Objectives:To establish a novel alopecia animal model in multi-strain mice.Methods:BARB/c mice,129 mice,C57mice and C3H/HeJ mice were purchased at the age of 6 to 8 weeks. 30 mice of each kind were divided into control and experimental groups randomly. The control group were treated with vehicle alone. The experimental groups were treated with imiquimod cream with cotton swaps for back skin 3 times a week. The existence of TLR7 on skin was examined by immunohistochemistry(IHC). Results:Alopecia lesions were observed in all the mice in experimental group,but mice in control group could not induce by imiquimod. The aggregation of TLR7were observed in hair follicle. Conclusions: Imiquimod can stably induce non-inflammatory alopecia in BARB/c mice,129 mice,C57mice and C3H/HeJ mice by activating TLR7.

Parkinson disease(PD) is a common disease in the middle-aged and elderly population,which may affect their quality of life.Nowthere are no effective curing methods,however,the early diagnosis is important for heightening the therapeutic effect.In nuclear medicine,people have achieved great progress in utilizing SPECT and PET, which will become one or several common diagnosis means. This owes to the successful development of the imaging agent with superior behavior.

Objective: To investigate the effect of deguelin on mitochondrial permeability transition pore of human breast cancer cell line MDA-MB-231. Methods: The inhibitory effect of deguelin on cell proliferation was determined by MTT assay; cell apoptosis rate was analyzed by flow cytometry with AnnexinV FITC/PI double staining; MDA-MB-231 cells were stained by Rhodaminel23 to detect the changes of mitochondrial transmembrane potential by FCM; alteration of protein of Cyt c outside of mitochondria was detected by Western blotting analysis; caspase-3 activity was assessed by colorimetric assay; and MDA-MB-231 cells were stained by Fluo-3/AM to detect changes of intracellular Ca 2+ concentration by FCM. The expression of Bcl-2 and Bax was examined by RT-PCR and Western blotting analysis. Results: Deguelin significantly inhibited the growth of MDA-MB-231 cells in a time- and dose-dependent manner ( P < 0.05). After treatment with deguelin, mitochondrial transmembrane potential was decreased, the expression of Cyt c outside of mitochondria was increased, and caspase-3 activity was significantly increased compared with negative control group(P<0.01). FCM analysis showed that the apoptotic rate of MDA-MB-231 cells and intracellular Ca2+ concentration increased gradually with the increase of deguelin concentration. RT-PCR and Western blotting analysis showed that the expression of Bcl-2 mRNA and protein was down-regulated and that of Bax was up-regulated after deguelin treatment. Conclusion: Deguelin can inhibit proliferation and induce apoptosis of MDA-MB-231 cells, and the induction of apoptosis might be related to increased intracellular Ca2+ concentration and changes of mitochondrial permeability transition pore induced by altered Bcl-2, Bax expression.

Streptococcus gordonii is a nonpathogenic gram-positive commensal bacterium and component of the normal microbial flora of the human oral cavity. It is suitable to be as a mucosa vaccine vector due to its special bionomics. knowing the bionomics of Streptococcus gordonii,the general expressing system,and the application of it in mucosa vaccine,will provid important reference for the further development of its mucosa vaccine.

To construct expressing vector carrying esat6 gene and express this protein in Streptococcus gor-donii GP251. esat6 gene was amplified by PCR with specific primer from genome of Mycobacterium tuber-culosis (MTB)H37Rv. Inserted esat6 into the pMD18-T vector by T/A clone to get recombinant vector pMD18-esat6. Then digested pMD18-esat6 with restriction enzyme, esat6 was cloned to vector PSMB104 and expressed in Streptococcus gordonii GP251. The expression of esat6 protein was detected by Tricine-SDS-PAGE and Western-blot, ELISA technique was also used to detect its secretory volume. Re-striction endonuclease, PCR, Tric ine-SDS-PAGE and Western-blot confirmed that esat6 gene was cloned into expressing vector successfully, and a 10 kD protein secreted in Streptococcus gordonii GP251, this pro-tein has a good immunogenicity. The expression vector of esat6 gene was constructed, and esat6 protein ex-pressed in Streptococcus gordonii1 successfully, it will be benefit for future study.

Genetic Engineering ; Humans ; Rabies ; virology ; Rabies virus ; genetics ; pathogenicity ; Viral Proteins ; genetics ; Virulence

Objective To observe the clinical efficacy of thread embedding plus Chinese medication in treating primary dysmenorrhea due to blood stagnation caused by cold accumulation. Methods Totally 136 patients with primary dysmenorrhea due to blood stagnation caused by cold accumulation were randomized into a treatment group and a control group, 68 in each group. The treatment group was intervened by thread embedding at Shiqizhui (EX-B 8) and bilateral Ciliao (BL 32) plus Chinese medication;while the control group was by Chinese medication alone. The dysmenorrhea symptom score and Visual Analogue Scale (VAS) were observed before and after intervention, and the clinical efficacies were compared. Results The total effective rate and recovery-markedly effective rate were respectively 93.8%and 78.1%in the treatment group, versus 85.2%and 50.0%in the control group, and there was a significant difference in comparing the recovery-markedly effective rate (P<0.05). After intervention, the dysmenorrhea symptom score and VAS score were significantly changed in both groups (P<0.01);there were significant differences in comparing the symptom score and VAS score between the two groups (P<0.05). The half-a-year follow-up study showed that the total effective rate and recovery-markedly effective rate were respectively 90.6%and 68.8%in the treatment group, versus 70.6%and 38.2% in the control group, and the differences were statistically significant (P<0.05, P<0.01). Conclusions Thread embedding plus Chinese medication is an effective approach in treating dysmenorrhea due to blood stagnation caused by cold accumulation, and it’s easy-to-operate and causes less pain.

Objective To construct the eukaryotic expression vector coding HCV gene E2 fused with His-Tag, and to express fused protein in CHO cells for investigating the function of HCV envelope protein E2. Methods The gene encoding HCV envelope protein E2 was amplified from pBRTM/HCV1-3011, a plasmid containing the cDNA of HCV's ORF, by polymerase chain reaction (PCR) method and cloned into the vector pET28(a) containing His-Tag to obtain the fused HCV envelope protein E2 gene fused with His-Tag. The fused gene was cloned into pcDNA3.1 to construct the recombinant plasmid pcDNA3.1-His-E2, which will express the E2 protein, fused with His tag. This recombinant plasmid was transfected into CHO cells by Lipofactamine 2000 reagent. The fused protein was identified by indirect immunofluorescence (IIF) and Western-blot (WB) methods. Result The positive results were obtained when the fused protein of HCV E2 with His-Tag were identified by IIF and WB methods. Conclusion The eukaryotic expression vector pcDNA3.1-His-E2 was constructed successfully and the fused proteins were expressed in cells.

Objective To explore the surgical treatment of cervical and upper thoracic esophageal carcinoma in patients aged 70 or ol-der. Methods A retrospective analysis were made among 68 patients who were over 70 years old, and all the patients have undergone stom-ach esophagus cervical anastomosis through esophageal carcinoma excision and regional lymph node cleaning. Results All the 68 patients have completed the surgery successfully, and they have recoveried well except 3 cases of death. And the main postoperative complication was cardiopulmonary complication and anastomotic fistula. Conclusion Operative treatment of ercervical and upper thoracic esophageal carcino-ma for elderly patients is characterized with highly difficulty and more complications. Stomach esophagus cervical anastomosis through 2-inci-sion with left chest and left neck could simplify the operative procedure, decrease the postoperative complication and improve the curative effect.

ObjectiveTo investigate the clinical value and pathological basis of peritumoral hyperenhanced rim (PHR) of renal cell carcinomas (RCCs) on CEUS.MethodsCEUS images of 53 patients with 54 renal tumors (27 RCCs,27 renal angiomyolipomas) were analyzed,and the detection and distribution of PHR were evaluated.HE staining and immunohistochemistry of CD34 were performed in tissue surrounding RCCs (TSR) to observe distribution of psuedocapsule,large vessels,and microvasculars among TSR with different modes of PHR.ResultsPHR was found only in RCCs.PHR distribution between RCCs and angiomyolipomas was statistically different (P＜0.05).Using PHR to diagnose RCC,the sensitivity,specificity,positive predictive value,negative predictive value,false positive and false negative was 44.44％ (12/27),100％ (27/27),100％ (12/12),64.29％ (27/42),0 (0/27) and 35.71％ (15/42),respectively.Pseudocapsule distribution between RCCs with PHR and RCCs without PHR was not statistically different (P＞ 0.05).There were rich large blood vessels in TSR with PHR in washin and both phases,and few or thimbleful large vessels were found in TSR without PHR in washout phase.Cancer tissue near the boundary (CTNB) of TSR had the highest microvessel density (MVD).MVD differences in different TSR with PHR were statistically different between washin and washout phases,washin and both phases,both phases with PHR and without PHR (P＜0.05),but no statistical difference was found between washout and both phases (P＞0.05).ConclusionPHR is a highly specific complementary indicator in diagnosing RCC,and it is correlated with rich blood vessels in TSR and (or) a higher MVD value in CTNB.