Matrix Metalloproteinases ; Neovascularization, Pathologic ; Neovascularization, Physiologic

Research on novel pullulanase has major significance on the domestic industrialization of pullulanase and the breakdown of foreign monopoly. A thermophilic bacteria LM 18-11 producing thermostable pullulanase was isolated from Lunma hot springs of Yunnan province. It was identified as Anoxybacillus sp. by 16S rDNA phylogenetic analysis. Full-length pullulanase gene was cloned from Anoxybacillus sp. LM18-11. The optimum temperature of the pullulanase was between 55 and 60 degrees C with a half-life as long as 48 h at 60 degrees C; and its optimum pH was between 5.6 and 6.4. V(max) and K(m) of the pullulanase was measured as 750 U/mg and 1.47 mg/mL, which is the highest specific activity reported so far. The pullulanase crystals structure showed a typical alpha-amylase family structure. The N-terminal has a special substrate binding domain. Activity and substrate binding were decreased when the domain was deleted, the V(max) and K(m) were 324 U/mg and 1.95 mg/mL, respectively. The pullulanase was highly heterologous expressed in Bacillus subtilis by P43 promoter. The extracellular enzyme activity was 42 U/mL, which increased more than 40 times compared to the initial strain. This pullulanase has good application prospects.
Anoxybacillus ; classification ; enzymology ; China ; Glycoside Hydrolases ; metabolism ; Hydrogen-Ion Concentration ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Temperature

OBJECTIVETo study pharmacokinetic effect of Aikeqing Granule (AG) by different medication ways on zidovudine (AZT) in highly active antiretroviral therapy ( HAART) of rats.

METHODSTotally 36 rats were administered with corresponding medications by gastrogavage, group I [HAART: AZT 31.5 mg/kg +3TC 31.5 mg/kg + Efavirenz (EFV) 63.0 mg/kg], group II (HAART+AG525 mg/kg), group III (HAART and AG 525 mg/kg after a 2-h interval). Drug concentrations of AZT were determined by high performance liquid chromatography-mass spectroscopy (HPLC-MS) before HAART, and at 0.5, 1, 2, 3, 4, 6, 8, 10, 12 h after HAART, respectively. Pharmacokinetic parameters [such as t1/2, Tmax, Cmax, AUCo-t, plasma clearance rate (CL)] were calculated by DAS2.0 Software.

RESULTSThe-equation of linear regression of AZT was good, with the precision, coefficient of recovery, and stability definitely confirmed. AUC in group II and III was larger than that of group I. There was no statistical difference in t1/2, Tmax, Cmax, AUC0-12 h, or AUC0-∞ among groups (P > 0.05).

CONCLUSIONAG combined HAART could enhance the Cmax of AZT.

Animals ; Antiretroviral Therapy, Highly Active ; Benzoxazines ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; Mass Spectrometry ; Rats ; Zidovudine ; pharmacokinetics ; pharmacology

Adult ; Female ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Regression Analysis ; Risk Factors ; Stress, Psychological ; psychology ; Surveys and Questionnaires ; Workplace ; psychology

OBJECTIVETo evaluate the effect of penile frenulum lengthening in the treatment of premature ejaculation (PE).

METHODSThirty-four males with PE were enrolled in this study, of whom 8 had received circumcision six months before and 4 had redundant prepuce, all with short frenulum. Those with a history of circumcision underwent reconstruction and lengthening of the frenulum, and those without received frenulum lengthening only.

RESULTSCompared with the baseline, the intravaginal ejaculation latency time (IELT) was significantly increased at 1 month after operation ([1.35 ± 0.49] vs [5.71 ± 2.69] min, t = -9.42, P <0.01), (1.42 ± 0.5) vs (5.31 ± 2.74) min in the patients without circumcision (t = -7.41, P <0.01), (1.12 ± 0.35) vs (7.00 ± 2.20) min in those with circumcision (t = -7.24, P <0.01), and (1.50 ± 0.58) vs (4.75 ± 1.71) min in those with redundant prepuce (t = -3.81, P <0.05). Totally, 94% of the patients were satisfied with their sexual intercourse postoperatively.

CONCLUSIONPenile frenulum plays an important role in penile erection. Reconstruction and/or lengthening of the frenulum can prolong penile erection and IELT in PE patients.

Adult ; Circumcision, Male ; rehabilitation ; Coitus ; Ejaculation ; Foreskin ; surgery ; Humans ; Male ; Penile Erection ; Premature Ejaculation ; surgery ; Reconstructive Surgical Procedures ; methods

ObjectiveTo investigate the clinicopathological features,immunohistochemical phenotype of pancreatic acinar cell carcinoma.MethodsEight cases of pancreatic acinar cell carcinoma admitted to our hospital from January 2001to January 2011were retrospectively analyzed. The clinicopathological characteristics,and immunohistochemical staining for phenotype were analyzed,then the follow-up data were summarized.ResultsAll 8 patients with pancreatic acinar cell carcinoma was male,with a median age of 47 years old.Tumors were located in the pancreatic head in 4 patients,pancreatic body and tail in 4 patients.The average tumor size was 4.5 cm × 4.0 cm × 3.2 cm,the section appeared as gray or gray-red and presented as solid or cystic lesions.Larger tumors were often accompanied by hemorrhage and necrosis.Microscopically,the tumor cells arranged in acinic,cord,trabecular or solid nests.The cytoplasm was abundant and eosinophilic.The nuclear was round,oval,slightly atypia.lmmunohistochemical staining showed diffusely positive for CAM5.2,α-AT,α-ACT and focally positive for CA19-9,CEA,E-cad,β-cat and MUC-1 and only occasionally positive for AFP,NSE,Syn and CgA.Follow-up data showed there was one case of postoperative death due to postoperative pancreatic leakage with abdominal infection.Liver metastasis occurred in 4 cases,among whom,2 cases died.ConclusionsPancreatic acinar cell carcinoma is a rare epithelial malignant tumor of pancreas,with distinct phenotype characteristics.

A transmissible gastroenteritis virus strain was isolated from suspect samples in Sichuan province and identified by ST cell culture, direct fluorescent antibody test (FA), neutralization test (NT), TME examination and some other methods, then it was named SC-Y. The isolated strain could produce obvious cytopathic effects (CPE), The TCID50 was 10(-3.664)/0.05 mL, The neutralization index is 52.5. cDNA fragments covering the complete genome were amplified by the long reverse transcription PCR. The amplified fragments were further cloned and sequenced. The genome of SC-Y strain was assembled by BioEdit. The length of complete genome was 28590 nucletides, and was composed of 7 ORFs, which was flanked by untranslated regions (UTRs) with 315 bases at the 5'-end and 277 bases at the 3'-end. Phylogenetic analysis based on genome suggested that SC-Y might belong to same subgroup with Purdue strain.
Animals ; Base Sequence ; DNA, Viral ; chemistry ; Fluorescent Antibody Technique, Direct ; Microscopy, Electron ; Neutralization Tests ; Phylogeny ; Swine ; Transmissible gastroenteritis virus ; classification ; genetics ; isolation & purification ; ultrastructure

Objective To measure the temperature sensation threshold of trunk skin in healthy adults. Methods The threshold of cold sensation, warm sensation, cold pain sensation and heat pain sensation of trunk skin key points (T3, T7 and T11) were measured with Thermal Sensory Analyzer in 123 healthy adults. Results The thresholds of cold, warm, cold pain and heat pain sensations were obtained. The standard deviation of cold and warm threshold was less than that of heat pain. The range of cold sensation threshold was the largest. The heat pain sensation threshold increased with segmental declining and the sensation threshold increased with age. Conclusion Normal reference value should be established variously with the segment and age. The threshold of cold, warm varies less, while the threshold of cold pain and heat pain varies too much.

In practice of forensic medicine, potential disease can be associated with fatal asphyxia in restraint position. Research has demonstrated that nitric oxide (NO) and nitric oxide synthase (NOS) are plentifully distributed in skeletal muscle, contributing to the regulation of contractile and relaxation. In the current study, respiratory functions, indices of diaphragmatic biomechanical functions ex vivo, as well as NO levels in serum, the expressions of diaphragmatic inducible NOS (iNOS) mRNA, and the effects of L-NNA on contractility of the diaphragm were observed in sepsis induced by cecal ligation and puncture (CLP) under the condition of restraint position. The results showed that in the CLP12-18h rats, respiratory dysfunctions; indices of diaphragmatic biomechanical functions (Pt, +dT/dt(max), -dT/dt(max), CT, Po, force over the full range of the force-frequency relationship and fatigue resistance) declined progressively; the NO level in serum, and iNOS mRNA expression in the diaphragm increased progressively; force increased significantly at all stimulation frequencies after L-NNA pre-incubation. Restraint position 1 h in CLP12 h rats resulted in severe respiratory dysfunctions after relative stable respiratory functions, almost all the indices of diaphragmatic biomechanical functions declined further, whereas little change took place in NO level in serum and diaphragmatic iNOS mRNA expression; and the effects of L-NNA were lack of statistical significance compared with those of CLP12 h, but differed from CLP18 h group. These results suggest that restraint position and sepsis act together in a synergistic manner to aggravate the great reduction of diaphragmatic contractility via, at least in part, the negative modulation of NO, which may contribute to the pathogenesis of positional asphyxia.
Animals ; Asphyxia ; Diaphragm/physiology* ; Muscle Contraction ; Muscle, Skeletal ; Nitric Oxide/metabolism* ; Nitric Oxide Synthase ; Nitric Oxide Synthase Type II ; Rats ; Respiration Disorders ; Restraint, Physical ; Sepsis

BACKGROUNDThe conventional procedure for screening bioactive components from traditional Chinese medicine is time-consuming, expensive and low efficient. Therefore, some alternative strategies are needed urgently. A novel method for screening anti-platelet aggregation components from oleoresins was developed using chicken thrombocyte extract and high performance liquid chromatography.

METHODSThe anti-platelet aggregation components of oleoresins were combined with receptors, channels and enzymes of chicken thrombocytes under physiological environment. Unbound substances were washed away and bound compounds were eluted using specific phosphate buffered solution (PBS). Compounds released from target sites were collected and analyzed by high performance liquid chromatography and LC-MS. The activity of three compounds which were screened from this model was confirmed using platelet aggregation pharmacology in vivo.

RESULTSThere were four typical compounds that bound to the thrombocytes: 6-gingerol, 8-gingerol, 6-shogaol and 10-gingerol, and all had shown anti-platelet aggregation activities. Eight-gingerol displayed the best anti-platelet aggregation effect.

CONCLUSIONSChicken thrombocyte extract can be used to isolate chemicals that are ligands of the receptor or other bio-targets on the platelet. This may therefore be a simple and efficient method to screen for anti-platelet aggregation compounds from traditional Chinese medicine.

Animals ; Catechols ; isolation & purification ; pharmacology ; Chickens ; Chromatography, High Pressure Liquid ; methods ; Fatty Alcohols ; isolation & purification ; pharmacology ; Ginger ; chemistry ; Medicine, Chinese Traditional ; Plant Extracts ; isolation & purification ; pharmacology ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; isolation & purification ; pharmacology ; Rhizome ; chemistry ; T-Lymphocytes ; metabolism