Effect of combined treatment with acupuncture, moxibustion and medication on endometrial receptivity and expression of serum HOXA10 in polycystic ovary syndrome of kidney deficiency and blood stagnation / 中国针灸

OBJECTIVE@#To compare the clinical effect of the combined treatment of acupuncture, moxibustion, Chinese herbal medicine and western medication and simple western medication on polycystic ovary syndrome (PCOS) of kidney deficiency and blood stagnation pattern and explore the effect on endometrial receptivity and the expression of serum homeobox gene A10 (HOXA10).@*METHODS@#A total of 60 patients with PCOS of kidney deficiency and blood stagnation pattern were randomized into a combined treatment group and a western medication group, 30 cases in each one. In the western medication group, on the fifth day of menstruation, clomiphene citrate tablets were taken orally, 50 mg each time, once daily, consecutively for 5 days. On the day when the follicle diameter was ≥ 18 mm, chorionic gonadotrophin for muscular injection, a dose of 10 000 U was given. Before sleep, the aspirin enteric-coated tablets were taken orally, 50 mg (except during menstruation). In the combined treatment group, on the base of the treatment as the western medication group, acupuncture and moxibustion were adopted and the Chinese herbal for tonifying the kidney and activating blood circulation was taken orally. The acupoints were Guanyuan (CV 4), Qihai (CV 6), Zusanli (ST 36), Sanyinjiao (SP 6), Zigong (EX-CA 1), etc. Acupuncture was remained for 30 min each time, once every two days and discontinued during menstruation. Chinese herbal was given from the 3rd day of menstruation till the onset of the next menstruation, one dose each day. After consecutive treatment for 3 menstrual cycles in the two groups, the real-time polymerase chain reaction (RT-PCR) method was adopted to determine the expression of serum HOXA10 before and after treatment in the patients of the two groups. The endometrial thickness at ovulatory phase, uterine arterial flow 7 days after ovulation [including uterine arterial pulsatility index (PI), resistance index (RI), peak systolic velocity (PSV)/end diastolic velocity (EDV), meaning S/D], pregnancy rate and the score of Chinese medicine symptoms before and after treatment were compared in the patients between the two groups.@*RESULTS@#① After treatment, the expression of serum HOXA10 was higher than that before treatment in the patients of the two groups (@*CONCLUSION@#The combined treatment with acupuncture, moxibustion and medication effectively improves endometrial receptivity and uterine arterial flow in the patients with PCOS of kidney deficiency and blood stagnation pattern and increases pregnancy rate. The therapeutic effect is better than the simple western medication and its mechanism is probably related to the regulation of serum HOXA10 expression.

Expression of HOXA10 gene and its biological effects in endometrial carcinoma / 中国肿瘤生物治疗杂志

@# Objective: To study the expression of H O X A 1 0 gene in endometrial carcinoma and its effect on the apoptosis, migration and invasion of Ishikawa cells. Methods: Twenty-one cases of endometrial carcinoma tissue samples and 25 cases of normal endometrial tissue samples from patients treated at the Department of Obstetrics and Gynecology, Nanjing Drum Tower Hospital from 2012 to 2013 were collected for this study. The mRNA and protein expressions of H O X A 1 0 in endometrial carcinoma and normal endometrial tissues were separately tested by Realtime-qPCR (qRT-PCR) and Western blotting. Ishikawa cells were infected with adenovirus-flagHOXA10 at different multiplicity (5, 10, 20 MOI), and infected by adenovirus-flag-lacz (20 MOI) as control; And the cell apoptosis was tested by Flow Cytometry. Ishikawa cells were transfected with 50 nmol/L si-HOXA10 plasmids and 50 nmol/L si-NC plasmids, as down-regulation group and down-regulation control group, respectively. Ishikawa cells were infected with 20 MOI adenovirus-flagHOXA10 and 20 MOI adenovirus-flag-lacz, as up-regulation group and up-regulation control group, respectively. The ability of migration and invasion was detected by transwell assay. Results: The results of qRT-PCR and Western blotting showed that the expressions of H O X A 1 0 mRNA and protein in endometrial carcinoma samples were both significantly lower than normal samples [mRNA: (0.56± 0.14)vs (1.36±0.33), P<0.01; protein: (1.01±0.25) vs (2.10±0.71), P<0.001]. After the up-regulation of H O X A 1 0 gene in Ishikawa cell line, the cell apoptosis rate in ad-flag-HOXA10 groups (5, 10, 20 MOI) was significantly raised, and most of which was in the early apoptosis [(50.92±8.79)%, (55.17±4.07)%, (76.10±3.65)% vs (7.74 ± 0.15)%, all P <0.01]. The number of migrated cells was markedly up-regulated in si-HOXA10 group [(248±25) vs (135±15), P <0.01] but markedly down-regulated in ad-flag-HOXA10 group [(50±6) vs (100±13), P <0.01]. The number of invasive cells was markedly up-regulated in si-HOXA10 group [(131±18) vs (66±9), P <0.01] but markedly down-regulated in ad-flag-HOXA10 group [(34±8) vs (60±4), P <0.01]. Conclusions: Both mRNAand protein expressions of H O X A 1 0 were down-regulated in endometrial carcinoma samples than in normal endometrium. Up-regulation of H O X A 1 0 gene in Ishi kawa cell line can promote cell apoptosis and inhibit cell migration and invasion.

Sequential therapy combined with ovulation induction of PCOS in endometrium of infertile women / 中成药

AIM To observe the endometrium receptivity of polycystic ovary syndrome patients and research advance of expression of HOXA10 in endometrium.METHODS Eighty PCOS patients were divided into treatment group and control group,40 persons in each group randomly.The control group was treated with clomiphene (CC) + human menopausal gonadotropin (HMG) + human chorionic gonadotropin (HCG),and the treatment group was treated with CC + HMG + HCG + Yin-Nourishing Yang-Supplementing.After 3 periods,take a record for below:endometrial thickness (Em) in the middle,advanced stage and mid-secretory phase of hyperplasia endometrii,the levels of spiral artery PI/RI in midluteum endometrium,the levels in serum of E2 and P in midluteum endometrium,the expression of HOXA10 mRNA from both groups,clinical pregnancy rate and abortion rate for each group.RESULTS Compared with the control group,endometrial thickness in the treatment group was increased and there is statistical difference in the middle and late follicle phases (P ＜ 0.05),but there was no statistical difference in midluteum endometrium (P ＞ 0.05);the level of E2 and P in the treatment group were raised (P ＜0.05) with statistical significance;PI and RI were obviously contracted (P ＜ 0.01).Expression of HOXA10 mRNA was increased (P ＜ 0.01).Compared with the control group,the treatment group had a significant difference in pregnancy rate (P ＜ 0.05).The abortion rate was lower,but there was no statistical significance (P ＞ 0.05).CONCLUSION Yin-Nourishing Yang-Supplementing Treatment can obtain higher pregnancy rate and lower abortion rate of PCOS patients and the mechanism might be associated with raising the expression of HOXA10 mRNA and reducing spiral artery PI/RI,also improving the function of corpus luteum treatment,then to improve the receptivity of endometrium.Yin-Nourishing Yang-Supplementing Treatment is well worth popularizing in further clinical application.

Regulatory Effects of Zishen Yutai Pills on Expression Levels of HOXA10 and Its Downstream Gene EMX2 in Endometria of Ovulation-inducing Mice at Different Implantation Stages / 广州中医药大学学报

Objective To investigate the regulatory effects of Zishen Yutai Pills on the expression levels of homeboxA10 (HOXA10) and its downstream target gene empty spiracles homebox 2 (EMX2) in the endometria of ovulation-inducing mice at different implantation stages. Methods Seventy-five estrous female Kunming mice were randomly divided into 5 groups, namely normal group, model group 1, model group 2, treatment group 1, treatment group 2, 15 mice in each group. The model group 1 was given short-term protocol for ovulation induction; the model group 2 was given long-term protocol for ovulation induction; the treatment group 1 was given Zishen Yutai pills (at the dose of 0.4 g/mL) on the basis of the protocol for the model group 1; the treatment group 2 was given Zishen Yutai Pills (at the dose of 0.4 g/mL) on the basis of the protocol for the model group 2; the normal group was given intragastric administration or intraperitoneal injection of the same volume of normal saline. The mRNA and protein expression levels of HOXA10 and EMX2 in mouse uterus were detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot method, respectively. Results Compared with the normal group, the mRNA and protein expression levels of HOXA10 were decreased, and the mRNA and protein expression levels of EMX2 were increased in model group 1 and model group 2(P< 0.01). Compared with the corresponding model group 1 and 2, the mRNA and protein expression levels of HOXA10 were significantly up-regulated (P < 0.01) , and the mRNA and protein expression levels of EMX2 were decreased in the treatment group 1 and 2 (P < 0.01), respectively. Conclusion Zishen Yutai Pills may improve mouse endometrial receptivity by up-regulating HOXA10 expression and inhibiting EMX2 expression.

Expression and mechanism of action of miR-196a in epithelial ovarian cancer

Objective To explore the expression, biological function and possible mechanism of action of microRNA molecular-196a (miR-196a) in epithelial ovarian cancer. Methods RT-PCR was used to detect the expression quantities of epithelial ovarian tissue, benign ovarian tissue, normal ovary epithelial tissue, ovarian cancer cell lines and miR-196a in normal ovarian epithelial cells to analyze the relationship between the expression of miR-196a and the clinical pathologic parameters of ovarian cancer. Among those cell lines, the cell line of which miR-196a expressed the most or least was selected and transfected the ovarian cancer cell line by using negative control plasma and miR-196a inhibitor. After transfection, RT-PCR was used to test the expression quantity of miR-196a, Transwell chamber method was applied to determine the migration and invasion abilities of ovarian carcinoma cells and Western blot was employed to detect the expression of HOXA10 protein. Results The relative expression quantities of miR-196a in ovarian cancer tissue and benign ovarian tissue were significantly higher than that in normal ovarian epithelial tissue, and the expression quantity of miR-196a in ovarian cancer tissue was distinctively higher than that in benign ovarian tissue (P 0.05). Compared with normal ovarian epithelial cell line IOSE80, the expression quantities of miR-196a of all ovarian cancer cell lines increased obviously and differences were statistically significant (P < 0.05). Among them, the expression of miR-196a of ovarian cancer cell line SKOV3 was the highest, while it decreased significantly (4.678 ± 0.785 vs. 2.131 ± 0.345, t = 2.938, P < 0.05) after the ovarian cancer cell line SKOV3 was transfected by miR-196a inhibitor. The results of Transwell chamber method showed that the migration and invasion abilities of ovarian cancer cells SKOV3 were declined significantly after the expression of miR-196a was down-regulated and the difference showed statistical significance (P < 0.05). The results of Western blot revealed that the relative expression of HOXA10 decreased distinctly after the expression of miR-196a was down-regulated and also the difference showed statistical significance (P < 0.05). Conclusions The miR-196a might serve as a cancer-promoting gene to promote the migration and invasion of epithelial ovarian cancer by downstream target gene HOXA10.

Expression and mechanism of action of miR-196a in epithelial ovarian cancer

OBJECTIVE@#To explore the expression, biological function and possible mechanism of action of microRNA molecular-196a (miR-196a) in epithelial ovarian cancer.@*METHODS@#RT-PCR was used to detect the expression quantities of epithelial ovarian tissue, benign ovarian tissue, normal ovary epithelial tissue, ovarian cancer cell lines and miR-196a in normal ovarian epithelial cells to analyze the relationship between the expression of miR-196a and the clinical pathologic parameters of ovarian cancer. Among those cell lines, the cell line of which miR-196a expressed the most or least was selected and transfected the ovarian cancer cell line by using negative control plasma and miR-196a inhibitor. After transfection, RT-PCR was used to test the expression quantity of miR-196a, Transwell chamber method was applied to determine the migration and invasion abilities of ovarian carcinoma cells and Western blot was employed to detect the expression of HOXA10 protein.@*RESULTS@#The relative expression quantities of miR-196a in ovarian cancer tissue and benign ovarian tissue were significantly higher than that in normal ovarian epithelial tissue, and the expression quantity of miR-196a in ovarian cancer tissue was distinctively higher than that in benign ovarian tissue (P  0.05). Compared with normal ovarian epithelial cell line IOSE80, the expression quantities of miR-196a of all ovarian cancer cell lines increased obviously and differences were statistically significant (P < 0.05). Among them, the expression of miR-196a of ovarian cancer cell line SKOV3 was the highest, while it decreased significantly (4.678 ± 0.785 vs. 2.131 ± 0.345, t = 2.938, P < 0.05) after the ovarian cancer cell line SKOV3 was transfected by miR-196a inhibitor. The results of Transwell chamber method showed that the migration and invasion abilities of ovarian cancer cells SKOV3 were declined significantly after the expression of miR-196a was down-regulated and the difference showed statistical significance (P < 0.05). The results of Western blot revealed that the relative expression of HOXA10 decreased distinctly after the expression of miR-196a was down-regulated and also the difference showed statistical significance (P < 0.05).@*CONCLUSIONS@#The miR-196a might serve as a cancer-promoting gene to promote the migration and invasion of epithelial ovarian cancer by downstream target gene HOXA10.

Influence of targeted silence HOXA10 gene on leukemia NB4 cells / 白血病·淋巴瘤

Objective To explore the effects of lentivirus-mediated RNA interference targeting HOXA10 gene on the proliferation, apoptosis and drug resistance of leukemic cell line NB4.Methods NB4 cells were divided into three groups: interference group, negative control group and untreated group.The infection efficiency of lentivirus for NB4 cells was detected by flow cytometry, and the expression of HOXA10 gene of NB4 cells at mRNA and protein level was detected by real-time PCR and Western blot.Cell survival was determined by MTF assay, and apoptosis and necrosis rates were detected by flow cytometry.Western blot was used to detect the influence of down-regulation HOXA10 gene on the multi-drug resistance-1 (MDR-1) protein.Results The ratio of GFP positive cells was up to 90 ％.HOXA10 gene mRNA and protein levels were decreased in interference group compared with control group.The inhibition rate of interference group was (52.12±4.02) ％, the apoptosis rate of interference group was (30.0±2.7) ％, and their differences in the interference group and in control groups (negative control group and untreated group) were significant (P ＜ 0.05).Western blot results showed that interfering HOXA10 gene significantly reduced the resistance gene MDR-1 expression level and reverse the drug-resistant of leukemia cells.Conclusions Lentivirns-shHOXA10 can steadily reduce the expression level of HOXA10, inhibit the leukemic cells proliferation, promote apoptosis and reverse drug-resistant.HOXA10 gene is expected to become a new target for reversing leukemia drug resistance.

Regulatory Effect of Estrogen, Progestin and HB-EGF on the Expression of HOXA10 Gene in Ishikawa Cells / 华中科技大学学报（医学）（英德文版）

HOXA10 gene plays an essential role in differentiation of the endometrium and in human reproduction. The aim of this study was to investigate the regulatory effect of sex steroids and HB-EGF on HOXA10 gene in Ishikawa cells. Ishikawa cells were incubated with 17-beta estradiol (10-8 mol/L), medroxyprogesterone acetate (MPA) (10-6 mol/L), RU486 (10-5 mol/L) or HB-EGF (10 ng/mL) for 48 h respectively. The expression of HOXA10 gene was detected by immunofluorescence,reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. Our results showed that either estrogen alone, progestin alone or progestin combined with estrogen could significantly increase the expression of HOXA10 gene 48 h after the treatment (P＜0.05). But estrogen combined with progestin and RU486 could inhibit the up-regulation by estrogen and progestin. HB-EGF could elevate the expression of HOXA10 gene 48 h after the treatment (P＜0.05). It is concluded that both estrogen and progestin can up-regulate the expression of HOXA10 gene in Ishikawa cells, but RU486 can inhibit the effect and HB-EGF can elevate the expression level of HOXA10 gene.

Expression of HOXa-10 gene in endometrium mice during early pregnancy / 重庆医科大学学报

Objective:To investigate the effect of HOXa-10 gene in the mouse uterus during embryo implantation. Method: real-time fluorescence quantitative PCR (FQ-PCR) and immunohistochemistry were used to detect the HOXa-10 gene and protein of the mouse of un-pregnant pregnant d1,d2,d3,d4,d5 and d7. Result:The HOXa-10 mRNA/?-actin mRNA expression in pregnant group is higher than that of non-pregnant grnup, and the results showed gradual hoist trend as days past, the HOXa-10 mRNA expression reached the maximum at pregnant d4. Immunohistochemical analysis showed the same results as FQ-PCR. Conclusion:HOXa-10 is persistently expressed in mouse endometrium during the early pregnancy and maybe participates in the regulation process of mouse blastodyst implantation.

THE FUNCTION OF HOXa-10 GENE IN THE COURSE OF MOUSE BLASTOCYST IMPLANTATION / 解剖学报

Objective To investigate the function of HOXa-10 gene in the course of mouse blastocyst implantation. Methods Real-time fluorescence quantitative PCR was applied to detect the expression of HOXa-10 gene in normal mice and day1,day2,day3 day4,day5 and day7 pregnant mice.Plasmid containing the HOXa-10 cDNA fragment with liposome and plasmid containing HOXa-10 antisense oligonucleotides with liposome were transfected respectively into the uterine horns of day 2 pregnant mice,and the number of blastocyst-implanted was counted and compared with that of normal day 2 pregnant mouse. Results HOXa-10 gene expression in the pregnant group is higher than that of the non-pregnant group,and the results showed a gradual increasing trend as days went by: HOXa-10 gene expression was the highest on pregnancy d4,began descending on pregnancy d5 and was close to that of non-pregnancy on pregnancy d7.The number of blastocyst increased in the uterine horns which were injected with plasmid containing the HOXa-10 cDNA fragment with liposome.On the contrary,the number of blastocyst decreased in the uterine horns which were injected with plasmid containing HOXa-10 antisense oligonucleotides with liposome(P