Objective To establish a method focusing on regulation of CNN3 gene in the rat hippocampus and help to explore the role of CNN3 gene played in the brain physiology and pathology.Methods One cDNA sequence and three shRNAs targeting CNN3 gene were designed and synthesized.The recombinant lentivirus-mediated expressing and three short hairpin RNA ( shRNA) vectors targeting CNN3 gene in the rats were constructed with engineering technology.All recombinant vectors were intravenously injected into rats hippocampi guided by stereotaxic apparatus.Western blot was performed to explore the best shRNA and to study the changes of CNN3 gene in the rat hippocampus after transfection with the silence and over-expressed vectors.Results The lentivirus-mediated vector expressing CNN3-OE and three shRNA vectors targeting CNN3 gene were successfully constructed.Within eight weeks after transfection, the vectors of CNN3-OE and three CNN3-shRNAs changed the expression of CNN3 gene in the rat hippocampus, in particular, all the protein levels of calponin-3 encoded by CNN3 gene were significantly down-regulated along with the time, with the highest inhibitory rate of 73.6%in the CNN3-shRNA2 group.Significant up-regulation of calponin-3 protein level by 93.88%, was found only on the 14th day after transfection.Conclusions Lentivirus-mediated vectors of CNN3-OE and CNN3-shRNAs may regulate in vivo the CNN3 gene level in the local brain region of rats via stereotactic injection.The study lays a foundation for disease prevention and treatment in the future.

Objective To study the distribution of human papillomavirus type 16 (HPV16) variants and their clinical significance in Han women with Cervical Intraepithelial Neoplasia (CIN). Methods Randomly making a collection of DNA samples of cervical cells from 77 Han out-patients infected with HPV16, PCR amplification of HPV16 DNA fragments containing E6 and E7 genes and sequenced. To study the HPV16 variants types in these out-patients and explore the relationship between the HPV16 variants and CIN by comparing the E6 genes sequenced with the reference strains downloaded from the GenBank. Results Among 77 patients, the minimum age is 21 years old, the maximum age is 56 years old, and the average age is 36.39±6.86 years old. 61 patients (accounting for 79.2% ) were diagnosed as CIN Ⅱ and higher grade lesions while 16 patients (accounting for 20.8% ) as CIN Ⅰ. In this research, only European variant and Asian variant were found by Parsimony analyses of the sequences. There are 38 Asian variants and 39 European variants. With χ~2 test, χ~2=0.0034, P=0.9535 > 0.05, it suggested that there was no enough evidence to support Asian- and European- variants had the different risk in the cause of cervical intraepithelial neoplasia and cervical cancer. Conclusion It was not found Asian- and European- variants of HPV16 had different effect on the cervical cancer, but found only two major variants-Asian- and European-variants in Han people in this research. So we have reason to speculate that there are two major HPV16 variants(Asian- and European- variants) in China's Han women, while other variants, especially high cancer-causing Asian/American variant are not common.

OBJECTIVETo evaluate the overexpression of cyclin G1 in cervical intraepithelial neoplasia (CIN) and cervical carcinoma, and the correlation between cyclin G1 and high-risk human papilloma virus (HPV) infection.

METHODSAll of the specimens were obtained from the Department of Pathology of China-Japan Friendship Hospital from January 2000 to August 2004. We detected the expression of cyclin G1 with immunohistochemistry, HPV16/18 infection with in situ hybridization, and high-risk HPV infection with Hybrid capture system II (HC-II) in normal group (25 cases), CIN I (48 cases), CIN II (56 cases), CIN III (54 cases), and invasive cervical squamous-cell carcinoma (SCC, 31 cases).

RESULTSThe positive rates of cyclin G1 expression in CIN (77.85%) and SCC cervical tissues (87.10%) were significantly higher than normal (8.00%, P < 0.01), and the intensities of cyclin G1 expression in CIN (40.60%) and SCC cervical tissues (61.51%) were significantly higher than normal (2.72%, P < 0.05). The positive rates and intensities of cyclin G1 expression increased gradually with the grade of cervical lesions. High-risk HPV infection rates were higher in CIN and SCC than normal groups (P < 0.05). There was a positive correlation between cyclin G1 expression and high-risk HPV infection detected with HC-II (Kendall's tau-b = 0.316, 0.269, 0.352, and 0.474 in CIN I, CINII, CIN III, and SCC, respectively, P < 0.05).

CONCLUSIONSCyclin G1 is overexpressed in CIN and SCC. Cyclin G1 may be a biomarker for detecting CIN and SCC. Cyclin G1 may play an important role in the oncogenesis of CIN and SCC by high-risk HPV infection.

Carcinoma, Squamous Cell ; metabolism ; virology ; Case-Control Studies ; Cervical Intraepithelial Neoplasia ; metabolism ; virology ; Cyclin G ; Cyclin G1 ; Cyclins ; metabolism ; Female ; Human papillomavirus 16 ; genetics ; isolation & purification ; Human papillomavirus 18 ; genetics ; isolation & purification ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Papillomavirus Infections ; metabolism ; virology ; Uterine Cervical Neoplasms ; metabolism ; virology

OBJECTIVETo determine the interaction between 2450-MHz microwaves (MW) radiation and mitomycin C (MMC).

METHODSThe synergistic genotoxic effects of low-intensity 2450-MHz microwave and MMC on human lymphocytes were studied using single cell gel electrophoresis (SCGE) assay (comet assay) and cytokinesis-blocked micronucleus (CBMN) test in vitro. The whole blood cells from a male donor and a female donor were either only exposed to 2450-MHz microwaves (5.0 mW/cm2) for 2 h or only exposed to MMC (0.0125 microgram/mL, 0.025 microgram/mL and 0.1 microgram/mL) for 24 h; and the samples were exposed to MMC for 24 h after exposure to MW for 2 h.

RESULTSIn the comet assay, the comet lengths (29.1 microns and 25.9 microns) of MW were not significantly longer than those (26.3 microns and 24.1 microns) of controls (P > 0.05). The comet lengths (57.4 microns, 68.9 microns, 91.4 microns, 150.6 microns, 71.7 microns, 100.1 microns, 145.1 microns) of 4 MMC groups were significantly longer than those of controls (P < 0.01). The comet lengths (59.1 microns, 92.3 microns, 124.5 microns, 182.7 microns and 57.4 microns, 85.5 microns, 137.5 microns, 178.3 microns) of 4 MW plus MMC groups were significantly longer than those of controls too (P < 0.01). The comet lengths of MW plus MMC groups were significantly longer than those of the corresponding MMC doses (P < 0.05 or P < 0.01) when the doses of MMC were > or = 0.025 microgram/mL. In the CBMN, the micronucleated cell (MNC) rates of MW were 5@1000 and 6@1000, which showed no difference compared with those (4@1000 and 4@1000) of controls (P > 0.05). The MNC rates of 4 MMC groups were 8@1000, 9@1000, 14@1000, 23@1000 and 8@1000, 8@1000, 16@1000, 30@1000 respectively. When the doses of MMC were > or = 0.05 microgram/mL, MNC rates of MMC were higher than those of controls (P < 0.05). MNC rates of 4 MW plus MMC groups were 12@1000, 13@1000, 20@1000, 32@1000 and 8@1000, 9@1000, 23@1000, 40@1000. When the doses of MMC were > or = 0.05 microgram/mL, MNC rates of MW plus MMC groups were much higher than those of controls (P < 0.01). MNC rates of 4 MW plus MMC groups were not significantly higher than those of the corresponding MMC doses.

CONCLUSIONThe low-intensity 2450-MHz microwave radiation can not induce DNA and chromosome damage, but can increase DNA damage effect induced by MMC in comet assay.

Antibiotics, Antineoplastic ; adverse effects ; Cell Culture Techniques ; Chromosome Aberrations ; chemically induced ; Comet Assay ; DNA Damage ; Female ; Humans ; Lymphocytes ; Male ; Micronucleus Tests ; Microwaves ; adverse effects ; Mitomycin ; adverse effects ; Mutagenicity Tests

OBJECTIVETo examine and follow up cervical cytology of pregnant and postpartum women and study their cytopathologic characteristics, so as to determine screening and managing programs for abnormal cervical cytology.

METHODSTotally 5296 patients in pregnancy and postpartum, in which 3729 by computer-assisted cytology test and 1567 by liquid-based monolayers cytology test, were examined and diagnosed by the Bethesda System made in 2001. Those proven epithelial abnormalities patients were followed up until the lesions regressed to normal. The remaining patients, who exhibited persistent abnormalities or progression, were given further examinations.

RESULTSThe positive rate of cervical cytological test was 8.72% (462/5296), including squamous carcinoma (SCA) 1 case (0.02%), high grade squamous intraepithelial lesion (HSIL) 34 cases (0.64%), low grade squamous intraepithelial lesion (LSIL) 145 cases (2.74%), atypical glandular cells (AGC) 5 cases (0.09%), atypical squamous cells cannot exclude HSIL (ASC-H) 14 cases (0.26%), atypical squamous cells of undetermined significance (ASC-US) 263 cases (4.97%). The 419 proven cytological abnormality cases were followed up successfully. The total transnegative rate in three months was 73.74% (309/419), in which 303 cases (72.32%) persisted normal status for more than six months after regression. And the transnegative rate of ASC-US, ASC-H, AGC, LSIL, and HSIL were 79.56%, 64.29%, 100%, 72.14% and 44.12%, respectively. Forty-six cases received biopsy directed by colposcopy. The distribution of coincidence of cytopathologic and histopathologic diagnosis was: SCA 1 case (100%), HSIL 10 cases (76.92%), LSIL 13 cases (65%), ASC-H 2 cases (50%), ASC-US 3 cases (37.50%), total 29 cases (63.04%).

CONCLUSIONSWe should cast more attention to screening cervix lesions in pregnant and postpartum women. Their cytopathologic characteristics are liable to make the clinician give a false positive diagnosis. So we propose to follow up them closely and to lower the indication of biopsy.

Adult ; Biopsy ; Carcinoma, Squamous Cell ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Female ; Follow-Up Studies ; Humans ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; Postpartum Period ; Precancerous Conditions ; pathology ; virology ; Pregnancy ; Uterine Cervical Neoplasms ; pathology ; virology ; Vaginal Smears

OBJECTIVETo detect the high-risk human papillomavirus (HPV) infectious condition in women with abnormal cytology and evaluate its values in the screening of high grade squamous intraepithelial lesion.

METHODSWe used hybrid capture 2 (hc2) method to examine 949 patients with abnormal cervical cytology results [ > or =atypical squamous cells of undetermined significance (ASC-US) according to the 2001 The Bethesda System diagnosis criteria]. All subjects also received colposcopy for tissue studies.

RESULTSAmong 949 patients with abnormal cytology, the diagnoses of atypical squamous cells (ASC), low grade squamous intraepithelial lesion (LSIL), and high grade squamous intraepithelial lesion (HSIL) were made in 432, 310, and 207 patients, respectively. The high-risk HPV positive rate in ASC, LSIL, and HSIL were 40.3%, 44.8%, and 89.4%, respectively. The numbers of patients with pathologically confirmed results of negative intraepithelial lesion or malignancy (NILM), cervical intraepithelial neoplasia 1, 2, 3 (CIN 1, 2, 3), and squamous cell carcinoma (SCC) were 335, 388, 118, 101, and 7, and the high-risk HPV positive rate was 17.3%, 66.2%, 92.4%, 97.0%, and 100%, respectively. Among patients with atypical squamous cells of undetermined significance (ASC-US), rate of HSIL in high-risk HPV positive group and negative group were 10.2% and 0.8%, respectively (P < 0.01). In screening HSIL, the sensitivities of cytology [ > or = ASC cannot exclude HSIL (ASC-H)] and cytology ( > or = ASC-H) plus high-risk HPV testing were 0.925 and 0.991, and the specificities were 0.510 and 0.748, respectively (P < 0.01). Sensitivitives of cytology ( > or = LSIL) and cytology (> or = LSIL) plus high risk HPV in detecting HSIL were 0.898 and 0.982, respectively, while the specificitives were 0. 567 and 0.779, respectively (P < 0.01).

CONCLUSIONSThe positive rate of high-risk HPV increases with the gravity of cervical lesions. In patients with abnormal cervical cytology, high-risk HPV testing can improve the sensitivity and specificity in the screening of HSIL.

Carcinoma, Squamous Cell ; diagnosis ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; diagnosis ; pathology ; virology ; Female ; Humans ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; pathology ; virology ; Risk Assessment ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; virology

Objective To investigate the distribution of HPV 16 variants in Hart women patients without Cervical intraepithelial neoplagia in the diagnosis and treatment center for cervical disease.department of Obstetrics and Gynecology in China-Japan friendship hospital with HPV 16 E5 sequence phylogenetic analysis.Methods PCR amplification of HPV 16 E5 sequences and sequenced.The association between variations types and different cervical lesions was analyzed.Results In this research,We first found that variant classification based on HPV 16 E5 DNA sequence(236 bp)alone had hish rate of accuracy.In addition,for the first time,our research revealed that single-4075T can distinguished the As variant from all other variants.Conclusion If E5 sequence wag used for phylogenetic analysis,it will gready reduce the experimental costs and improve efficiency and cost-effectiveness.For the first time.our research revealed that single 4075T can distinguished the As variant from all other variants.

OBJECTIVETo study the genotoxicity induced by organic bentonite particles in vitro.

METHODSHuman B lymphoblast cells (HMy2.CIR) were exposed to organic bentonite particles at the doses of 0, 1.88, 3.75, 7.50 and 15.00 µg/ml for 24, 48 and 72 h, calcium sulfate (30 µg/ml) and SiO2 (30 and 240 µg/ml) served as negative and positive controls, respectively. The genotoxicity of organic bentonite particles and soluble fraction was detected using comet assay and Cytokinesis-block micronucleus (CBMN) assay.

RESULTSThe results of comet assay indicated that % tail DNA increased with the exposure doses and time in organic bentonite group, % tail DNA at the dose of 15.00 µg/ml for 24 h, 48 h and 72 h in organic bentonite group were 3.20 ± 0.19, 4.63 ± 0.88 and 9.49 ± 1.31 respectively which were significantly higher than those in calcium sulfate group (1.40 ± 0.11, 1.37 ± 0.22 and 0.90 ± 0.16) and those in 30 µg/ml SiO2 group (1.83 ± 0.21, 1.41 ± 0.27 and 2.48 ± 0.25) (P < 0.01). The results of CBMN assay showed that micronucleus frequencies (MNF) in organic bentonite group (except for 1.88 µg/ml for 24 h) were significantly higher than those in 30 µg/ml calcium sulfate group (MNF for 24, 48 and 72 h were 1.33‰ ± 0.58‰, 1.33‰ ± 1.15‰ and 1.33‰ ± 0.58‰) and those in 30 µg/ml SiO2 group (2.00‰ ± 0.00‰, 1.68‰ ± 0.58‰ and 2.33‰ ± 0.58‰) (P < 0.01). The results of two assays demonstrated that the soluble fraction of organic bentonite did not induce the genotoxicity.

CONCLUSIONThe organic bentonite dusts can induce the genotoxicity in vitro, which may be from the particle fraction.

Bentonite ; toxicity ; Cells, Cultured ; Comet Assay ; DNA Damage ; Humans ; Lymphocytes ; drug effects ; Micronucleus Tests ; Mutagenicity Tests ; Quartz ; toxicity

AIM To observe the effects of Sangtongjian Mixture (STJ) on glucose and lipid metabolism,insulin resistance and fat cytokines in type 2 diabetic rats,and their mechanisms of action.METHODS One hundred and forty rats fed on the combination of STZ and high fat diet were established as the type 2 diabetic models.Fasting blood glucose (FBG) level reached more than 16.7 mmol/L and then the rats were randomly divided into model group,metformin (180 mg/kg) group,STJ (73.5,147 and 294 mg/kg) groups.Ten rats were set as the blank group.Each treatment group was intragastrically given the corresponding agents for twelve weeks.The fasting blood glucose levels of rats were measured once every two weeks after the administration.After a 12-week administration period,glycosylated serum protein (GSP),glycosylated hemoglobin (GHb) and lipid profile indices (TC,TG,HDL-C and LDL-C) were determined.The serum insulin level was measured by radioimmunoassay,and homeostasis model assessment of insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) were calculated.The levels of serum adiponectin and leptin were detected by ELISA.RESULTS STJ remarkably decreased the levels of FBG,GSP,GHb,TC,TG,LDL-C,leptin and HOMR-IR in type 2 diabetic rats.Furthermore,STJ also significantly increased the levels of HDL-C,adiponectin and ISI.CONCLUSION STJ can improve glucose and lipid metabolism in type 2 diabetic rats by ameliorating insulin resistance and regulating fat cytokine levels.

Objective To study antioxidant role and mechanism of Rg1 in rats with cerebral ischemia reperfusion injury (IRI).Methods One hundred and twenty healthy male rats were randomly divided into six groups: control group, sham operation group, model group, different concentration (30,60,90 mg/kg) of Rg1 treatment group.MCAO SD rats model was established by suture－occluded method;the Rg1 treatment groups were given Rg1 i.p. in advance, after 24 hours of reperfusion, neurobehavioral scores of all groups were examined by Longa’s standard;The expression of Nrf2 and HO－1 were analyzed by western blot;The content of SOD and MDA was detected by kit.Results The score of model group rats are significantly higher than control group,compared with the model group, the score of different concentration of Rg1 treatment group was decreased (P＜0.05) . The Nrf2 and HO－1 expression in model group was mildly higher than the control or sham group (P＜0.05) . Both of them in every Rg1 treatment group was higher than model group. Compared with control or sham group, SOD content was observably depressed but MDA content was dramatically increased in model group,interestingly,SOD content was enhanced, MDA content was attenuated in different concentration of Rg1 treatment group (P＜0.05). Conclusion Rg1 increases Nrf2 and HO－1 protein expression and SOD content, reduces MDA content,improves neurofunctional performance of rats after MCAO,and alleviates cerebral cerebral IRI.