Small hairpin RNA targeting inhibition of NF-κB gene in endometriosis therapy of Macaca fascicularis / 中华妇产科杂志

Objective To observe the therapeutic effect of NF-κB gene short hairpin RNA (shRNA) on endometriosis and identify the function of NF-κB on the maintenance and development of endometriosis in Macaca fascicularis.Methods The Macaca fascicularis model of endometriosis was developed,which divided into experimental group,negative control group and simple model group.The high specificity adenovirus vector mediated shRNA targeting NF-κB gene and negative control shRNA adenovirus with no-load NF-κB gene were synthesised.The experimental group injected the adenovirus which carried the NF-κB shRNA into the endometriosis lesions under laparoscopy surgery,the negative control group with no-load shRNA adenovirus and the simple models group injected with normal saline.Four weeks later after the injection,an observed operation was performed through laparoscopy and some lesions were collected.The CD34 immunohistochemistry of these lesions were done to detect the microvessel density,then the variation of the microvessel density among each group were observed.The expression of the NF-κB and proliferating cell nuclear antigen (PCNA) were detected through western blot.Results First,the Macaca fascicularis model of endometriosis was successful developed,and the experimental group has an evident atrophy in ectopic lesions compared with the previous.The lesions' microvessel density in experimental group decreased evidently compared with the negative control group and simple model group (0.002 0±0.000 3 versus 0.021 9±0.002 6 versus 0.024 5±0.003 3),and the differences was statistically significant (P＜0.01).The expression of PCNA (0.37±0.17 versus 0.57±0.26 versus 0.57±0.28) and NF-κB (0.338 ± 0.174 versus 0.678 ± 0.021 versus 0.645 ±0.098) in experiment group was lower than the negative control group and simple model group,the differences were statistically significant (all P＜0.01).Conclusion Through targeting suppressed the NF-κB gene expression by NF-κB shRNA,we can inhibit the development of endometriosis through reducing the ability of angiogenesis and cell proliferation of ectopic endometrial cells.

Adenovirus vector-mediated short hairpin RNA targeting nuclear factor-κB suppresses proliferation of endometrial cells of Macaca fascicularis in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the effect of a high specific adenovirus vector-mediated shRNA targeting nuclear factor-κB (NF-κB) on cell proliferation of the endometrium of Macaca fascicularis.</p><p><b>METHODS</b>The adenoviral vector NF-κB-p65-shRNA and the empty vector were separately trasnfected in cultured endometrial cells of Macaca fascicularis. The changes in the expression of the target gene protein and apoptotic proteins, cell proliferation, and cell cycle distribution were observed after the transfection.</p><p><b>RESULTS</b>Compared with the control cells, infection of the endometrial cells with the NF-κB-p65-shRNA adenovirus significantly increased the expression levels of apoptotic proteins, promoted apoptosis of the endometrial cells, and reduced the cells in division?stage.</p><p><b>CONCLUSIONS</b>NF-κB-p65 shRNA adenovirus can effectively promote apoptosis of endometrial cells and inhibit the proliferation of endometrial cells of Macaca fascicularis.</p>

Effect of interleukin-1β on expressions of activin A and its related factors in cultured endometrial stromal cells from patients with endometriosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of interleukin-1β (IL-1β) on the expressions activin A, follistatin, and cripto in cultured human endometrial stromal cells (HESCs) form patients with endometriosis.</p><p><b>METHODS</b>Cultured HESCs were stimulated with 250, 500, and 750pg/ml IL-1β, and the mRNA and protein expressions of activin A, follistatin, and cripto were assayed using real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>IL-1β treatment caused significant dose-dependent increments of the mRNA and protein expressions of activin A and follistatin and of the mRNA expression of cripto in cultured HESCs.</p><p><b>CONCLUSION</b>IL-1β can affect the expressions of activin A, follistatin and cripto in HESCs from patients with endometriosis.</p>

Apoptosis of human primary ovarian granulose cells infected with lentivirus carrying bcl-2 gene / 中国组织工程研究

BACKGROUND: Lentivirus can infect divided and undivided cells. It remains uncertain whether the lentivirus can successful y infect primary ovarian granulosa cells. OBJECTIVE: To investigate infecting ratio and cel apoptosis of lentivirus carrying bcl-2 gene in primary human ovarian granulose cells cultured in vitro. METHODS: The lentiviral vector carrying bcl-2 gene was constructed using molecular biology, and packaged into lentivirus with high titer. The resulting recombinant lentivirus carrying bcl-2 genes were then used to infect primary human ovarian granulosa cells in vitro at different multiplicity of infection, 10, 50, 100, 200, and 400. Infection efficiency and cel proliferation were observed at 24, 48, 72, and 96 hours fol owing infection. Cel apoptosis was detected by flow cytometry, and bcl-2 gene transcription was assessed using reverse transcription PCR. RESULTS AND CONCLUSION: Primary human ovarian granulosa cells adhered at 24 hours, and exhibited polygon- or fusiform-shape and colony-like growth. When multiplicity of infection was 100, cel appearance and growth remained unchanged, and infection efficiency was high, which reached the peak up to 72 hours. Moreover, the positive rate was up to 60% in granulosa cells. Lentivirus carrying bcl-2 gene could increase expression of Bcl-2 protein and inhibit apoptosis of primary ovarian granulosa cells.

Illumina sequencing 16S rRNA tagging reveals diverse vaginal microbiomes associated with bacterial vaginosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze and compare vaginal microbiomes in healthy women at child-bearing ages and patients with bacterial vaginosis (BV).</p><p><b>METHODS</b>A total of 74 vaginal swabs of the vaginal fornix were collected from 37 BV patients and 37 healthy women. BV status was assessed according to Amsels clinical criteria for all the subjects and confirmed using Gram-stain criteria (Nugent scores). Genomic DNA of the samples was extracted for amplifying the 16S rRNA V6 hypervariable region by PCR and pyrosequencing by Illumina. BIPES, UCHIME, TSC and GAST were employed to analyze the information of the species from the samples.</p><p><b>RESULTS</b>Lactobacillus was the predominant species in healthy women (more than 95%), including mainly L. iners and L. crispatus, with a small quantity of Gardnerella, Granulicatella, Streptococcus, Prevotella, Escherichia and other genus. The α diversity was significantly increased in 30 BV patients (P<0.001), and β diversity also changed obviously shown by decreased Lactobacillus (varying from 45% to 1%, consisting mainly of L. iners) or even absence Lactobacillus in 6 cases, with increased relative abundance of Gardnerella, Prevotella, Granulicatella, Anaerococcus, Parvimonas, Peptoniphilus.harei, Peptostreptococcus, and Dialister. Different from previous data, 7 BV cases showed a predominance of the rare species L.gasseri and L.acidophilus (75% to 50%).</p><p><b>CONCLUSION</b>Lactobacillus is the predominant vaginal species in healthy women (mainly L. iners and L. crispatus) co-existing with many other bacteria and a variety of microorganisms. Lactobacillus is significantly decreased and even absent in most of BV patients, and some cases show the predominance of the rare species L.gasseri and L.acidophilus.</p>

Effect of folic acid in preventing aberrant methylation of fetal endometriosis susceptibility gene HOXA10 / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect aberrant methylation in the promoter region of fetal endometriosis susceptibility gene homeobox-10 (HOXA10) in women with and without folic acid supplementation and explore the effect of folic acid in optimizing intrauterine environment.</p><p><b>METHODS</b>Thirty-six cord blood specimens were collected between January, 2010 and December, 2012 from pregnant women with endometriosis, including 22 with folic acid treatment and 15 without. Methylation-specific polymerase chain reaction (MSP) and bisulfite salt modified sequencing (BSP) were employed to detect aberrant methylation of HOXA10 gene in these specimens.</p><p><b>RESULTS</b>The methylation rate of HOXA10 gene differed significantly between pregnant women with endometriosis taking folic acid and those who did (P<0.05).</p><p><b>CONCLUSION</b>Folic acid treatment can significantly reduce the methylation rate of fetal endometriosis susceptibility gene HOXA10.</p>

Effect of cervical conization through hysteroscopy for cervical intraepithelial neoplasia III / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of cervical conization through hysteroscopy in the treatment of cervical intraepithelial neoplasia (CIN) III.</p><p><b>METHODS</b>Seventy-four patients with CIN III underwent cervical conization through hysteroscopy (TCRC group), and 65 received cold knife conization (CKC group). The operating time, volume of blood loss, concordance rate with pathology, recurrence rate, rate of cervix adhesion and pregnancy rate were compared between the two groups.</p><p><b>RESULTS</b>The operating time, mean blood loss, cure rate, and recurrence rate were 15.1∓3.2 min, 12.5∓1.8 ml, 94.6%, and 5.4% in TCRC group, respectively, as compared with those of 25.8∓3.8 min, 21.6∓2.4 ml, 81.5%, and 18.5% in CKC group, all showing significant differences between the two groups (P<0.05).</p><p><b>CONCLUSION</b>Compared with CKC, TCRC has such advantages as less blood loss, shorter operating time, more accurate lesion localization, fewer complications, higher cure rate, and lower recurrence rate without significant adverse effect on pregnancy.</p>

The clinical significence of determination of serum vascular endothelial growth factor and endostatin in endometriosis / 医学研究生学报

Objective:To investigate the wether vascular endothelial growth factor (VEGF) and endostatin could be used as new serum markers of endometriosis. Methods:Preoperative serum levels of VEGF and endostatin in 82 women with endometriosis(stage Ⅰ-Ⅱ 43 cases and stage Ⅲ-Ⅳ 39 cases) were assayed in duplicate using enzyme linked immunoadsordent assay. Serum levels were compared with levels of 60 healthy controls. Results: Serum VEGF and endostatin levels in endometriosis were significantly higher than that of controls. The levels of VEGF and endostatin of endonmetriosis stages Ⅰ-Ⅱ were significantly lower than those of stage Ⅲ-Ⅳ. The sensitivity, specificity, positive predictive value and negative predictive value of serum VEGF in detecting endometriosis were 0.92,0.78,0.82 and 0.90, respectively; those of enedostatin were 0.95, 0.84, 0.86 and 0.95, respectively. When both factors were used concomitantly, they were 0.98,0.75,0.76 and 0.98, respectively. Conclusion:These results indicates that the balance of angiogenic stimulators and inhibitors may regulate the development and progression of endometriosis and demonstrates that the circulating levels of both VEGF and endostatin maybe useful markers for endometriosis.

Comparison between curettage and hysteroscopic electroresection for endometrial polyps / 中国微创外科杂志

Objective To compare the effects between dilation and curettage (D&C) and hysteroscopic electroresection (HE) in the management of endometrial polyps. Methods Entered into the study there were 86 patients with endometrial polyps, 32 of them were treated by D&C (Group D&C) and 54 by HE (Group HE). The operation time, pre- and post- operative complications, and recurrence of 2 groups were compared. Results Of the Group D&C and Group HE, the operation time were (8.5?4.2) min and (9.0?3.1) min, without statistically significant differences (t=0.632, P=0.529), while the recurrence numbers and recurrence time of the 2 groups were 9 (36.0%) and 3 (7.5%) cases, (5.3?3.5) months and (11.2?4.8) months, respectively, with significant differences (?2=6.516, P= 0.011; t=6.058, P=0.000). Conclusions Compared with D&C, HE is characterized by lower recurrence rate and longer recurrence time.