were lower than those in group B(Pa