OBJECTIVETo compare clinical efficacy differences between"regulating conception-governor vessel" acupuncture method and clomiphene for infertility of polycystic ovarian syndrome (PCOS).

METHODSOne hundred patients of PCOS were randomly assigned into an observation group and a control group, 50 cases in each one. The patients in the observation group were treated with"regulating Conception-Governor Vessel" acupuncture method at Zhongwan (CV 12), Guanyuan (CV 4), Qihai (CV 6), Zhongji (CV 3), Mingmen (GV 4), Yaoyangguan (GV 3) and Yaoshu (GV 2). The acupuncture treatment started at the end of menstruation, once every other day; after four treatments, the follicle was tested with B ultrasound; when follicle was longer than 18 mm or above, the acupuncture treatment was given once a day until follicle was discharged and acupuncture treatment finished. The patients in the control group were treated with oral administration of clomiphene from 5 days into menstruation, 50 mg per day for 5 consecutive days. Both groups were treated for three menstruation cycle. The menstrual cycles, endometrial thickness, endometrium types (A, B and C), cervical mucus scores, basal body temperature (BBT), cases of ovulation, cases of pregnancy were observed.The levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E), prolactin (PRL), progesterone (P) and testosterone (T) in serum were detected before and after the treatment.

RESULTSAfter the treatment, the menstrual cycles were shortened (both<0.05),the endometrial thickness and cervical mucus scores were increased in the two groups (<0.05,<0.01); the improvement of endometrial thickness and cervical mucus score in the observation group were higher than those in the control group (both<0.01). The percentages of type-A endometrium in both groups were higher than those before the treatment (both<0.05); the number of type-A endometrium in the observation group was higher than that in the control group (<0.05). The ovulation rate was 88.0% (44/50) in the observation group, which was superior to 70.0% (35/50) in the control group (<0.05). After treatment, the levels of LH were reduced in the two groups (<0.05,<0.01), which was more significant in the observation group (<0.05).

CONCLUSIONSThe"regulating Conception-Governor Vessel" acupuncture method could improve menstrual cycles, increase endomet-rial thickness and promote the development of follicles in PCOS patient; in addition, it could decrease serum LH level, improve the ovarian functions and increase ovulation rate, which is superior to oral administration of clomiphene.

Objective To investigate the effect of carrimycin on the biological function of pancreatic cancer cells. Methods Pancreatic cancer cell lines MIA PaCa-2, BxPC-3, Panc-1, and PATU 8988 were treated with carrimycin at concentrations of 0 (control group), 2, 4, 8, and 16 μmol/L for 24, 48, and 72 hours. MTT assay was used to measure cell viability; EdU cell proliferation assay was used to observe the effect of carrimycin on DNA replication of pancreatic cancer cells; colony formation assay was used to observe the effect of carrimycin on the proliferation of pancreatic cancer cells; flow cytometry was used to analyze the effect of carrimycin on the cell cycle of pancreatic cancer cells; wound healing assay was used to analyze the effect of carrimycin on the migration of pancreatic cancer cells; Western blot was used to measure the expression levels of the markers such as epithelial-mesenchymal transition (EMT) and cell cycle-dependent protein kinase inhibitor 1A (P21); immunofluorescence assay were used to measure the expression levels of EMT-related markers. An analysis of variance was used for comparison between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the control group, carrimycin significantly inhibited the proliferative activity of MIA PaCa-2, BxPC-3, Panc-1, and PATU 8988 cells in a concentration- and time-dependent manner (all P < 0.01); carrimycin at concentrations of 4, 8, and 16 μmol/L significantly reduced DNA replication in MIA PaCa-2 cells ( t =2.378, 4.984, and 18.970, all P < 0.05) and BxPC-3 cells ( t =4.879, 6.089, and 9.521, all P < 0.01); after treatment with carrimycin at concentrations of 4, 8, and 16 μmol/L, colony formation ability significantly decreased with the increase in drug concentration in MIA PaCa-2 cells ( t =5.889, 11.240, and 15.840, all P < 0.001) and BxPC-3 cells ( t =6.717, 15.800, and 18.850, all P < 0.001). After treatment with carrimycin at concentrations of 4, 8, and 16 μmol/L, there was a significant increase in the proportion of cells in G1 phase in MIA PaCa-2 cells ( t =9.071, 12.280, and 19.360, all P < 0.0001) and BxPC-3 cells ( t =3.061, 4.962, and 8.868, all P < 0.05), and there was a significant reduction in the proportion of cells in S phase in MIA PaCa-2 cells ( t =2.316, 4.165, and 5.562, all P < 0.05) and BxPC-3 cells ( t =2.424, 3.264, and 5.744, all P < 0.05). Western blot further demonstrated that compared with the control group, the expression level of the cell cycle-related protein P21 gradually increased with the increase in the concentration of carrimycin in MIA PaCa-2 cells ( t =5.437, 6.453, and 8.799, all P < 0.001) and BxPC-3 cells ( t =25.130, 44.750, and 52.960, all P < 0.000 1). Wound healing assay showed that after treatment for 12, 24, and 48 hours, carrimycin at concentrations of 0, 4, 8, and 16 μmol/L significantly reduced the lateral migration of MIA PaCa-2 cells (all P < 0.05) and BxPC-3 cells (all P < 0.05). Western blot showed that compared with the control group, carrimycin treatment at concentrations of 4, 8, and 16 μmol/L significantly upregulated the expression of the epithelial marker E-cadherin in MIA PaCa-2 cells ( t =2.388, 4.899, and 5.819, all P < 0.05) and BxPC-3 cells ( t =2.533, 5.836, and 6.774, all P < 0.05) and significantly downregulated the expression of the interstitial marker Snail in MIA PaCa-2 cells ( t =12.440, 14.830, and 16.800, all P < 0.000 1) and BxPC-3 cells ( t=5.039, 5.893, and 7.725, all P < 0.01), and it also significantly downregulated the expression of the interstitial marker Vimentin in MIA PaCa-2 cells ( t =3.105, 7.752, and 11.200, all P < 0.05) and BxPC-3 cells ( t =2.555, 4.883, and 9.153, all P < 0.05). Conclusion Carrimycin can effectively inhibit the proliferation, migration, and EMT process of pancreatic cancer cells, thereby exerting an antitumor biological activity.