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.

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

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.

Objective To analyze and discuss the content determination of As and Hg from Lilium Lancifolium in Longshan County; To optimize microwave digestion conditions. Methods Automatic microwave digestion appratus was used. Lilium Lancifolium samples from Longshan County were digested in teflon microwave tube with HNO3-H2O2, and As and Hg were measured with atomic fluorescence spectrometry (AFS). Results The content of As was in the range of 0.031–0.507 mg/kg, and the highest content of Hg was 0.024 mg/kg. The regression equation was Y=221.23X+170.72(r=0.995 9),Y=503.52X-682.43,(r=0.999 2).For the production base,the recoveries of As and Hg were 94.32% and 92.48% in the samples, and RSD were 2.14% and 2.70%; for the breeding base, the recoveries of As and Hg were 94.95% and 93.52% in the samples, and RSD were 1.15% and 1.97%. Conclusion The method is simple and reliable, which can be used to the content determination of As and Hg from Lilium Lancifolium, and provide references for the choice of base of production and breeding of Lilium Lancifolium in Longshan County.

Objective To evaluate the clinical efficacy of percutaneous ultra-fine needle CO_2 splenoportography (CO_2-SP).Methods CO_2-SP and 3D-CE-MRA were performed in 36 patients.The imaging quality of the methods was compared by a scoring criterion setup based on the visualization of the trunk,intrahepatic branches of the portal vein and collateral vessels.Results Transient mild abdominal discomfort was presented in 19 patients(52.8% )receiving CO_2-SP.One patient developed snbcapsular splenic hematoma and was discharged with clinical stability several days later after conservative treatment. The imaging quality of the intrahepatic branches of the portal vein with CO_2-SP was much more superior to 3D-CE-MRA (the score was 232 and 198 respectively,t=4.52,P0.05 ).Conclusion Ultrafine needle CO_2-SP is a minimally invasive and safe procedure,able to provide dynamic and clearer imaging of the intrahepatic branches of the portal vein.

Objective To investigate the transfection and expression of p53 genes mediated by liposome and its feasibility in treatment of liver cancer by transcatheter arterial injection on rabbit VX2 hepatocarcinoma model.Methods pCMV-myc-p53 plasmids,LipofectAMINE and p53-LipofectAMINE complex were infused into tumor's feeding artery of rabbit VX2 hepatocarcinoma model,respectively,and then protein of cancer tissue was extracted,followed by measuring gene transfection and expression by western blot and immunohistochemistry,p53-LipofectAMINE complex in different doses were infused into tumor's feeding artery of rabbit VX2 hepatocarcinoma model with the gene transfection and expression detected by the same way.Results Liposome-mediated p53 gene injected through catheter could be successfully transfected and expressed in the cancer tissue of rabbit VX2 hepatocarcinoma model,with transfection efficiency higher than the gene delivery alone.The efficiency and the gene dose has dose-effect relationship.Condusions Treatment of liver cancer by transcatheter arterial injection of p53 genes mediated by liposome is a feasible and effective method,with wide prospect of application.(J Intervent Radiol,2007,16:109-114)

OBJECTIVETo investigate the effects of glutamine enriched enteral feeding on immunoregulation in burn patients.

METHODSTwenty burn patients were randomly divided into enteral nutrition (EN) group and enteral immune nutrition (EIN) group, with 12 patients in each group. Patients in EN group received a standard enteral formula, while those in EIN group received an enteral formula enriched with glutamine after hospital admission. Nutritional support was continued for 10 days. Blood samples were obtained to determine plasma level of total protein (TP), albumin (ALB), prealbumin (PAB) and transferrin (TF) at 1, 4, 7, 10 post-burn days (PBD). At the same time the concentration of immunoglobulin (IgA, IgG and IgM) were determined, the percentage of CD3+, CD4+, CD8+ subpopulations of T lymphocytes, and the ratio of CD4+/CD8+ were determined by FCM.

RESULTS(1) There were no obvious difference of the plasma level of TP, ALB, TF, CD3+, IgM between the two groups at each time-point (P > 0.05). (2) The plasma PAB contents in EIN group were significant higher than that in EN group on 4 PBD [(90 +/- 14 vs 60 +/- 15) mg/L, P < 0.05], 7 PBD [(92 +/- 16 vs 64 +/- 13) mg/L, P < 0.05] and 10 PBD [(106 +/- 21 vs 72 +/-16) mg/L, P < 0.05]. (3) The percentage of CD4+ subpopulation and ratio of CD4+/CD8+ in EIN group were obviously higher than those in EN group on 7 PBD [CD4+ (55 +/- 5 vs 45 +/- 5)%, CD4+/CD8+ (1.92 +/- 0.31 vs 1.53 +/- 0.27)%, P < 0.05] and 10 PBD [CD4+ (56 +/- 5 vs 49 +/- 5)%, CD4+/CD8+ (2.36 +/- 0.36 vs 1.72 +/- 0.42), P < 0.05]. (4) The concentration of IgA and IgG in EIN group were markedly higher than that in EN group on 7 PBD [IgA (2.8 +/- 0.6 vs 2.2 +/- 0.5) g/L, IgG (12.1 +/- 1.3 vs 9.8 +/- 1.2) g/L, P < 0.05] and 10 PBD [IgA (3.1 +/- 0.6 vs 2.5 +/- 0.5) g/L, IgG (14.2 +/- 1.3 vs 10.4 +/- 1.3) g/L, P < 0.05].

CONCLUSIONThese findings suggest that glutamine enriched enteral feeding can improve nutritional status by promoting the synthesis of IgA, IgG, and increasing the PAB concentration, and corrected immunologic dysfunction in burn patients.

Adolescent ; Adult ; Burns ; blood ; immunology ; therapy ; Enteral Nutrition ; Female ; Glutamine ; therapeutic use ; Humans ; Immunoglobulin A ; biosynthesis ; Immunoglobulin G ; biosynthesis ; Male ; Middle Aged ; Prealbumin ; metabolism ; T-Lymphocyte Subsets ; immunology ; Young Adult