With the development of functional genomics, high throughput analysis of genes’ function has been the mainstream of research, and exogenous gene's over expression via Agrobacterium-mediated transformation is the most commonly used method in gene functional analysis.The versatile plant expression vector cassette named pBHT-5 was constructed by the method of site-specific mutagenesis based on pBI121. First of all, the restriction enzyme SfiI recognition site in trfA gene (X00713) which was relevant to plasmid replication and stability was replaced without changing its amino acid composition. And then the SfiIA,SfiIB sites were added between promoter CaMV35s and terminator NOS. The versatile plant expression vector cassette can be directly used to construct plant expression vector containing the full-length genes cloned by Clontech SMARTTM technology, which will raise the efficiency of vector construction. The result will provide basis of new genes’ high throughput screening and functional analysis, then get the new genes functioning in plant osmotic stress resistance.

Objective To evaluate the value of double stapling technique with curved cutter stapler in colorectal anastomosis,especially in low colorectal anastomosis. Methods The clinical data of 168 cases of rectal carcinomas treated with double stapling technique with curved cutter stapler were retrospectively reviewed.The intraoperative condition,postoperative complications and findings during follow up were analysed. Results During the operations,the processes of closure and anastomosis of all the patients were satisfactory,and no operative death occurred.After the operations,4 cases(2.4%) had anastomotic leakage,3 cases(1.8%) had anastomotic bleeding,and 2 cases(1.2%) had rectovaginal fistula.All the complications were cured.There was no anastomotic stricture. Conclusion Double stapling technique with curved cutter stapler may help to accomplish low colorectal anastomosis which is a difficult task for handed suture.

Objective To study the effect of S1P on HLF cell fibrosis and its mechanism. Methods (1) The expression of ECM in HLF cells was analyzed by using Western Blot after treatment by S1P(1 μmol/L), FTY720-P(5μmol/L),ponesimod(5μmol/L)and SEW2871(5μmol/L)24 h;(2)The HLF cells were pre-treated using selective S1PR antagonist W146(1 μmol/L),JTE-013(0.2 μmol/L),and TY-52156(1.25 μmol/L)1 h before incubation by S1P and S1PR agonists 24 h and then the expression of ECM was analyzed;(3)The HLF cells were pre-incubated using JTE-013(0.2μmol/L)and TY-52156(1.25μmol/L)for 1 h and then the expression of ECM was analyzedafter being treated by S1P and S1PR agonists 24 h. Results (1)S1P and selective S1P receptor agonist increased the expression of ECM to various extents;(2)The S1P1R antagonist W146 did not affectthe expression of ECM induced by S1P and S1PR agonists and S1P2R antagonist JTE-013 and S1P3R antagonist TY-52156 both decreased the expression of ECM induced by S1P and S1PR agonists;(3)The expression of ECM induced by S1P and S1PR agonists further decreased using both JTE-013 and TY-52156 but not using ponesimod. Conclusion S1P2R and S1P3R are activated under the influence of S1P so as to increase the synthesis of ECM and promote fibrosis gene expression in HLF cells.

OBJECTIVETo observe the characteristics of sperm single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in infertile men, explore the association of DSB with male infertility, and provide a new observation index and idea for the diagnosis and treatment of the disease.

METHODSThis study involved 60 infertile men (infertility group) and 30 normal healthy males with infertile wives (control group). We comparatively analyzed the seminal parameters of the two groups, determined sperm concentration and viability using the computer aided sperm analysis system, measured the sperm survival rate by hypoosmotic swelling (HOS) test, examined sperm morphology by Diff-Quick staining, and detected sperm DNA damage by two-tail comet assay.

RESULTSNine two-tail comet models were established for detecting sperm DNA integrity. Comparisons between the fertility and control groups showed that the sperm DNA fragmentation index (DFI) was (33.8 ± 13.1) vs (16.3 ± 7.9)% (P < 0.01), the SSB-DFI was (19.2 ± 11.4) vs (14.9 ± 7.6)% (P > 0.05), the SSB-DFI/DFI was (56.8 ± 32.4) vs (91.4 ± 27.8)% (P < 0.01), the DSB-DFI was (23.9 +13.4) vs (6.1 ± 2.7)% (P < 0.01), and the DSB-DFI/DFI was (70.8 ± 19.5) vs (37.4 ± 11.3)% (P < 0.01). The optimal cut-off value of DSB-DFI/DFI in the diagnosis of male infertility was 39.5%, with the AUG, sensitivity, and specificity of 0.969, 98.3%, and 90%; that of DSB-DFI was 15.85%, with the AUC, sensitivity, and specificity of 0.912, 86.7%, and 80%; and that of DFI was 18.65%; with the AUC, sensitivity, and specificity of 0.861, 90%, 70%, respectively. In the infertile men, neither SSB-DFI nor SSB-DFI/DFI exhibited any correlation with semen parameters (P > 0.05); DFI was correlated negatively with the percentage of progressively motile sperm, sperm survival rate, and the percentage of morphologically normal sperm (P < 0.05 or P < 0.01), but not correlated with sperm concentration (P > 0.05); both DSB-DFI and DSB-DFI/DFI showed a negative correlation with sperm concentration, sperm survival rate, and the percentages of progressively motile sperm and morphologically normal sperm (P < 0.05 or P < 0.01).

CONCLUSIONDouble-stranded, rather than single-stranded DNA breaks, may be a factor inducing male infertility. The type of sperm DNA strand damage is of much reference value for the assessment of male fertility.

Case-Control Studies ; Comet Assay ; DNA Breaks, Double-Stranded ; DNA Breaks, Single-Stranded ; DNA Fragmentation ; Fertility ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Semen Analysis ; Sensitivity and Specificity ; Sperm Count ; Spermatozoa ; Staining and Labeling

Aliquat 336 functionalized chelating adsorbent derived from chitosan for enrichment and separation of Y(Ⅲ) were investigated by static adsorption method. The adsorption of Y(Ⅲ) was greatly influenced by the pH of solution, and reached maximum at 20 ℃ using 90 mg/L Y(Ⅲ) at pH 4. 9, and the adsorption of Y(Ⅲ) followed a pseudo-second-order kinetics model and the Langmuir isotherm model. The reduction of Y(Ⅲ) adsorption with the increasing of temperature meant that the adsorption process was exothermic. XPS analysis demonstrated that both cations and anions of the adsorbent were involved in adsorption process, thereby resulting in an improved adsorption of Y(Ⅲ). The adsorbent was thus efficient for enrichment and separation of rare earths from waste rare earth phosphor.

Objective To estimate the efficacy and safety of Voriconazole as antifungal prophylaxis of invasive fungal infection ( IFI) for malignant hematology patients. Methods The randomized controlled trials of Voriconazole treatment in invasive fungal infection for malignant hematology patients (ended in September 2014) were searched from Cochrane library, Medline, Embase, Pubmed, CBM, CNKI, Blood database. The meta analysis were performed by RevMan5.0. Results Ten literatures reported in 1 773 cases, in which there was significantly difference in effective rate between Voriconazole and other antifungal agents such as Amphotericin-B, Itraconazole, Micafungin and Fluconazole(P < 0.000 01); Four literatures indicated that there was significantly difference in adverse event rate between Voriconazole and amphotericin-B (P < 0.00 001); no significantly difference in adverse event rate between Voriconazole and amphotericin-B(P = 0.57); no significantly difference in adverse event rate between Voriconazole and Micafungin (P = 0.69); no significantly difference in adverse event rate between Voriconazole and Fluconazole (P = 0.70); Subgroup analysis indicated that adverse event rate between Voriconazole and Itraconazole is P=0.001, P = 0.17 respectively. Conclusion Voriconazole showed relative high efficient and low toxicity characteristics in treatment of malignant hematology accompanied by invasive fungal infection. But with its widely clinical application, the clinical value of Voriconazole needs to be further tested.

Objective To study the mechanism of killing and apoptosis-inducing effects of Capparis spinosa alkaloid (CSA) on human hepatocarcinoma cell line HepG2. Methods The killing effect of CSA on human hepatocarcinoma cell line HepG2 was studied by MTT method. Morphological observation of HepG2 cells was carried out by fluorescence microscope. Results The CSA had obvious cytotoxicity on the HepG2 in a dose-dependent manner and its IC50 value was 142.82 ?g/mL. The HepG2 cells showed the characteristic morphologic changes of apoptosis by the function of CSA and the apoptosis percentage is higher than that of the natural one. The progress of cells cycle from S phase to G2 phase had been blocked by CSA. The intracellular Ca2+ level had been increased by the function of CSA, which was positively related with drug concentration. Conclusion CSA has obviously killing and apoptosis-inducing effects on human hepatocarcinoma cell line HepG2 and calcium overload might also be invovled in these events.

Objective To explore different doses of calcium 5-formyltetrahydrofolate(CF)for protecting enteral mucosa after chemotherapy of high-dose methotrexate(HD-MTX) in rats.Methods Sixty of 6 weeks old Wistar rats were divided into 5 groups in random,12 rats every group.Group A:control group,normal sodium(NS) intraperitoneal injection only;Group B to E:after HD-MTX intraperitoneal injection(120 mg/kg),1% CF(CF dose amounts to 1% of total MTX dose) for group B,2% CF for group C,8% CF for Group D and empty for group E.For group B、C and D,CF were intramuscular injected after 12 hours of MTX used,q6h?7 times.Rats were killed after 18 hours of the last time of CF.Morphous of jejunum dissection were observed and length of intestinal villus and depth of crypt were mea-sured.Results For group A,jejunum walls were thick and elastic and intestinal villus were close and orderly.Jejunum walls were congestive,swollen and thin,length of intestinal villus and depth of crypt reduced both in group B to E.These were most obvious in group E,and were secondary in group B.Statistical analysis showed that significant difference in effect existed between group B,C,D,E and group A(Pa0.05).Conclusion MTX can damage in intestinal mucosa of rats,CF can reduce this damage,excessive low doses of CF can't play this role.

Animals ; Disease Progression ; Kidney Diseases ; physiopathology ; Kidney Glomerulus ; drug effects ; pathology ; Rats ; Renal Agents ; pharmacology ; Renin-Angiotensin System ; drug effects ; Sclerosis

OBJECTIVETo investigate the treatment of upper lip atrophy resulted by previous therapy.

METHODSFrom Mar. 2008 to Mar. 2012, 4 cases with upper lip atrophy resulted by radiotherapy and sclerosing agent injection were treated with lower orbicularis oris muscle flap wrapped by acellular dermis. The thickness and height of upper lip were increased to improve the lip atrophy.

RESULTSPrimary wound healing was achieved in all the 4 cases. All the patients were followed up for 3 years with obvious improvement and inconspicuous scar.

CONCLUSIONThe volume of lower lip in children is not sufficient as donor site. The lower orbicularis oris muscle flap wrapped by acellular dermis can effectively improve the lip thickness and vermilion portion of upper lip.

Acellular Dermis ; Atrophy ; surgery ; Child ; Female ; Humans ; Lip Diseases ; surgery ; Male ; Mouth Mucosa ; transplantation ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps