Objective:To express and purify the fusion protein of GST-FILIP-1L and prepare its polyclonal antibody. Methods:The constructed recombinant expression vectors pGEX-4T3-FILIP-1L were transformed into Escherichia Coli BL21. FILIP-1L fusion protein was induced by IPTG and purified by Glutathion Sepharse 4B . The rabbit was immunized by the purified fusion protein,and pro-duced serum with anti-FILIP-1L antibody. The titer of polyclonal antibody was detected by ELISA, the anti-FILIP-1L polyclonal antibody was purified by Active and its combining specificity with FILIP-1L protein was further identified by Western blot. Results:The GST-FILIP-1L fusion protein was highly expressed in E. coli, and its specific polyclonal antibody was obtained after the immunization. The polyclonal antibody purified by Active Ester Agrose was able to combine specially with FILIP-1L protein and transformed FILIP-1L protein in 293 cells and FILIP-1L protein of liver cancer cells, respectively. Conclusion: The GST-FILIP-1L fusion protein was expressed successfully,the anti-GST-FILIP-1L polyclonal antibodies with high titer and specificity are successfully prepared,these antibodies provide an useful experimental tool to research the biological function of the FILIP-1L protein.

Canrenone is an important intermediate for the synthesis of eplerenone,a cardiovascular drug.C_ 11 ?-hydroxylation of canrenone is the key reaction,which can be done by microbial transformation.Rhizopus sp.SIPI-0602,kept in our lab,could high selectively transform canrenone to a compound named SIPI-11.By determining and analyzing the MS,UV,NMR etc.spectra of compound SIPI-11,its chemical structure was elucidated to be 11?-hydroxycanrenone.The study on flask transformation technology showed that the transformation ratio exceeded 90% when the substrate concentration was not more than 6g/L.

Objective:To investigate whether chemokine like factor (CKLF)-like myelin and lympho-cyte and related proteins for vesicle trafficking and membrane link (MARVEL)transmembrane domain-containing protein 2 (CMTM2)is involved in spermatogenesis in varicocele induced sub-fertility rats and to discuss the possible mechanisms.Methods:Forty male SD rats (body weight:220 -330 g,age:6 -7 weeks)were randomly divided into 4 groups:varicocele for 4 weeks,varicocele for 12 weeks,sham operation for 4 weeks and sham operation for 12 weeks,with 10 rats in each group.These rats were intro-duced by partially ligating left kidney veins for the experimental groups,and the sham surgery groups as controls were executed with exactly the same surgery as in the experimental groups except for the ligation. The rats in control and experimental groups for 4 and 12 weeks were killed after laparotomy at the end of 4 and 12 weeks,respectively,the left testes and epididymis were taken out for counting the sperm,ob-serving the seminiferous tubule change and immunochemistry for CMTM2.The changes included sperm density and motility,the outer diameter and inner diameter change and the changes of epithelium and the CMTM2 expression in immunochemistry.Results:Compared with the control groups,the sperm density [(63.9 ±7.1)×106 /mL vs.(74.3 ±5.0)×106 /mL]and motility[(58.7% ±7.9%)vs.(66.1% ± 4.3%)]were reduced slightly in group of varicoele for 4 weeks,respectively (t =1.432,1.563;P =0.076,0.059,respectively ).Varicocele significantly caused a decrease in sperm concentration [(40.5 ±7.2)×106 /mL vs.(71.1 ±4.5)×106 /mL]and motility [(35.2% ±8.5%)vs.(63.4% ± 4.1%)]at 12 weeks,compared with the related sham groups (t =3.754,3.933;P =0.004,0.002, respectively).Additionally,testis CMTM2 exhibited the same disparity,that is,the CMTM2 protein ex-pression in varicocele group was significantly reduced,with the ratio of sham group to varicocele group at the end of 12 weeks 2.3 ±0.4 (t =1.978;P =0.039).In the evaluation of seminiferous tubules diame-ter,the external [(198.2 ±10.2)μm vs.(255.8 ±12.7)μm,t =2.125,P =0.003]and epithelium diameter [(54.1 ±1.5)μm vs.(75.5 ±4.1)μm,t =2.246,P =0.021]were decreased compared with the sham-related groups and previous varicocele groups.In all the varicocele groups,all types of sperm motility decreased compared with the related sham-operated group (P <0.05).Conclusion:This study suggests varicocele has a detrimental effect on CMTM2 levels and decreases spermatogonia cell number,seminiferous tubules diameter,and sperm indices.CMTM2 is associated with sperm changes in rats with varicocele,and further studies are needed to study the mechanism.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Cross-Sectional Studies ; Hepatitis B ; epidemiology ; Humans ; Incidence ; Infant ; Infant, Newborn ; Middle Aged ; Young Adult

Transmembrane protein p185 (the product of Her2/c-erbB-2 gene) is a member of the epidermal growth factor receptor (EGFR) family. Its overexpression was found in about 30% of breast cancer. It is essential to obtain soluble extracellular domain (ECD) of p185, especially disulfide bond rich domains, for identifying the epitopes of anti-p185 antibodies and researching the interrelationship between the antigen and antibody. The disulfide bond rich domain I-II and domain IV of p185 ECD were amplified from plasmid pBabe/erbB-2 by PCR respectively. These two fragments were inserted into pGEX/4T-1 vector, transfected into E. coli Origami B (DE3) pLysS and expressed inductively by low concentration of IPTG and low temperature overnight. After the pressure lysis of cells, the supernatants were analyzed by SDS-PAGE and the result demonstrated that this GST-fusion protein was expressed solubly in the amount of 10-15 mg/L. By the ELISA, Western blot and other immunological assays, the fusion proteins and their GST cut-off derivates both showed binding activities with several anti-p185 antibodies respectively. These results indicated that it was a feasible and effectual method to express disulfide bond rich domain I-II and domain IV of p185 ECD and this method may also be used to express other disulfide bond rich proteins.
Antibodies, Monoclonal ; immunology ; Disulfides ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Receptor, ErbB-2 ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Solubility ; Transfection

OBJECTIVETo study the influence of lipopolysaccharide (LPS)-induced inflammation on the testicular histology and reproductive endocrine function in male rats and investigate the possible mechanism of inflammation affecting male fertility.

METHODSThirty-six male SD rats were randomly divided into a control group (A) and three LPS intervention groups (B, C, and D) to receive saline and LPS (5 mg/kg i. p, once), respectively. The animals in groups B, C, and D were killed by anesthesia at 12, 24, and 72 hours after treatment. Histopathological changes in the left testis of the rats were observed by HE staining and the levels of the reproductive hormones T, FSH, and LH in the serum were determined by ELISA.

RESULTSCompared with group B, group A showed clear structure of seminiferous tubules, orderly arrangement of spermatogenic cells, a slightly decreased number of sperm in some seminiferous tubular lumens, and shed spermatogenic cells in the rat testis tissue; group C exhibited thinner seminiferous epithelia, disordered structure of seminiferous tubules, irregular arrangement of spermatogenic cells, decreased number of mature sperm and obvious shedding of spermatogenic cells in seminiferous tubular lumens; group D manifested similar findings to those of group C, with even more shed spermatogenic cells that blocked the tubular lumens. The levels of serum T, LH, and FSH were (0.490 +/- 0.028) ng/ml, (6.290 +/- 0.515) ng/L, and (1.837 +/- 0.127) IU/L in group A, (0.460 +/- 0.024) ng/ml, (5.881 +/- 0.124) ng/L, and (1.707 +/- 0.098) IU/L in group B, (0.417 +/- 0.021) ng/ml, (5.123 +/- 0.271) ng/L, and (1.620 +/- 0.115) IU/L in group C, and (0.378 +/- 0.021) ng/ml, (4.504 +/- 0.279) ng/L and (1.562 +/- 0.216) IU/L in group D, all decreased in group B as compared with A (P > 0.05). The decreases of T and LH were extremely significant (P < 0.01) and that of FSH was significant in groups C and D (P < 0.05) in comparison with A.

CONCLUSIONLPS-induced inflammation affects the testicular tissue and reproductive endocrine function of male rats, resulting in decreased levels of serum T, LH, and FSH.

Animals ; Endocrine System ; drug effects ; physiology ; Fertility ; drug effects ; physiology ; Follicle Stimulating Hormone ; blood ; Humans ; Lipopolysaccharides ; toxicity ; Luteinizing Hormone ; blood ; Male ; Random Allocation ; Rats ; Reproduction ; Seminiferous Tubules ; drug effects ; pathology ; Spermatocytes ; drug effects ; Testis ; drug effects ; pathology ; Testosterone ; blood

Objective To design an Android-based hand-hold intelligent terminal to meet the requirements for the portability of hyperbaric oxygen therapy information management system. Methods MT6582 processor was used as the primary control unit, which integrated the modules for display, communication, input and output. RAD Studio XE7 was applied to program development, and the main interface had the functions of charging and appointment. Results The hand-hold intelligent terminal gained advantages in stable connection with the server and rapid response, and enhanced the efficiency of hyperbaric oxygen therapy information management system greatly. Conclusion The terminal increases the portability of hyperbaric oxygen therapy information management system, and thus is worthy promoting practically.

The influence of pulsed magnetic stimulation (MS) on the sciatic nerve injury was investigated. Thirty rats were divided into three groups equally: MS group (A), electric stimulation (ES) group (B) and the control group (C). The MS and ES were applied immediately after the first 10 min of the sciatic nerve crush. Sciatic function index (SFI), toe spreading reflex (TSR), muscular weight and volume were measured after the experiment. The TSR of in the groups A and B occurred at 4th day while in the control group it occurs at 10th day. There was statistically significant difference in SFI between groups A and B (P<0.01). The weight and volume of the gastrocnemius muscle were statistically greater in the groups A and B than in the control group (P<0.01). The effect of MS was similar to that of ES. It was suggested that the application of MS immediately after the nerve injury might have an important clinical value as it can accelerate functional recovery and prevent or minimize muscle atrophy. The technique is easily to operate, non-invasion, painless and permits tolerance of high intensity output to be used.

The influence of pulsed magnetic stimulation (MS) on the sciatic nerve injury was investigated. Thirty rats were divided into three groups equally: MS group (A), electric stimulation (ES) group (B) and the control group (C). The MS and ES were applied immediately after the first 10 min of the sciatic nerve crush. Sciatic function index (SFI), toe spreading reflex (TSR), muscular weight and volume were measured after the experiment. The TSR of in the groups A and B occurred at 4th day while in the control group it occurs at 10th day. There was statistically significant difference in SFI between groups A and B (P<0.01). The weight and volume of the gastrocnemius muscle were statistically greater in the groups A and B than in the control group (P<0.01). The effect of MS was similar to that of ES. It was suggested that the application of MS immediately after the nerve injury might have an important clinical value as it can accelerate functional recovery and prevent or minimize muscle atrophy. The technique is easily to operate, non-invasion, painless and permits tolerance of high intensity output to be used.

OBJECTIVETo summarize the experience in nutritional support for the management of duodenocutaneous fistula.

METHODSData of 32 patients with duodenocutaneous fistula in Zhejiang provincial people's hospital from January 1999 to December 2009 were analyzed retrospectively.

RESULTSThe mean duration of nutritional support was 35.6 days (range, 8-82 days). Eight received total parenteral nutrition, 2 total enteral nutrition, and 22 parenteral nutrition combined with enteral nutrition respectively. Succus entericus reinfusion with enteral nutrition was used in 11 cases, glutamine-enriched nutritional support in 28 cases, somatostatin in 12 cases. In these patients, the healing rate was 75.0% after conservative treatment. In the 8 patients who underwent surgery, 6 were cured and 2 died (due to severe abdominal infection and multiple organ failure). A total of 30 patients had the fistulas cured and discharged.

CONCLUSIONSParenteral nutrition combined with enteral nutrition, succus entericus reinfusion combined with enteral nutrition, glutamine-enriched nutritional support and somatostatin are important factors for the healing of duodenocutaneous fistulas.

Adult ; Aged ; Duodenal Diseases ; therapy ; Female ; Humans ; Intestinal Fistula ; therapy ; Male ; Middle Aged ; Nutritional Support ; Retrospective Studies