FSH receptor binding inhibitor restrains follicular development and possibly attenuates carcinogenesis of ovarian cancer through down-regulating expression levels of FSHR and ERß in normal ovarian tissues.

OBJECTIVES: The current study aimed to investigate FSH receptor binding inhibitor (FRBI) effects in the expressions of FSH receptor (FSHR) and estrogen receptor-beta (ERß) in the mice ovaries at the gene and protein levels, also to find the potential efficacy of FRBI on suppressing ovarian cancer through down-regulating over-expression of FSHR and ERß in the normal ovarian tissues. METHODS: 180 female mice were randomized into six groups (n = 30). Mice of FRBI-1, FRBI-2 and FRBI-3, FRBI-4 were intramuscularly injected with FRBI of 10, 20, 30 and 40 mg/kg, respectively, for five consecutive days. The qPCR and Western blotting were used to determine expression levels of FSHR and ERß mRNAs and proteins in mouse ovaries. RESULTS: The ovarian cortex thickness (OCT) of the FRBI-4 group were less than that FSH group on day 30 (P < 0.05). The numbers of secondary follicles (SF) and the maximum transverse diameters (MTD) of secondary follicles of FRBI-3 and FRBI-4 groups were decreased as compared to FSH group (P < 0.05 or P < 0.01) by 24.11% and 27.47% on day 20 based on the control group (CG) levels. On day 15, the reductions of FSHR mRNA levels in FRBI-2, FRBI-3 and FRBI-4 were 27.78%, 29.37% and 43.65% (P < 0.05 or P < 0.01), respectively in comparison with CG. ERß and FSHR protein levels of FRBI-treated mice were gradually decreased as compared to and CG and FSH group. ERß protein level of FRBI-4 was less than that of CG on day 20 (P < 0.05). On days 15 and 20, estradiol (E2) concentrations of FRBI-2, FRBI-3 and FRBI-4 groups were lower than those of the CG and FSH group (P < 0.05 or P < 0.01). CONCLUSIONS: FRBI could reduce OCT and follicle numbers. A high dose of FRBI (30 mg/kg to 40 mg/kg) could suppress ovarian and follicular development, and attenuate expression levels of ERß and FSHR mRNAs and proteins in the ovaries, additionally inhibit E2 production. Therefore, FRBI will possibly be utilized to restrain the carcinogenesis of ovarian cancer by down-regulating overexpression of FSHR and ERß in the ovaries.

Subunit Vaccine Preparation of Bovine Rotavirus and Its Efficacy in Mice.

BACKGROUND: Rotaviruses (RV) are important viral diarrheal agents in calves. Vaccination is an optimum measure to prevent bovine rotaviruses (BRV) infection. However, little research on BRV VP7 vaccine has been done and currently there is no BRV vaccine. OBJECTIVE: To prepare a subunit vaccine of BRV and investigate its efficacy. METHODS: Total RNA was extracted from MA104 cells infected with bovine rotavirus (BRV) strain GSB01. BRV VP7 gene was amplified using real time fluorescence quantitative PCR (qPCR). The pEASY-T3-VP7 plasmid was digested using HindⅢ and BamHI restriction endonucleases, then recombined into the prokaryotic expression vector pET32a. The pET32a-VP7 and pET32a-VP7-LTB (heat-labile enterotoxin B subunit) were transformed into BL21 (DE3) competent cells of Escherichia coli, respectively, and induced with IPTG, then analyzed using SDS-PAGE. Sixty mice were randomly divided into three groups (n=20). Group A mice was used as His-tag control and mice in group B and C were inoculated with pET32a-VP7 and pET32a-VP7-LTB, respectively. VP7 IgG antibody titers and protection efficiency of pET32a-VP7-LTB were further determined in neonatal mice challenged with GSB01 BRV strain. RESULTS: SDS-PAGE analysis showed that the pET32a-VP7 was highly expressed in the BL21 (DE3) cells. PET32a-VP7 and pET32a-VP7-LTB protein could promote VP7 IgG antibody titer（8.33×103 vs. 17.26×103）in mice. Immunization protection ratios of pET32a-VP7 and pET32a-VP7-LTB proteins in the neonatal mice were 86.4% and 91.7%, respectively. CONCLUSION: The fusion protein of pET32a-VP7-LTB had excellent immunogenicity and protected mice from BRV infection. Our findings can be used for further developing of a high-efficiency subunit vaccine of BRV.

Effect of EDA-A1 gene mutant on proliferation and cell cycle distribution of cultured human umbilical vein endothelial cell ECV304 / 重庆医学

Objective To investigate the effect of ectodysplasin A (EDA‐A1) gene of hypohidrotic ectodermal dysplasia on pro‐liferation and cell cycle of human umbilical vein endothelial cell (ECV304). Methods Recombinant eukaryotic expression vectors pcDNA3. 1(‐)‐EDA‐A1‐M /W (mutant, M and wild, W) containing the coding sequence were transected into ECV304 cells. EDA‐A1 gene was amplified by reverse transcription polymerase chain reaction (RT‐PCR), and the protein was detected by Western blot. Cell viability and cycle distribution were invested by MTT and Flow cytometry (FCM ). Results The EDA‐A1 gene and pro‐tein were detected respectively by RT‐PCR and Western blot in ECV cells transfected with pcDNA3. 1(‐)‐EDA‐A1‐M /W, but not in ECV cells transfected with plasmid pcDNA3. 1(‐) and cells without transection. And also, compared with control groups, EDA‐A1 gene mutant significantly decreased proliferation of ECV cells and its inhibition rate was 45. 70% ( P 0. 05). A significant increase of the G0 /G1 and S fraction was seen in the ECV cells of mutant group, compared with wild group with an accumulation in S phase and a concomitant decrease in G2 /M phase population (P< 0. 05). Conclusion Mutant and wild EDA‐A1 gene may have distinct biological functions on proliferation and cell cycle distribution of cultured human umbilical vein endothelial cell.

Analysis of the infection status of saliva Helicobacter pylori in Lanzhou / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To determine the prevalence of saliva Helicobacter pylori in Lanzhou and investigate Helicobacter pylori-related diseases.</p><p><b>METHODS</b>Helicobacter pylori was detected through bacterial culture, Gram stain microscopy, and urease test from saliva samples collected from 941 residents of Lanzhou. The infection rate and growth of Helicobacter pylori among the residents were analyzed in terms of different oral health conditions, oral disease, gender, urban and rural status, and age.</p><p><b>RESULTS</b>The rate of Helicobacter pylori-positive saliva in Lanzhou was 42.72%. The status of Helicobacter pylori infection showed significant difference among subjects with different oral hygiene and oral diseases. The rate of Helicobacter pylori-positive saliva among females was 47.89%, which was greater compared with the rate among males (38.45%, P = 0.004, chi2 = 8.492). The rate of Helicobacter pylori-positive saliva in the town was 33.99%, which was less than the rate for the villages (50.93%, P = 0.000, chi2 = 27.551). The rate of Helicobacter pylori-positive saliva among residents aged 10 to 59 showed a flat trend with no significant differences. However, the rate of Helicobacter pylori-positive saliva among residents over 60 years old showed a significant increase. No significant difference was found in the growth of saliva Helicobacter pylori (P = 0.086).</p><p><b>CONCLUSION</b>The rate of Helicobacter pylori-positive saliva is related to the subjects' oral hygiene, oral disease, gender, age, and living conditions.</p>

Effects of non-saccharomyces albicans metabolic products on the proliferation of human umbilical vein endothelial cell ECV304 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of non-Saccharomyces albicans metabolic products on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro.</p><p><b>METHODS</b>The parallel dilution supernatant of Saccharomyces tropicalis, Saccharomyces krusei and Saccharomyces glabrata were prepared, and 1, 4, 16-fold(s) diluted concentration and control group were set up. The line of human umbilical vein endothelial cell ECV304 was cultured in vitro and treated by non-Saccharomyces albicans supernatant. The proliferous effect of ECV304 induced by non-Saccharomyces albicans supernatant after 24, 48, 72 h was detected by the methods of MTT, and the changes of cell density and cycle after 48 h were investigated by inverted microscope and flow cytometry.</p><p><b>RESULTS</b>At the 24th hour, all of the higher concentration (1-fold) of non-Saccharomyces albicans supernatant and the 4-folds diluted Saccharomyces krusei could promote ECV304 proliferation(P < 0.05). After adding various non-Saccharomyces albicans supernatant at 48h and 72th hour, Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant significantly increased proliferation rate of ECV304, while Saccharomyces tropicalis supernatant group showed no significant change no matter which concentration was tested. At 48th hour after adding the non-Saccharomyces albicans supernatant, the ECV304 cells density treated by Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant were significantly higher under the inverted microscope. The G0/G1 population of ECV304 cells decreased while cell proliferation index (PI) increased after incubated with Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant for 48 hours (P < 0.05). Saccharomyces tropicalis group showed no significant change (P > 0.05).</p><p><b>CONCLUSION</b>The metabolic products of Sacharoymces krusei and Saccharomyces glabrata could induce proliferation of ECV304 cell, which suggests non-Saccharomyces albicans should be undergone more attention clinically in detection and treatment.</p>

A study on human tongue cancer cells' proliferation affected by Lactobacillus acidophilus / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of Lactobacillus acidophilus (L. acidophilus) on the proliferation and cell cycle distribution of human tongue cancer cells (Tca8113 cells).</p><p><b>METHODS</b>In vitro cultivated human Tca8113 cells were treated by L. acidophilus supernatant, inactivated bacilli, cell free extracts and normal culture medium respectively, which were 1, 4, 16-fold(s) dilutelly, to investigate the proliferous effects of Tca8113 cells using of inverted microscope, cell counting, sulforhodamine B (SRB) and flow cytometry. The free radicals and Ca2+ in Tca8113 cells were also studied by confocal laser scanning microscope (CLSM).</p><p><b>RESULTS</b>At the 48th hour after adding different L. acidophilus components, the Tca8113 cells changed in shape from the diamond-like, polygonal and slabs into the elongated form. In the condition of different times and different culture concentrations, the proliferation of Tca8113 cells was significantly inhibited by L. acidophilus components, which enhanced as the time prolonged and the concentrations of each L. acidophilus components increased according to the cell counting and the SRB experimental analysis. The cell proliferation index (CPI) was significantly reduced (P<0.01). The free radicals and Ca2+ in Tca8113 cells under the effect of each L. acidophilus components for 48 h indicated an obviously rising (P<0.01).</p><p><b>CONCLUSION</b>L. acidophilus restrains the proliferation of Tca8113 cells, which might be due to the increase in quantity of free radicals and Ca2+ in Tca8113 cells, and might be resulted from the release of metabolic products of L. acidophilus.</p>

The correlation between oral colonization of Helicobacter pylori and gastrointestinal disease / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between Helicobacter pylori (H. pylori) colonization in the oral cavity and gastrointestinal disease.</p><p><b>METHODS</b>173 patients with gastrointestinal disease were grouped according to age, gender, periodontal status and types of gastrointestinal disease. H. pylori were detected from saliva samples of all patients by in vitro cultur. The H. pylori-positive rates in different groups were statistically analysed.</p><p><b>RESULTS</b>The H. pylori-positive rate in all patients was 40.46% and the difference between male and female showed significant (P<0.05). The H. pylori-positive rate was 56.72% in the age range 45-64, which was significantly higher than two younger age groups (P<0.05). The H. pylori-positive rate in patients with atrophic gastritis was 77.78%, of which the difference was significantly higher than superficial gastritis group and gastric and duodenal ulcer group respectively (P<0.05). The H. pylori-positive rate in healthy periodontia group was 15.38%, while that in periodontitis group was 72.73% (P<0.05).</p><p><b>CONCLUSION</b>H. pylori is a conditional pathogen. The H. pylori-positive rate from saliva is closely related to the types of gastrointestinal disease in patients, and it is correlated with the periodontal diseases as well. These findings suggest that the oral cavity with periodontal diseases is an ecological niche of H. pylori which might be an important cause for occurrence and re-occurrence of gastrointestinal disease.</p>

In vitro antibiotic effect of the leaching solution of astragalus on cariogenic bacteria / 实用口腔医学杂志

Astragalus produced in Gansu were chosen as the raw material to leachate. Studied the antibiotic effects of the leaching solution on the cariogenic bacteria and compared with the imported bacteriostatic product MI. Streptococcus mutans and Lactobacilli were cultured in the medium for 24 h. The PH and A600 values were measured. Statistical analysis was conducted by using SPSS 13.0. The leaching solution of astragalus has the same inhibitory effects on the growth and acid production of streptococcus mutans and lactobacilli as MI.

An epidemiologic study on candidal transmission from mothers to their neonates in Lanzhou / 实用口腔医学杂志

Objective: To investigate the candidal infection status in puerperas in Lanzhou, and the candidal transmission from mothers to their newborn infants. Methods: Vaginal fluid and saliva samples from 104 puerperas, as well as 104 saliva samples from their newborn infants were collected. The Candida species were cultured, isolated and identified using CHROMagar media. Further identification was done using molecular biological method. Results; In 81 of 312 specimens (104 x2 from mothers and 104 from infants), Candida species were found. 39.42% (41 cases) was observed in the vaginal fluid and 33.65% (35 cases) was in saliva of puerperas respectively, and 21. 15% (22 cases) in both vagina and oral cavity. 4.81% (5 cases) was found in oral cavities of newborn infants. The distribution of Candida species were 53 Candida albicans, 33 Candida glabrata, 2 Candida krusei and 1 Candida tropical. In 2 pairs of mother-infant, the same genotype of Candida ablicans was identified using PCR method. Conclusion; The Candida detection rate of newborn infants and transmission rate from mothers to their neonates in Lanzhou are higher than that reported in other areas. The colonization of Candida in newborn infants is relevant to both horizontal and vertical transmission. It can decrease the possibility of Candidal infection in newborn infants by controlling the Candidal transmission in hospital and preventing the infection in pregnant women.