Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.

Objective To analyze the relationship between different rectal volume,bladder volume and dose of organs at risk (OARs) in intracavitary brachytherapy for cervical cancer.Methods A total of 47 patients with cervical cancer were selected.All of them were treated with high dose rate (HDR) intracavitary brachytherapy with a 600 cGy dose for the dosage point.The effects of different volume of rectum bladder and small intestine for corresponding exposure dose under the standard planning were evaluated using a dose-volume histogram (DVH).According to bladder volume,patients were divided into three groups,＜ 80 cm3 group,80-120 cm3 group and ＞ 120 cm3 group.And according to rectum volume,patients were divided into ＞ 60 cm3 group and ≤ 60 cm3 group.The relationship between the volume and dosage were analyzed.The ANOVA test and t test were used for analyzing D1 cm3,D2 cm3,D30％ and D50％.Results Compared with the group with ＜ 80 cm3 bladder volume,D30％,D50％ value of bladder in groups with 80-120 cm3 and ＞ 120 cm3 of bladder volume increased (F =5.074,5.088,P ＜ 0.05).The difference of D1 cm3 and D2 cm3 value of the small intestine between 80-120 cm3 and ＞ 120 cm3 bladder volume groups were not statistically significant (P ＞ 0.05).D1 cm3 of rectum in groups with ≤ 60 cm3 rectum volume was decreased than that of ＞ 60 cm3 group (t =-2.045,P ＜ 0.05).Conclusions Keeping an appropriatly full bladder and reducing rectal volume in cervical cancers treated with intracavitary brachytherapy can make the exposure dose of bladder,rectum and small intestine relatively small,and reduce the adverse reactions of radiotherapy.

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

OBJECTIVETo investigate the inhibitory effects of bcl-xl antisense oligodeoxynucleotides (ASODN) on growth and gene expression of human nasopharyngeal carcinoma (NPC) in nude mice.

METHODSCNE-2Z cell line was implanted subcutaneously into nude mice. Twenty four h after implantation, bcl-xl ASODN and mismatch control oligonucleotides (SCODN) were injected subcutaneously into nude mice, respectively. The tumor volume and weight were measured twice weekly. The histopathological changes of the tumors were observed by HE staining. RT-PCR and Western-blot were performed for bcl-xl gene expression.

RESULTSGrowth of NPC was significantly inhibited in the ASODN therapy group as compared with that in the control group (P < 0.01). The growth inhibitory rate was 41.7% in the ASODN group and 19.0% in the SCODN group. The expression level of bcl-xl mRNA and protein was decreased in the ASODN group, whereas in the SCODN group there was no significant difference in contrast with saline-treated control group.

CONCLUSIONOur findings suggest that bcl-xl antisense oligodeoxynucleotides results in marked inhibition of NPC growth in nude mice. It may be a novel treatment approach for human nasopharyngeal carcinoma.

Animals ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; genetics ; RNA, Messenger ; analysis ; Transplantation, Heterologous ; bcl-X Protein

OBJECTIVETo study the inhibition of the expression of bcl-xL gene induced by RNA interference in CNE-2Z cell line in addition to the inhibition of its proliferation and apoptotic induction.

METHODSSmall interfering RNAs targeting bcl-xL gene were synthesized by using web design software provided by Amnion and the silencer short interfering RNA (siRNA) construction kit; fluorescein-labeled siRNAs were done by FAM-silencer siRNA labeling kit; siRNAs were transfected into CNE-2Z cells by using lipofectamine 2000 reagent; siRNA transfection efficiencies were analyzed by fluorescent microscopy; down-regulation of bcl-xL was detected by RT-PCR; thiazolyl blue (MTT) assay was used to assess the cell growth; apoptosis of CNE-2Z cells was analyzed by flow cytometry.

RESULTSGreen fluorescence in the cells was seen clearly in FAM-labeled siRNA transfected group under the fluorescent microscope while none in the untransfected group. Different down-regulations of bcl-xL mRNA expression were found in the transfected groups. The expression of bcl-xL mRNA decreased by 10% - 70% in the siRNAs transfected CNE-2Z by RT-PCR scan analysis. The inhibitory rate of cell proliferation depended on time and concentrations to some extent. Different cell apoptosis could be induced by different concentrations of siRNA4.

CONCLUSIONSThe synthesized siRNAs in vitro were able to down-regulate the expression of bcl-xL There were different capabilities of the specific siRNAs down-regulation. The transient transfected bcl-xL siRNA4 could effectively inhibit the growth of the cancer cells and induce theirs apoptosis. It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for anti-nasopharyngeal carcinoma gene therapy.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Messenger ; genetics ; Transfection ; bcl-X Protein ; genetics

Objective To investigate the cell morphology and expression of 5-hydroxytryptamine transporter(5-HTT) in hippocampal CA1 region of depression rats induced by adolescent and post-adult stress,and to observe the inter-vention effect of modified Sini San (MSS). Methods One hundred and thirty-two Wistar rats were randomly divided into blank group,model group,JSS group,and fluoxetine group,33 rats in each group. And then the rats in each group were randomly subdivided into adolescent group, adult group, post-adult group acaccording to the age day, 11 rats in each subgroup. Age day 44,56 and 78 were used as the sampling time points for adolescent group,adult group,post-adult group respectively. Chronic unpredictable mild stress(CUMS)rat model was used. From day 21 to 44 and from day 57 to 78, the rats were modeled and given medication, but from day 44 to 55, the rats were fed normally. The rat general condition and body mass of various groups were observed,the cell morphology of hippocampal CA1 region was observed by hematoxylin-eosin (HE) staining , and the distribution of 5-hydroxytryptamine transporter (5-HTT)positive cells in CA1 region of hippocampus was observed by immunohistochemical staining. Results The general condition of the rats at different age stages in the model group was poor,while that in MSS group and fluoxetine group was improved obviously. The body mass of rats at different age stages in the model group was obviously decreased (P<0.01 compared with the blank group). After adulthood stage,the body mass of rats in model group, MSS group, and fluoxetine group was lower than that of the blank group(P < 0.01), but there was no difference between the 3 groups (P > 0.05). In aspect of cell morphological manifestation in hippocampal CA1 region, rats in the adolescent model group had more deeply-staining atrophy neurons, with unclear hyperchromatic nucleus and cytoplasm. The morphological manifestations in modeled rats at adult stage and post-adulthood stage showed progressive aggravation,manifested as a large amount of neurons stained deeply with unclear nucleus and cytoplasm, and a small amount of glial cells proliferated. Compared with the model group at the same stage,the neuronal atrophy and deeply staining decreased in fluoxetine group and MSS group. The average optical density value of 5-HTT expression in the model group was decreased significantly at the adult stage and after adulthood stage(P<0.05 or P<0.01 compared with the blank group). Compared with the model group, the average optical density value of 5-HTT expression in MSS group after adulthood stage, and in the fluoxetine group at the adult stage and after adulthood stage were increased (P<0.05 or P<0.01). Conclusion Rats suffering CUMS in adolescence presents depressive behavior, and post-adult stimulation can aggravates depression. 5-HTT expression in hippocampus may be an important pathway for MSS to achieve the therapeutic efficacy.

AIMTo investigate the antagonistic effects of the novel compounds on vasoconstriction induced by ET-1 and the effect on the blood pressure of stroke-prone spontaneously hypertensive rats.

METHODSOrgan bath experiment and whole cardiac function experiment were used.

RESULTSThe analogues of o-CPhe-D-Trp-D-Phe(-X)-OH showed good ability against endothelin biological effects. When X was displaced by 3-F, 3-Cl or 4-Cl, the novel compounds inhibit the vascular constriction induced by ET-1 in a concentration-dependent manner, the IC50 +/- L95 were (0.09 +/- 0.05), (0.15 +/- 0.06) or (0.11 +/- 0.03) mumol.L-1 respectively. The blood pressure of stroke-prone spontaneously hypertensive rats was decreased. No significant effect on cardiac function of rats was discovered.

CONCLUSIONThe results demonstrate that among the six kinds of compounds, those with o-CPhe-D-Trp-D-Phe (-X)-OH configuration showed good biological effects.

Animals ; Aorta ; drug effects ; Blood Pressure ; drug effects ; Endothelin Receptor Antagonists ; Endothelins ; pharmacology ; Hypertension ; drug therapy ; physiopathology ; Male ; Molecular Structure ; Peptides ; chemical synthesis ; chemistry ; pharmacology ; therapeutic use ; Rats ; Rats, Inbred SHR ; Rats, Wistar ; Structure-Activity Relationship ; Vasoconstriction ; drug effects

OBJECTIVETo explore brain-protective effect of androgen, its dose-effect relationship and long-term adverse reaction.

METHODSeventy two 3-day-old SD rats were randomized into androgen group (n = 32), HIBD model group (n = 32) and sham operated group (n = 8). The androgen group and HIBD model group were further randomized into 30 mg/kg group, 60 mg/kg group, 120 mg/kg group and 240 mg/kg group, respectively. In androgen group and HIBD group, every rat was given testosterone or peanut oil, one time a day. Three days later, HIBD model was established by occlusion of the left common carotid artery and inhalation of 8% oxygen plus 92% nitrogen for 2.5 hours. Adult rats' ability of learning and memory was determined by water maze test. Escape latencies were recorded and analyzed by statistics. Vaginal cells of all female rats were examined everyday for identifying their estrous cycle. Female rats were allowed to live with normal adult male rats if the female rats were in estrous period. Vaginal cells were examined everyday until sperm was seen, which was the signal of gestation. Pregnancy rate and the number of embryos were recorded and analyzed by statistics. Acropetal coefficient was calculated. The testes and epididymis were taken from adult male rats, histopathological sections were made, and the structure of testis and epididymis were studied under light microscope.

RESULTIn Morris experiment, escape latencies (EL) of HIBD group were much longer than those of sham operation group (27.71 ± 3.19) s, time of first enter target (1(st) ET) was later than that of sham operation group (5.34 ± 0.83) s, times of target cross (TC) was less than that of sham operation group (18.88 ± 1.89) (P < 0.01, P = 0.0005). EL of androgen group (34.89 ± 3.68, 33.71 ± 3.38, 33.84 ± 3.45, 35.43 ± 2.43) were much shorter than that of HIBD group, 1(st) ET (5.39 ± 1.51, 6.28 ± 2.07, 5.09 ± 1.61, 5.85 ± 0.87) was earlier than that of HIBD group, TC (12.75 ± 2.05, 14.88 ± 3.36, 14.88 ± 2.36, 14.38 ± 1.60) was more than that of HIBD group (P < 0.01, P = 0.0001). Among the four doses groups of androgen group, EL, 1(st) ET and TC had no statistical significance (P > 0.05, P = 0.159). There were no statistical significance between male rats of androgen group [Testes acropetal coefficient (0.89 ± 0.07, 0.92 ± 0.08, 0.88 ± 0.11, 0.87 ± 0.09), epididymis acropetal coefficient (0.25 ± 0.02, 0.24 ± 0.05, 0.26 ± 0.04, 0.23 ± 0.05)], HIBD group and sham operation group (P > 0.05, P = 3.207). Among the four doses groups of androgen group had no statistical significance (P > 0.05, P = 6.663). There were no statistical significance between female rats of androgen group (pregnancy rate, 100%; times, 14.52 ± 3.34, 14.69 ± 2.28, 14.98 ± 2.67, 15.38 ± 3.07), HIBD group and sham operation group in pregnancy rate and times.

CONCLUSIONThe intellectual ability of rats decreased after HI. Androgen could reduce the effect of HI on intellectual ability. Androgen had no adverse reaction to the reproductive capacity of adult rats.

Androgens ; adverse effects ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Female ; Hypoxia-Ischemia, Brain ; psychology ; Male ; Maze Learning ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reproduction ; drug effects

The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.
Chemistry, Pharmaceutical ; methods ; Ginsenosides ; chemistry ; Mass Spectrometry ; Panax ; chemistry ; growth & development ; Plant Extracts ; chemistry ; Plant Roots ; chemistry

Objective:To investigate the effect of inhibiting of HOXB7 gene expression on proliferation and apoptosis of colon cancer cells.Methods:The synthetic negative control siRNA (negative control group) and HOXB7-siRNA (HOXB7 transfection group) were transfected into human colon cancer SW480 cells by LipofectamineTM2000 liposome mediated method,untreated cells as blank group.RT-PCR and Western blot were used to detect the mRNA and protein expression of HOXB7 after transfected 48 h respectively;cell proliferation was detected by CCK8 assay after transfected 24,48,72,96 h;cells apoptosis was detected by flow cytometry after transfected 48 h;the expression of apoptosis related proteins Bcl-2,Bax and Notch1 signaling pathway Notch1 and Hes1 were detected by Western blot.Results:The mRNA and protein expression of HOXB7 in HOXB7 transfected group was significantly lower than that in blank group(P<0.05);OD value was no statistical significance in the three groups after transfected 24 h(P>0.05), while after transfected 48,72,96 h,compared with the control group,OD value in HOXB7 group was significantly decreased(P<0.05);compared with the blank group,the apoptosis rate in HOXB7 transfection group increased significantly,and the expression of Bcl-2, Notch1 and Hes 1 proteins was down regulated,and the expression of Bax protein was up-regulated(P<0.05).Conclusion:RNA inter-ference in the expression of HOXB7 gene in colon cancer can inhibit the proliferation of cancer cells and induce apoptosis by inhibiting of the Notch1 signaling pathway.