S801 leukemic cell line established by our department was used as cells for interferon (IFN) induction and stimulating induction. S801 cell culture of concentration of 5-8x10~6 cells/ml was used for IFN induction and IFN titer of 3.55 Lg IU/ml was obtained by classical procedure. The IFN titer obtained S801 cell cultures after pretreatment with Ciwujia eleutherosides B. D and E or polysaccharide (PES) combined with priming procedure,then using Sendai Virus as an inducer was 10-20 times higher than that in cultures without any pretreatment. These Chinese medical herbs may be the ideal IFN helpinducers. This cell line may be applied for IFN production.

OBJECTIVETo investigate the characteristics, diagnosis, and treatment of ovotesticular disorder of sex development (OT-DSD).

METHODSWe retrospectively analyzed 2 cases of OT-DSD treated in our hospital. The patients were 19 and 15 years old, respectively, and both received systematic physical examination and examinations of the karyotype, sex hormone, adrenocorticotropic hormone (ACTH), color Doppler ultrasonography, urethrocystoscopy, and human chorionic gonadotropin (HCG) test. Under the laparoscope, we performed surgical gonad exploration, gonadectomy, and vulvar orthopedics. Intraoperative exploration and pathology confirmed true hermaphroditism in both cases, with sex selection as female. One underwent laparoscopic resection of the ovotestis, and the other removal of the testis with the ovarian tissue reserved.

RESULTSThe patients were followed up for 12 months postoperatively, which found no abnormality in either the vulvas or the genital glands.

CONCLUSIONSurgical exploration of the gonad is the only method for the diagnosis of OT-DSD and sex selection is the key to treatment. Laparoscopic surgical exploration of the gonad and vulvar orthopedics are the first treatment options.

Adolescent ; Chorionic Gonadotropin ; Female ; Gonadal Steroid Hormones ; Humans ; Karyotype ; Laparoscopy ; Male ; Ovary ; Ovotesticular Disorders of Sex Development ; diagnosis ; surgery ; Retrospective Studies ; Testis ; surgery ; Young Adult

An extremely thermoacidophilic isolate K4-1 was obtained from an acidic hot spring in Teng- chong Rehai, Yunnan province. Morphology, growth characteristics, utilization of carbon compounds, en- ergy sources and 16S rRNA gene sequence of K4-1 were studied. Cells of K4-1 are irregular cocci with monotrichous flagella. The strain grew aerobically in either a lithotrophic or a heterotrophic mode. Growth on elemental sulfur occurred through oxidation of sulfur. It grew optimally at 75?C and pH 3.5. On the basis of 16S rRNA gene sequence similarity, strain K4-1 was shown to belong to genus Sulfolobus, being related to the type strains of genus Sulfolobus (86.6%~94.3% similarity), and being most closely related to strain Sulfolobus tengchongensis RT8-4 (98.9% similarity). The GenBank accession number of strain K4-1 16S rRNA gene sequence is EU729124.

Fatigue is a common complication of type 2 diabetes mellitus (T2DM). We examined the relationship between T2DM fatigue and the skeletal muscle 5-hydroxytryptamine (5-HT) system. In animal experiments, a T2DM model was established in mice by feeding a high-fat diet with intraperitoneal injection of streptozotocin. The mice were treated with the 5-HT_2A receptor antagonist sarpogrelate hydrochloride (SH) and the 5-HT synthesis inhibitor carbidopa (CDP) (separately and in combination). In cell culture experiments, C2C12 cells were stimulated with D-glucose, palmitic acid or 5-HT. 5-HT_2AR, 5-HT synthesis and 5-HT degradation were inhibited by SH, CDP, or monoamine oxidase A (MAO-A) inhibitor. The animal experiments were in accordance with the regulations of the Animal Ethics Committee of China Pharmaceutical University. The results showed that 5-HT_2AR, 5-HT synthase and MAO-A were expressed in mouse skeletal muscle and C2C12 cells. The expression of these proteins was significantly up-regulated in T2DM mice or when C2C12 cells were exposed to palmitic acid and D-glucose; palmitic acid was a stronger stimulant of their expression than D-glucose. Rotating rod experiments and biochemical index tests have shown that T2DM fatigue is associated with an increase in skeletal muscle 5-HT_2AR, 5-HT synthesis and 5-HT degradation. 5-HT_2AR mediates the expression of MAO-A and the synthesis of 5-HT, which indirectly regulates the degradation of 5-HT. MAO-A regulates cell inflammation, mitochondrial ROS production and membrane potential depolarization by mediating 5-HT degradation. MAO-A also inhibits the expression of peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1), carnitine palmitoyltransferase-1 (CPT1) and ATP synthase-6 (ATP6), thus inhibiting mitochondrial functions such as fatty acid β oxidation and ATP synthesis. SH and CDP can effectively treat T2DM fatigue, and can also reduce blood glucose and blood lipid, and the combination of SH and CDP has a clear synergistic effect.

OBJECTIVETo establish a rat model of anti-sperm antibody (AsAb)-mediated immune infertility, and investigate the effects of serum AsAb positive on the Fas/Fas-L apoptosis pathway in testis tissue and testicular germ cells of pubertal male rats.

METHODSThirty 5-week-old Wistar male rats were included in this study, 10 killed for preparation of sperm suspension, 10 as normal controls, and the other 10 made models of AsAb-positive immune infertility (experimental group). Four weeks after modeling, the testes of the rats were harvested for observation of the changes in the testis tissue under the light microscope and detection of the expressions of Fas, Fas-L and Caspase-3 proteins by immunohistochemistry.

RESULTSCompared with the control group, the experimental group showed obvious apoptotic changes in the testis tissue and remarkably increased expressions (OD value) of Fas (161.87 +/- 5.37 vs 176.97 +/- 4.58), Fas-L (150.27 +/- 8.65 vs 187.52 +/- 7.76) and Caspase-3 (120.37 +/- 6.76 vs 157.65 +/- 7.38) (P < 0.01).

CONCLUSIONSerum AsAb affected the infertility of pubertal male rats, and its mechanisms might be associated with up-regulated expression of Fas, Fas-L and Caspase-3 proteins in the Fas/Fas-L apoptotic pathway.

Animals ; Apoptosis ; Autoantibodies ; immunology ; Caspase 3 ; metabolism ; Fas Ligand Protein ; metabolism ; Germ Cells ; cytology ; immunology ; metabolism ; Infertility, Male ; Male ; Rats ; Rats, Wistar ; Signal Transduction ; Testis ; cytology ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo explore the regulatory effect of arginine on the secretion of hepatic insulin-like growth factor-1 (IGF-I), and the mechanism of enhancing the immune function by arginine.

METHODSWistar rats were randomly divided into normal control (NC), wound control (WC), and wound with arginine (Arg) groups, with 8 rats in each group. The rats in WC and Arg groups were inflicted with soft tissue trauma on the back. The rats in Arg group were fed a diet supplemented with 5% arginine for one week, while those in NC and WC groups were fed with glycine. The serum contents of arginine, ornithine, growth factor (GH), NO and IGF-I were determined 7 days after feeding. T cell proliferation and IGF-I mRNA expression in hepatic tissue were also measured. Meanwhile, the rat hepatocytes were cultured in serum-free medium containing different concentrations of arginine. The supernatant was collected for the determination of IGF-I level.

RESULTS1). There was no obvious difference of the serum level of arginine and ornithine between NC and WC groups (P > 0.05), but the contents of them were obviously higher in the Arg group compared with other two groups (P < 0.01). 2). No difference in the serum GH level was found among all the groups (P > 0.05), but the serum NO content in WC and Arg groups was significantly lower than that in NC group (P < 0.01), and the serum IGF-I content in WC group decreased obviously compared with that in NC group (P < 0.01). 3). The thymocyte proliferation rate in WC group was also markedly lower than that in NC group (P < 0.01), but that in Arg group was improved compared with WC group (P < 0.01). 4). The expression of hepatic IGF-I mRNA: The relative value of IGF-I mRNA was 1.19 +/- 0.06, 1.08 +/- 0.06 and 1.29 +/- 0.06 in NC, WC and Arg, respectively, while the value in WC was lower than that in NC (P < 0.05) group, and that in Arg group was much higher than that in WC group (P < 0.01). 5). The IGF-I level in the supernatant of cultured hepatocytes: When Arg concentration was 0.0750, 0.7500, 7.5000 mmol/L in the culture medium, the IGF-I level in the supernatant of hepatic cell medi-um was obviously higher than that in the medium without arginine (P < 0.01). Although IGF-I level decreased in the culture medium with arginine in the dose of 37.5000 mmol/L, it was still obviously higher than that in the medium without arginine (P < 0.01).

CONCLUSIONArginine could also produce the immune enhancing effect by stimulating hepatic IGF-I secretion.

Animals ; Arginine ; pharmacology ; Enteral Nutrition ; Insulin-Like Growth Factor I ; metabolism ; Liver ; drug effects ; secretion ; Male ; Rats ; Rats, Wistar ; Soft Tissue Injuries ; metabolism ; therapy

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.

OBJECTIVETo establish a method for the simultaneous determination of 15 pesticides residues in Radix Paeoniae Alba by large volume injection-gas chromatography-mass spectrometry(LVI-GC-MS).

METHODSThe pesticides, including organochlorine pesticides, organophosphorus pesticides and pyrethroid insecticides, were analyzed by LVI-GC-MS using DB-5MS capillary column (30 m X 250 μm, 0.25 μm). The column temperature programming: initial temperature 40 degree for 1 min, with the increasing rate of 20 degree/min to 210 degree for 2 min, then with the increasing rate of 5 degree/min to 280 degree for 22 min. The flow of carrier gas was 1.0 ml/min with the injection volume of 15 μl.

RESULTSThe calibration curves of the pesticides were linear in the specified concentration ranges with correlation coefficients of 0.9937-0.9995. The average recoveries of the pesticides in Radix Paeoniae Alba spiked at two spiked levels ranged from 60.4 % to 106.8 % (for pendimethalin and 4, 4'-DDE those were 53.1 % and 45.2 %) with relative standard deviation(RSD) of 3.6 % to 18.6 % and the detection limits (S/N=3) were in the range of 0.16 μg/kg to 3.59 μg/kg.

CONCLUSIONThe established method for determination of multi-pesticide residue in Radix Paeoniae Alba is rapid, convenient and accurate with high sensitivity and low-cost.

Gas Chromatography-Mass Spectrometry ; methods ; Paeonia ; chemistry ; Pesticide Residues ; analysis

OBJECTIVETo investigate hepatitis E virus (HEV) infection among pigs in Henan province.

METHODSA total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively. The sera positive for HEV antigen were tested for HEV RNA with RT-PCR. The positive products of RT-PCR were cloned and sequenced.

RESULTSThe positive rates of anti-HEV antibody of the groups under 3-month and over 3-month of age were 90.27% and 92.55%, respectively, without statistical difference, while those of HEV antigen were 15.93% and 5.69%, respectively, with significant difference. The positive rates of anti-HEV antibody and HEV antigen were significantly different among different districts. HEV RNA was detectable in 5 of 47 HEV antigen positive samples. The sequence analysis showed that in 4 of 5 specimens the sequence belonged to genotype 4 while in the remaining one the sequence was genotype 1.

CONCLUSIONThe prevalence rate of HEV infection in pigs was high in Henan province and the rate differed in different districts.

Animals ; Antibodies, Viral ; analysis ; immunology ; Antigens, Viral ; analysis ; immunology ; China ; Genotype ; Hepatitis E ; epidemiology ; immunology ; veterinary ; virology ; Hepatitis E virus ; genetics ; immunology ; isolation & purification ; Phylogeny ; RNA, Viral ; analysis ; genetics ; Sequence Analysis, DNA ; Swine ; virology ; Swine Diseases ; epidemiology ; immunology ; virology

In order to build a protein expression system in a cold-adapted bacterium Acinetobacter sp. DWC6, a promoter probe vector was constructed based on the plasmid pBR322. A fragment containing the promoter of the beta-lactamase gene (the ampicillin resistance gene) in pBR322 was eliminated and replaced by a fragment comprizing a kanamycin resistance gene amplified from pJRD215. DNA fragment harboring in the Acinetobacter species specific ori was also inserted into the plasmid pBR322 to construct a promoter probe vector named pBAP1, which could replicate both in E. coli and in Acinetobacter sp. DWC6. The promoter selection library was constructed by randomly inserting genomic DNA fragment of Acinetobacter sp. DWC6 at upstream of reported gene, and target promoters were screened from genomic library on ampicillin selection plates. The function of pBAP1 and isolated promoters were determined by detection of the ampicillin sensitivity and the expression level of beta-lactamase in the host cell.
Acinetobacter ; genetics ; Adaptation, Physiological ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Cold Temperature ; DNA, Bacterial ; genetics ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Models, Genetic ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics ; beta-Lactamases ; genetics ; metabolism