Extracellular vesicles of Bifidobacterium longum reverse the acquired carboplatin resistance in ovarian cancer cells via p53 phosphorylation on Ser15.

We previously found that the relative abundance of Bifidobacterium was increased after chemotherapy; however, the role of Bifidobacterium longum in chemotherapeutic drug resistance in ovarian cancer (OVC) remains unclear. This study aimed to understand the potential effects and mechanism of B. longum extracellular vesicles (B. longum-EVs) on carboplatin (CBP) resistance in OVC. Eight normal and 11 ovarian tissues were collected and the expression of B. longum genomic DNA and its association with acquired CBP resistance in OVC patients was determined. After isolating EVs by ultracentrifugation from B. longum (ATCC 15707), CBP-resistant A2780 cells were treated with PBS, CBP, B. longum-EVs, or CBP + B. longum-EVs, and subsequently analyzed by CCK-8, Edu staining, Annexin V/PI double staining, wound healing, and Transwell assays to detect cell viability, proliferation, apoptosis, migration, and invasion, respectively. MRP1, ATP7A, ATP7B, and p53 expression as well as p53 phosphorylation were measured by western blot analysis. S15A mutation of p53 was assessed to examine the potential role of p53 Ser15 phosphorylation in CBP-resistant OVC. B. longum levels were elevated and positively associated with CBP resistance in OVC patients. Only high concentrations of B. longum-EVs attenuated A2780 cell proliferation, apoptosis, migration, and invasion. B. longum-EVs exposure significantly enhanced the sensitivity of CBP-resistant A2780 cells to CBP and decreased the expression of drug resistance-related proteins. The effect of B. longum-EVs on reversing CBP resistance was completely inhibited by S15A mutation of p53. B. longum-EVs enhanced the sensitivity of OVC cells to CBP through p53 phosphorylation on Ser15.

[Clinical Effect of Bushen Quyu Recipe Combined with Acupuncture in Treatment of Clomiphene- resistant Polycystic Ovary Syndrome Infertility Patients after Cold Needle Puncture Drainage Operation].

Objective To observe the clinical effects of Bushen Quyu Recipe (BQR) combined with acupuncture in treatment of clomiphene-resistant polycystic ovary syndrome (PCOS) infertility pa- tients after cold needle puncture drainage operation. Methods Totally 170 clomiphene-resistant PCOS in- fertility patients were recruited from March 2011 to October 2013, who were assigned to the control group and the observation group according to random blocking method, 85 cases in each group. Patients in the control group received cold needle puncture drainage operation alone, while those in the observation group additionally took BQR and received acupuncture after cold needle puncture drainage operation. Clinical efficacy was observed in the two groups before and after treatment. The spontaneous ovulation rate and the pregnancy rate were followed-up. The levels of serum sex hormones and hermodynamic indicators of ovarian blood flow were detected in the two groups before and after treatment. Results Successful pregnancy occurred in 63 cases of the observation group, significantly better than that of the con- trol group (52 cases; x² =7. 63, P <0. 05). The spontaneous ovulation rate was 75. 29% at month 3 of follow-ups and 88. 24% at month 6 of follow-ups in the observation group, significantly higher than those of the control group [56. 47% , 67. 06%; x² =6. 70, X² =10. 98, P <0. 05). In the observation group the total pregnancy rate was 74. 12% , higher than that of the control group [61. 18% ; X² =4. 46, P <0. 05). Compared with before treatment in the same group, levels of luteinizing hormone (LH) , testesterone (T) , estradiol (E2) significantly decreased in the two groups after treatment; levels of follicular stimulating hormone (FSH) , peak systolic velocity (PSV) , end diastolic volume (EDV) obviously increased in the two groups after treatment (P <0. 05). The decrement of T, LH, E2 levels and the increment of FSH, PSV, EDV levels were obviously higher in the observation group than in the control group (P <0. 05). Conclusion BQR combined with acupuncture in treatment of clomiphene-resistant PCOS infertility pa- tients after cold needle puncture drainage operation could effectively promote the recovery of menstruation, elevate the success rate of pregnancy, and was helpful to improving levels of sex hormones and ovarian blood perfusion.

[Value of high risk human papilloma virus detection in screening and diagnosing cervical lesion in littoral of Zhejiang province].

OBJECTIVE: To analyse the infection of high-risk human papiliomavirus (HR-HPV) in cervical lesion wome, and evaluate the significance of high-risk human pappilomavirus detection by hybrid capture II (HV-II) in screening and diagnosing cervical lesion, especially high grade cervical intraepithelial neoplasia (CIN). METHODS: A series of 1130 patients of cervical lesion were preliminarily diagnosed by cervical cytological examination, HR-HPV detection by HC-II , colposcopy and biopsy under the colposcopy between June 2009 and December 2008, including 212 CIN I and (or) condyloma (CIN I/HPV I), 442 CIN II/III, 28 invasive cervical cancer. cervical cytological examination is by thin prep liquid-based cytology test(TCT),and HR-HPV detection is by HC-II. RESULTS: In 1130 cases the positive of HR-HPV was 65.84% (744/1130). Unusual cytology result were 862 cases, with 356 ASCUS, 84 ASCH, 216 LSIL, 184HSIL and 22 cancer. The number of biopsy > or = CINI/HPVI was 682, positive rate of HR-HPV was 78.59% (536/682). In screening CIN II or above, sensitivity, specificity, PPV and NPV of TCT were 88.94%, 32.73%, 48.49%, 80.60%, of HR-HPV DNA detectiort by HC-II were 90.21%, 51.82%, 57.14%, 88.14%, and of HR-HPV detection combined with cytology were 97.45%, 22.42%, 47.22%, 92.50%. CONCLUSION: The infection rate of HR-HPV in cervical lesions is higher in each age group. Infection rate of HR-HPV is ascending with serious degree of cervical lesion. HR-HPV detection by HC- II is an important method in screening cervical lesion. HR-HPV detection is a viable option in the management of women with ASCUS and LSIL of TCT, with higher sensitivity and NPV.

[TTV and HPV co-infection in cervical smears of patients with cervical lesions in littoral of Zhejiang province].

OBJECTIVE: To investigate the prevalence of transfusion-transmitted virus (TTV) and human papillomavirus (HPV) co-infection in cervical smears of patients with cervical lesions in littoral of Zhejiang province and analysis of transmitted route. METHODS: Nested polymerase chain reaction (nPCR) was established. TTV DNA were tested by nPCR in cervical smears of 95 patients with cervical lesions and 55 healthy women, paired serum samples were available from 55 and 42 women, and their viral titer. The genotypes of 95 specimens of cervical cytology were detected with HybriMax. The phylogenetic group of TTV was determined by means of nPCR with N22 primers. RESULTS: The prevalence of TTV DNA in cervical smears of patients with cervical lesions and healthy women was 52.7% (29/55) and was comparable with that in paired serum sample (50%). Symptomatic women had significantly higher prevalence of TTV DNA in cervical smears (74.7%) than healthy controls (P = 0.005). The TTV DNA prevalence in patient serum samples was 51%. The phylogenetic groups of TTV serum isolates were concordant with those of TTV from cervical smears of the same subjects, and genotype was G1b. The TTV viral titer in cervical smears were 10 to 1000 times as high as in serum. The total infection rate of HPV was 98.9% in patients, and was 27.3% in healthy women. The frequently detected genotype was HPV16, 18, 33 of HSIL, and HPV6 of LSIL. The HPV positive study subjects had significantly higher TTV DNA prevalence than HPV negatives (P = 0.02). CONCLUSION: High prevalence of TTV in cervical smears suggests that sexual transmission is another mode of expansion of TTV infection among the population. The higher viral titer in cervical smears than in the respective serum samples might indicate active TTV replication in the female genital tract. Nevertheless, cooperation between TTV and HPV needs to be further investigated.