The protective roles of tiangui gengnian soft capsule on brain in aged female rats and its mechanism / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the brain-protective and anti-aging effects and the mechanism of action of Tiangui Gengnian Soft Capsule (TGSC), a Chinese herbal preparation composed of sea buckthorn fatty acids.</p><p><b>METHODS</b>Sixteen-month-old female rats were administered via gastric perfusion with low, medium and high doses (0.72 g/kg, 1.80 g/kg and 4.5 g/kg) of TGSC for 180 days. The 3-month and 22-month old rats were taken as controls. Expression of estrogen receptor beta(ERbeta), tyrosine hydroxylase (TH), granulocyte colony-stimulating factor (G-CSF) and vascular endothelial growth factor (VEGF) in the brain of rats were observed and analyzed using immunohistochemical technique, and changes in the number of neurons and glial cells were counted with Nissl's staining.</p><p><b>RESULTS</b>Immunohistochemistry showed the number of ERbeta- and TH-positive neurons in the hypothalamus was significantly higher (P < 0.01 and P < 0.05), G-CSF-positive astrocyte stain was much weaker (P < 0.01) and the expression of VEGF-positive cerebral vessel was stronger (P < 0.05) in the TGSC treated groups than those in the control. Nissl's staining showed no significant difference in the number of neurons and glial cells among groups.</p><p><b>CONCLUSION</b>TGSC could up-regulate the expressions of ERbeta, TH and VEGF in the brain of aged female rats, modulate the responsibility of astrocyte and its G-CSF expression, suggesting that TGSC may have certain neuro-protective and anti-aging effects.</p>

The reproductive system impairment of adult male rats induced by cocaine / 法医学杂志

OBJECTIVE@#To investigate the reproductive system impairment induced by cocaine in adult male rats and the possible underlying mechanism.@*METHODS@#Thirty adult male rats were randomly divided into experimental and control groups, with 15 rats in each group. Rats of the experimental group were injected cocaine hydrochloride (15 mg/kg body weight) subcutaneously daily for four weeks. The weight of body and testis, as well as the level of serum hormone of the rats were examined. In addition, the apoptosis rate of testicular tissue by TUNEL and the expression of Fas gene in testicular tissue were examined by immunohistochemistry.@*RESULTS@#Compared with the control, the weight of testis in the cocaine exposed group decreased significantly (P<0.05), and the serum testosterone level decreased significantly (P<0.05). Moreover, both the apoptosis rate and the expression of Fas gene increased in the testicular tissue of rats in the cocaine exposed group in comparison to the control group (P<0.05). The apoptosis rate was significantly correlated with the expression of Fas gene (r=0.9012, P<0.05).@*CONCLUSION@#Cocaine may cause reproductive system injury in adult male rats, and Fas-mediated apoptosis may be one of the functional mechanisms involved in the reproductive system injuried by cocaine.

Gender-dependent difference of NF-kappaB expression in the hippocampus of prenatally stressed offspring rats / 生理学报

In this study, immunohistochemistry and Western blot were used to determine whether the expression of NF-kappaB in the hippocampus of prenatally stressed offspring rats is gender-dependent. The results were as follows: In the female offspring rats, the expressions of p65 in the hippocampal dentate gyrus in mid-term stress (MS) and late-term stress (LS) groups were significantly less than that in the control group (P<0.01). There was a significant difference between MS and LS groups (P<0.01). The expressions of p50 in all regions of hippocampus in MS and LS groups were significantly more than that in the control group (P<0.01). A significant difference was also present between MS and LS groups (P<0.01). In the male offspring rats, the expressions of p65 in the hippocampal dentate gyrus in MS and LS groups were evidently more than that in the control group (P<0.01). There was a significant difference between MS and LS groups (P<0.01). The expressions of p50 in all regions of hippocampus in MS and LS groups were significantly less than that in the control group (P<0.05, P<0.01). There was also a significant difference in p65 expression between MS and LS groups (P<0.01). In addition, in the control group the expressions of p65 in the hippocampal dentate gyrus of female offspring rats were significantly more than that of male ones (P<0.01). However, in LS group the expressions of p65 in the hippocampal dentate gyrus of female offspring rats were significantly less than that of male ones (P<0.01). Moreover, there was no significant difference in p65 expression between female and male offspring rats in MS group. In the control group the gender difference in the expression of p50 was only observed in hippocampal CA1 (P<0.01). The expressions of p50 in all regions of hippocampus of female offspring rats were significantly more than that of male ones in LS group (P<0.01). There was no significant difference in p50 expression between female and male offspring rats in MS group. The results of Western blot were similar to those of immunohistochemical study. These results indicate that prenatal stress in different gestational periods significantly affects the expressions of p65 and p50 in hippocampus, and this effect is gender-dependent. This may be one of the mechanisms underlying the gender difference in the ability of learning and memory of the prenatally stressed offspring rats.

A method with TRIzol~ reagent and liquid nitrogen to extract high-quality RNA from rat pancreas / 西安交通大学学报(医学版)

Objective To establish a quick,economical and reproducible method for high-quality RNA extraction from pancreas.Methods We utilized TRIzol Reagent and liquid nitrogen to isolate total RNA from the rat pancreas.The RNA quality was determined by detection of its content and optic density(A) at 260/280nm,and electrophoresis in 1% non-denatured agarose gel.Then reverse transcription-polymerase chain reaction(RT-PCR) was performed to detect expression of the pancreas-specific genes.Results The content of the total RNA extracted from the rat pancreas reached 3-6?g/mg pancreatic tissues,and A260/280 ratio was 1.75-1.89.Electrophoresis of the total RNA showed 28S and 18S rRNA bands with clear smear between them.The RT-PCR products of pancreas-specific genes including insulin 1,glucagon,?-amylase and housekeeping gene ?-actin all exhibited clear bands on 1% agarose gel,which were located in the expected positions,respectively.Conclusion These results suggest that we have successfully isolated the high-quality and intact RNA from the rat pancreas with TRIzol Reagent and liquid nitrogen.The extracted total RNA can be used in RT-PCR for pancreatic gene expression.

Detection of HPV 58 and cloning of its E7 gene in cervical cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect HPV 58, a common type of human papillomavirus (HPV), clone and express its E7 gene from biopsy specimens of cervical cancer.</p><p><b>METHODS</b>HPV 58 from 58 biopsy tissues of cervical cancer was detected by GP5+/GP6+ PCR followed by template-directed dye-terminator incorporation assay with fluorescence polarization detection (TDI-FP). HPV 58 E7 gene was amplified from one HPV 58-positive sample, and then cloned into pGEM-T Easy vector. The recombinant plasmid, HPV58E7-pGEM-T was confirmed by sequencing. Subsequently, E7 gene was cloned into prokaryotic expression vector pRSET-A. The constructed pRSET-58E7 plasmids were transfected into BL21(DE3) cells, and induced to express 58 E7 protein by IPTG.</p><p><b>RESULTS</b>Among the 58 biopsy tissues of cervical cancer, 10 were HPV 58-positive, accounting for 19.2% of 52 HPV-positive cases. HPV 58 E7 gene was amplified from one HPV 58-positive sample. The constructed plasmids were identified containing HPV58 E7 gene by restriction enzyme analysis and sequencing. SDS-PAGE analysis showed that HPV58 E7 His6 fusion protein of M(r) 16 x 10(3) was expressed by pRSET-58E7 after induction by IPTG. The fusion protein accounted for 30% of total bacterial proteins.</p><p><b>CONCLUSION</b>HPV 58 is not uncommon in Chinese women with cervical cancer in Shaanxi province. Constructed HPV58 E7 recombinant plasmids can be effectively expressed in E.coli, which may provide a tool in diagnosis and vaccine design for HPV of HPV58-associated tumors.</p>