OBJECTIVETo explore the methods of isolation, cultivation, purification, identification of the fetal mice testis Leydig cell and to observe its biological characteristics in vitro.

METHODSLeydig cells were isolated by 0.03% collagenase (type I) from fetal mice testis and cultured in DMEM/F12 medium. The identity and purity of Leydig cell were assessed by 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD). Cell viability was measured by trypan blue. Testosterone level in the medium of cultured Leydig cells was measured in various culture phases and cell density by radioimmunoassay.

RESULTSThe purity of Leydig cell was (45.10 +/- 1.66)% before culture, and (81.17 +/- 2. 32)% 72 h after culture. The level of testosterone secreted by Leydig cells could be detected in the medium and its level was associated with the density and time of cultured Leydig cells. The secretion capacity of testosterone by single Leydig cell decreased gradually during the culturing period.

CONCLUSIONThe fetal Leydig cells isolated from fetal mice testis have high purity. It can be cultured and kept the secretion ability of testosterone for a few days in vitro. This system can provide a valuable model for further study on the cellular function of the Leydig cells of fetal mice.

Animals ; Cell Separation ; Cells, Cultured ; Leydig Cells ; cytology ; physiology ; Male ; Mice ; Mice, Inbred Strains ; Testis ; embryology ; Testosterone ; secretion

Male sterility affects human reproduction seriously. Modern medicine has not yet fully understood the reasons for abnormal sperm, so clinical treatment is mainly based on experience, but the effect is inaccurate. According to "kidney controls reproduction" and "chronic illness causes blood stasis" theory, Professor JIN Bao-fang treats abnormol speerm sterility by tonifying kidney and activating blood circulation to remove obstruction with modified Yangjing Decoction, which has achieved good efficacy.

OBJECTIVETo Verify the safety and reliability of one-stage repair of complete cleft Lip and palate in infancy and to obtain the primary result.

METHODSThe simultaneous repair of complete cleft Lip and palate in infants 3 to 12 months of age were performed in 271 cases. The deformities include 185 cases of typical complete unilateral clefts and 75 cases of complete bilateral clefts, and other 11 atypical cleft infants. The preoperative orthopedic treatment for wide alveolar cleft was undertaken in 24 infants and the lip appearance and speech outcome were evaluated in 116 children by 1 to 4 years' postoperative follow-up.

RESULTSAll infants, except for dyspnea in 2 babies, palatal fistula formation in 6 cases and temporary wound hemorrhage in 5 infants, were recovered without complications. After orthopedic treatment, the width of the alveolar cleft was reduced 6.1 mm in average. The evaluation showed that 93.1% of children had got good or excellent lip appearance. And the acceptable or excellent speech was found in 94.8% children.

CONCLUSIONSSimultaneous repair of complete cleft lip and palate in infancy is safety and reliable. The preoperative orthopedic procedure is able to reduce the wide alveolar cleft and to achieve alignment of alveolar segments. The acceptable and or excellent lip appearance and speech function could be obtained in this one-stage operative procedure in infants.

Cleft Lip ; surgery ; Cleft Palate ; surgery ; Female ; Humans ; Infant ; Male ; Treatment Outcome

Objective To investigate the effect of leptin on the tolerance of cultured rat brain astrocytes to ischemia and hypoxia.Methods The brain astrocytes isolated from neonatal SD rats,after purification and identification,were incubated in serum-and glucose-flee medium in the presence of 5%CO2+95%N2 for 90 min to induce isehemic and hypoxic injury. RT-PCR was performed to detect the expressions of the leptin receptors Ob-Ra and Ob-Rb in the cells, and colorimetry was used to measure the content of malonaldehyde(MDA) and lactate dehydrogenase(LDH) activity in the cell supematant.The expression level ofglial fibrillary acidicprotein(GFAP)in the cells was detected with fluorescence immunocytochemistry.Results Ischemic and hypoxic exposure of the cells induced obvious cell necrosis.Compared with the cells without the exposure,significantly decreased Ob-Rb expression(0.52±0.01 vs 1.32±0.01,P<0.05)and increased MDA,LDH and GFAP levels(709.68±47.16 vs 516.13±29.08,3.94±0.36 vs 1.81±0.21,and 0.122±0.016 vs 0.057±0.006,respectively,P<0.05) occurred after the exposure,whereas the expression level of Ob-Ra underwent no significant changes(3.87±0.13 vs 3.96±0.24,P>0.05). Compared with the exposed cells,the leptin-treated cells showed a significant reduction in MDA levels(3.94±0.36 vs 3.19±0.25,P<0.05) with significantly increased GFAP expression(0.057±0.006 vs 0.109±0.008, P<0.05)after the exposure, and the cells maintained basically intact cell morphology.Conclusion With neuroprotective effects against ischemic neuronal injuries,leptin canimprove the tolerance of rat brain astrocytes to ischemia and hypoxia.

To study the phenolic constituents from the dry stem of Juncus effusus L. , the constituents were isolated by normal-phase and reverse-phase silica gel column chromatography from the EtOAc extract. Their structures were elucidated by spectral analysis. Six phenolic constituents were purified and identified as 7-carboxy-2-hydroxy-1-methyl-5-vinyl-9,10-dihydrophenanthrene (1) , 2,3-isopylidene-1-O-ferulic acid glyceride ( 2 ) , ( 2S )-2, 3-isopylidene-1-0-p-coumaroyl glyceride (3 ) , dehydroeffusal ( 4 ) , p-hydroxybenzaldehyde (5) and luteolin-5,3'-dimethyl ether (6). Compounds 1 and 2 are new compounds. Compounds 5 and 6 were isolated from Juncaceae plant for the first time. 13C NMR data of compound 6 were reported for the first time.
Anthracenes ; chemistry ; isolation & purification ; Benzaldehydes ; chemistry ; isolation & purification ; Coumaric Acids ; chemistry ; isolation & purification ; Flavones ; chemistry ; isolation & purification ; Magnoliopsida ; chemistry ; Molecular Conformation ; Molecular Structure ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

OBJECTIVETo express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.

METHODSHA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.

RESULTSWe successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.

CONCLUSIONWe expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.

Animals ; Blotting, Western ; Cell Line ; Drosophila ; Female ; Gene Expression ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; chemistry ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; metabolism ; Influenza, Human ; virology ; Mice ; Mice, Inbred BALB C ; Solubility

OBJECTIVETo study the regulatory effect of Bushenfang on the serum testosterone (T) level of naturally aging rats and its mechanism, in order to provide a theoretical and experimental basis for the clinical treatment of late onset hypogonadism (LOH) in males.

METHODSThirty-two 18-month-old male SD rats were randomly divided into four groups of equal number, naturally aging model and low-, medium- and high-dose Bushenfang groups, and another eight 4-month-old rats were taken as normal controls. The rats of the aging model and normal control groups were treated with normal saline, while those of the low-, medium- and high-dose Bushenfang groups received intragastrically Bushenfang at 3.25, 7.50 and 15.00 g/kg, respectively, all for 3 weeks. Then the rats were sacrificed, the histomorphologic changes of the testis observed by HE staining, the serum T level measured by radioimmunoassay, and the expressions of the StAR protein, P450scc and 3beta-HSD I determined by RT-PCR.

RESULTSThe number of Leydig cells was obviously increased after Bushenfang treatment. The levels of serum T were significantly higher in the low-, medium- and high-dose Bushenfang groups ([6.74 +/- 1.56] nmol/L, [8.50 +/- 1.99] nmol/L and [12.41 +/- 2.91] nmol/L) than in the model group ([3.48 +/- 0.75] nmol/L) (P < 0.05). The three Bushenfang groups also showed a remarkable elevation in the mRNA expressions of StAR (0.74 +/- 0.29, 0.83 +/- 0.32 and 1.35 +/- 0.50), P450scc (0.72 +/- 0.36, 1.023 +/- 0.30 and 1.41 +/- 0.37) and 3beta-HSD I (0.58 +/- 0.14, 0.72 +/- 0.07 and 0.85 +/- 0.18), as compared with the models (StAR: 0.44 +/- 0.09; P450scc: 0.33 +/- 0.05; 3beta-HSD I: 0.34 +/- 0.02), with significant differences in the StAR expression between the high-dose Bushenfang and the model groups, as well as in P450scc and 3beta-HSD I expressions between the medium- and high-dose Bushenfang and the model groups (P < 0.05).

CONCLUSIONBushenfang could improve the pathological status of testicular injury and increase the expression of testosterone synthetase, which might be the mechanism behind its regulatory effect on the serum T level of aging rats.

Aging ; drug effects ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hypogonadism ; drug therapy ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; drug effects ; Testosterone ; metabolism

This study was aimed to investigate the polymorphism of D2S1338 and D19S433 loci in Southern Chinese Han nationality population, and to evaluate its forensic application. The DNA was extracted by chromatography on che-lex-100. Fifteen STR loci (D2S1338, D19S433 etc.) were amplified by Identifiler kit and analyzed by 3100 genetic analyzer. The softwares of analysis were GeneScan 3.0 and Genetype 3.7. The results indicated that the allele frequencies of H, DP, PIC, PE of D2S1338 and D19S433 loci were obtained. The allele frequency of samples were in Hardy-Weinberg equilibrium by chi2 test. No mutation was found on D2S1338 and D19S433 loci in 370 cases of meiosis. It is concluded that the D2S1338 and D19S433 loci are high polymorphic and their mutation rates in Southern Chinese Han nationality population are low. Their good distribution is suitable for forensic individual identification and paternity testing.
Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; DNA Fingerprinting ; methods ; Forensic Medicine ; methods ; Gene Frequency ; Genetic Variation ; Humans ; Polymorphism, Genetic ; Tandem Repeat Sequences

To study the correlation between acute lymphoblastic leukemia (ALL) and HLA-A, B and DRB1 gene in southern Chinese Han population and to investigate the susceptible HLA gene to ALL, a total of 4707 healthy volunteer bone marrow donors from southern Chinese Han population were used as a control group, 201 patients diagnosed as patient group from southern Han individuals were genotyped at HLA-A, B and DRB1 loci by PCR-SSP, PCR-SSOP and SBT. HLA allele frequency and its distribution of ALL patient group were compared with the control group by using chi(2) test, and calculated the statistic value of relative risk (RR), pathogenicity score (EF) and preventive score (PF). The results showed that in comparison with the control group, the gene frequence of HLA-A26, B56 and DR9 increased significantly, but the gene frequence of HLA-A30, A33 and B58 allele frequency decreased significantly for patients with ALL. It is concluded that HLA-A26, B56 and DR9 gene have a high correlation with ALL and seem to contribute the genetic susceptibility to ALL in southern Chinese Han populations. However, HLA-A30, A33 and B58 gene seem to have protective role for southern Han individuals suffered from ALL.
Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Chi-Square Distribution ; Child ; Child, Preschool ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; ethnology ; genetics

Objective To compare the clinical efficacy among moxibustion plus pelvic floor muscle exercises, pelvic floor muscle exercises, and Western medication in treating postpartum stress urinary incontinence. Method Fifty-eight eligible patients were divided by the random number table into an integration group (19 cases), an exercise group (19 cases) and a Western medication group (20 cases). The integration group was intervened by pelvic floor muscle exercises plus moxibustion, 3 times of exercises a day, 5 min each time, and moxibustion at Zusanli (ST36), Sanyinjiao (SP6), Guanyuan (CV4), Zhongji (CV3) and Qihai (CV6), once a day, 30 min each session. The exercise group was intervened by muscle force exercises to train the pelvic floor muscle, 3 times a day, 5 min each time. The Western medication group was intervened by oral administration of Duloxetine hydrochloride, 80 mg a day (40 mg per dose, twice a day). For the above methods, 6 d was taken as a course of treatment. Four treatment courses were observed, with a 1-month follow-up study. The 1-hour pad test was used to record the volume of urine leak. Before and after the treatment, as well as in the 1-month follow-up, the International Consultation on Incontinence Questionnaire-Urinary Incontinence-Short Form (ICIQ-UI-SF) score was evaluated and compared among the groups, and the short-term and long-term therapeutic efficacies were also compared. Result The short-term and long-term therapeutic efficacies of the integration group were superior to those of the exercise group and Western medication group (P<0.05). The integration group showed significant improvement after the treatment and in the follow-up study compared to the pre-treatment state (P<0.01); after the intervention, the integration group was better than the exercise group and Western medication group in comparing the urine leak volume by the 1 h pad test and ICIQ-UI-SF score (P<0.01,P<0.05). Conclusion Moxibustion plus pelvic floor muscle exercises can improve the short-term quality of life of patients with postpartum stress urinary incontinence. This method can produce a better effect in improving the quality of life compared to pelvic floor muscle exercises alone.