ObjectiveTo investigate the role and mechanism of microRNA-21 (miR-21) in the proliferation and apoptosis of ovarian epithelial carcinoma cells. MethodsA short-hairpin RNA specifically targeting miR-21 plasmid was constructed, and the recombinant was identified by restriction endonuclease analysis and DNA sequencing. Three experimental groups were included, transfection group (transfected with pSIREN-miR-21 ), negative control group ( transfected with pSIREN-miR-21-neg) and blank control group (without transfection plasmid ). The expression of miR-21 was detected by stem-loop real-time reverse transcription (RT)-PCR in OVCAR3 cells ,and western blot was used to detect the expression of programmed cell death 4 ( PDCD4 ) protein. Tethyl thiazolyl tetrazolium (MTT) and flow cytometry method were used respectively. ResultsRecombinant plasmid (pSIREN-miR-21) was constructed successfully and identified by restriction endonuclease analysis and DNA sequencing. The relative expression level of miR-21 in cells transfection, negative control and blank control group was 0.26 ± 0.08, 1.26 ± 0.21and 1.00 respectively. The level of miR-21 in the cells in transfection group was significantly lower than those in the negtive control and blank control group(P ＜0. 01 ). The gray scale of PDCD4. protein was 1443 ±33,858 ± 19 and 846 ± 16 in the transfection group, negative control and blank control group respectively. The value of PDCD4 in transfection group was higher than other control groups, and there were significantly difference among them( P ＜0. 01 ). Moreover, the optical density of the cells in transfection group was 0. 661 ±0. 015,significantly lower than those in two control groups (0. 848 ± 0. 150 for negative control, 0. 935 ± 0. 133 for blank control,P ＜ 0. 01 ). Forty-eight hours after tranfection, the rate of viable apoptotic cell was significantly higher than negative control and blank control group [(25.821 ± 0. 763 )％ vs.(0. 010 ± 0. 003 ) ％ vs. (0. 238 ± 0. 023) ％ ; P ＜ 0. 01];72 hours after tranfection, the rates of viable apoptotic cell and necrotic cell were all higher than the two control groups [the rate of viable apoptotic cell was ( 30. 480 ±0. 821 ) ％, ( 7. 792 ± 0. 312 ) ％ and ( 7. 033 ± 0. 257 ) ％ respectively ( P ＜ 0. 01 ) ; the rate of necrotic cell was (3.558 ±0.211)％, (1.557 ±0.067)％ and (1.049 ±0.028)％, respectively (P＜0. 01)].ConclusionmiR-21 might play an important role in the proliferation and apoptosis of ovarian epithelial carcinoma cells through negatively control the expression of PDCD4.

Objective To investigate the expression of long intergenic non-protein coding RNA-regulator of reprogramming (Linc-ROR) in high-grade ovarian serous cancer, and explore the relationship between Linc-ROR expression and biological function of high-grade ovarian serous cancer. Methods A total of 34 high-grade ovarian serous cancer tissue samples and 19 normal fallopian tube tissue samples were collected between June 2014 and February 2016. Real-time reverse transcription (RT)-PCR was used to detect the Linc-ROR mRNA expression in different samples. The relationship between Linc-ROR expression level and ovarian cancer International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis was analyzed. Constructed Linc-ROR small interference RNA (siRNA) and pIRES2-EGFP-Linc-ROR plasmid, then Linc-ROR siRNA and pIRES2-EGFP-Linc-ROR plasmid were respectively transfected into SKOV3 cells. Cell proliferation, migration and invasion ability were assessed by cell counting kit-8 (CCK-8), wound healing assay and transwell invasion assay. Results (1) The expression level of Linc-ROR mRNA was significantly higher in high-grade ovarian serous cancer than normal fallopian tube tissues (4.31± 0.38 vs 1.03 ± 0.21; t=25.842, P<0.01). With the progression of FIGO stages, the expression of Linc-ROR was increased (F=95.702, P<0.01), and it was associated with lymph node metastasis (t=7.397, P<0.01). (2) The results of RT-PCR showed that the expression level of linc-ROR in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (0.30 ± 0.11 vs 1.02 ± 0.10; t=15.269, P<0.01). The expression level in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (8.90 ± 0.45 vs 1.03 ± 0.17;t=21.934, P<0.01). The CCK-8 assay showed that when the cells were cultured for 3, 4, 5 and 6 days, the A value in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (P<0.05). And the A value in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (P<0.05). Wound healing assay showed that, after 48 hours incubation, migration rate of cells in Linc-ROR-i group was significantly less than that in the Linc-ROR-NC-i group [(52±4)%vs(67±5)%;t=5.720,P<0.01]. The migration of cells in Linc-ROR-p group was significantly greater than that in the Linc-ROR-NC-p group [(84±4)%vs(66±4)%;t=7.330,P<0.01]. Cell transwell invasion assay showed that, after 48 hours of incubation, the number of invasive cells in Linc-ROR-i group was lower than that in Linc-ROR-NC-i group (74 ± 3 vs 104 ± 3; t=15.810,P<0.01). And the number of invasive cells in Linc-ROR-p group was higher than that in Linc-ROR-NC-p group (217 ± 4 vs 108 ± 5; t=38.060, P<0.01). Conclusion Highly expressed Linc-ROR could enhance the proliferation, migration and invasion ability of high-grade ovarian serous cancer cells, which may be one of the important molecules in the occurrence and development, invasion and metastasis of high-grade ovarian serous cancer.

Objective:To investigate the effects of ginkgo biloba tablets plus relevant mechanism on HPA axis in patients with steroid-dependent asthma.Methods:80 patients with steroid-dependent asthma were divided into control group and experimental group at random,40 cases in every group.The control group were treated with pulmicort respules(1 mg/d) ,patients in the experiment group were given ginkgo biloba tablets in addition to routine treatment(120 mg/d).The therapeutic courses were both 12 weeks.The effective rate of reducing hormone,lung function,the plasma concentrations of ACTH and COR and the levels of PBMC GCR were deter mined before and after treatment.Results:The effective rate of reducing hormone was 89.47%in experimental group,which was 66.67%in control group.There were significant difference at the tows groups(P<0.01).There were no significant difference at lung function,the plasma concentrations of ACTH and COR and the levels of PBMC GCR at pre-treatment( P>0.05).After treatment,the lung function, the levels of plasma concentrations of COR and the levels of PBMC GCR were increased significantly than control group( P<0.01,P<0.05),the levels of plasma concentrations of ACTH was decreased significantly than control group(P<0.01).There was a significant positive correlation between the value of FEV1/FVC and the plasma concentrations of COR,the levels of PBMC GCR( r=0.724,0.632, P<0.01).There was a significant negaitive correlation between the value of FEV1/FVC and the plasma concentrations of ACTH( r=-0.668,P<0.01).Conclusion:Ginkgo biloba tablets combined with routine treatment therapy can increase the effective rate of reducing hormone in patients with steroid-dependent asthma,which can improve the lung function maybe by regulate the HPA axis and upgrade the levels of GCR.

Objective To observe the effect of aerobic exercise preconditioning on growth-associated protein-43 (GAP-43) and Nogo-A in rats after cerebral ischemia/reperfusion. Methods Sprague-Dawley rats were equally divided into sham group (n=40), cerebral ischemia/reperfusion group (n=40) and aerobic exercise preconditioning group (n=40), and global cerebral ischemia model was formed with modified four-vessel occlusion. The rats was sacrificed six hours, one day, three days and seven days after ischemia, respectively. The hippocampus neural cells were observed in five rats with HE staining and immunohistochemistry of GAP-43 and Nogo-A, and the other five rats were test-ed with RT-PCR of GAP-43 and Nogo-A. Results Compared with those in the cerebral ischemia/reperfusion group, the apoptotic neurons and expression of GAP-43 significantly increased all the time points in the aerobic exercise preconditioning group (P<0.01), while the ex-pression of Nogo-A decreased (P<0.01). Conclusion Aerobic exercise preconditioning can promote the regeneration of neuronal cells and axon after cerebral ischemia/reperfusion injury, which is related to the regulation of GAP-43 and Nogo-A.

BACKGROUND: Speech audiometry is decisive in selecting hearing aid,but it is seldom used in clinic in China.OBJECTIVE: To understand the differences in the speech resolution with speech audiometry between in-the-ear (ITE) and behind-the-ear (BTE)hearing aids.DESIGN: Paired controlled study.SETTING: Auditory Center of the 414 Hospital of Chinese PLA, Auditory Rehabilitation Center of Jiangsu Province.PARTICIPANTS: A total of 62 patients with auditory disability were selected from the Auditory Center of the 414 Hospital of Chinese PLA and Auditory Rehabilitation Center of Jiangsu Province. Among 32 ITE hearing aid users, average hearing loss at 500, 1 000, 2 000 and 4 000 Hz, was less than 85 dB, and the patients were aged from 16 to 60 years. Among 30 BTE hearing aid users, average hearing loss at 500, 1 000, 2 000 and 4 000 Hz, was also less than 85 dB, and the patients were aged from 16 to 60 years. All patients were consent.METHODS: Open and close methods were adopted for speech audiometry among ITE and BTE hearing aids users. Monosyllabic and disyllabic words were chosen as the type of speech and 20 words were given to one ear, respectively.MAIN OUTCOME MEASURES: Comparison of speech resolution with monosyllabic and disyllabic words test between ITE and BTE hearing aid.RESULTS: In aspect of the effect of ITE and BTE hearing aids, there were no significant differences of both monosyllabic and disyllabic words no matter through open or close method (P ＞ 0.05). At the opening state, there were significant differences in resolution of monosyllabic and disyllabic words (P ＜ 0.05); while, there were no significant differences (P ＞ 0.05) at the closing state.CONCLUSION: Carrying out speech audiometry in the selection hearing aid is influenced by many factors.

Objective To observe the effect of CO2 pneumoperitoneum combined with position changes on the stability of cardiac electrophysiology in gynecological laparoscopy. Methods The gynecological laparoscopy was performed for 30 patients to undergo elective gynecological laparoscopy under general anesthesia ,with the pneumoperitoneum pressure of 12 mmHg and the Trendelenburg positionat 15° . The observations and analyses were done over the basic monitoring index and the QT interval (QT),T peak tend interval (Tp-e),heart rate corrected QT interval(QTc),QT dispersion(QTd),Tp-e/QT before anesthesia(T0),after anesthesia(T1),1 min after pneumoperitoneum (T2),30 min after pneumoperitoneum and head-down tilt (T3),30 min after deflation and supine position(T4). Results Compared with the time point of T0,QTd increased significantly at T1(P<0.05) and so it was with QT,QTc,QTd,Tp-e,Tp-e/QT at T2,T3,and T4(P<0.05). Compared with the time point of T2,QTc,QTd,Tp-e,Tp-e/QT significantly increased at T3(P < 0.05). Conclusions CO2 pneumoperitoneum combined with Trendelenburg position can prolong ventricular repolarization duration and destroy the stability of cardiac electrophysiology so it may increase the incidence of cardiovascular events.

To explore the changes in metabolites in the greasy tongue coating in patients with chronic gastritis.

Objective To observe the effect of exhaustive exercise preconditioning on GAP?43 and Nogo?A in rats with cerebral ischemia reperfusion. Methods 90 rats were randomly divided into sham oper?ation group,group of cerebral ischemia reperfusion(I/R) and exhaustive exercise preconditioning group.Rats were sacrificed at 6 h,1 d,3 d and 7 d respectively after injury. Neural functions were detected by shuttle box. Morphological changes of hippocampal neural cells were observed by HE staining. Expressions of GAP?43 and Nogo?A were detected respectively by immunohistochemistry and RT?PCR technology. Re?sults Compared with the sham group,the death rate of apoptotic neurons in I/R group was decreased( 6 h:(30.97±2.09)%,1 d:(38.41±1.10)%,3 d:(46.81±2.04)%,1 d:(43.46±1.57)%),the index of learning and memory ability(AARR?7 d:(38.00±12.60)%,PAL?7 d:(27.90±1.79)s) and expression of GAP?43 were decreased(6 h:(2.89±0.85),1 d:(4.06±0.25),3 d:(4.78±0.98),7 d:(7.02±0.21)),the expression of Nogo?A was increased(6 h:(2.93±0.19),1 d:(5.47±0.32),3 d:(4.62±0.26),7 d:(4.12±1.11))(P<0.05).Compared with I/R group,the death rate of apoptotic neurons in exhaustive exercise preconditioning group were decreased,the index of learning and memory ability(AARR?7 d:(20.66±7.60)%,PAL?7 d:(35.53±2.41)s) and expression of GAP?43 were decreased(6 h:(2.03±0.14),1 d:(2.92±0.27),3 d:(3.35±0.34),7 d:(5.24±0.52)),the expression of Nogo?A were increased(6 h:(3.92±0.51),1 d:(6.90± 0.79),3 d:(5.87±0.48),7 d:(5.37±0.50))(P<0.05). Conclusion Exhaustive exercise preconditioning ag? gravates the injury of neurons and neural function,which is related to the regulation of GAP?43 and Nogo?A in the hippocampus of rats.

Objective: To study the relationship of expression of CDX2 and PTEN proteins and gastric metaplasia. Methods: Immunohistochemistry Envision method was used to detect the expression of CDX2 and PTEN proteins in simple intestinal metaplasia (SIM), atypical intestinal metaplasia (AIM) and gastric adenocarcinoma. Results: The positive rates of CDX2 in SIM, AIM and gastric adenocarcinoma was 82.6%, 51.5% and 36.4% respectively. The expression of CDX2 in gastric adenocarcinoma was significantly lower than that in SIM(P0.05). The positive rates of PTEN in SIM, AIM and gastric adenocarcinoma were 86.9%,75.8%and 47.7% respectively and the difference between SIM and gastric adenocarcinoma was significant (P

Objective To elucidate the impact of over-expression of S100A7 on migration,invasion,proliferation, cell cycle, and epithelial-mesenchymal transition (EMT) in human cervical cancer HeLa and CaSki cells.Methods(1)Immunohistochemistry of SP was used to examine the expression of S100A7 in 40 cases of squamous cervical cancer tissues and 20 cases of normal cervical tissues.(2)The vectors of pLVX-IRES-Neo-S100A7 and pLVX-IRES-Neo were used to transfect human cervical cancer HeLa and CaSki cells, and the positive clones were screened and identified. Next, transwell migration assay, cell counting kit-8(CCK-8)assay and fluorescence activating cell sorter(FACS)were used to detect the effect of S100A7-overexpression on the migration, invasion, proliferation and cell cycle of cervical cancer cells. Furthermore, western blot was performed to observe the expression of epithelial marker(E-cadherin)and mesenchymal markers(N-cadherin,vimentin,and fibronectin)of EMT. Results(1)S100A7 expression was significantly higher in cervical squamous cancer tissues(median 91.6)than that in normal cervical tissues(median 52.1;Z=-2.948,P=0.003).(2)Stable S100A7-overexpressed cells were established using lentiviral-mediated gene delivery in HeLa and CaSki cells. S100A7 was detected by real-time quantitative reverse transcription PCR,S100A7 mRNA of S100A7-overexpressed cells were 119 ± 3 and 177 ± 16, increased significantly compared with control groups of median(P<0.01).Compared with the control cells, the number of S100A7-overexpressed HeLa and CaSki cells that passed the transwell membrane assay were increased significanatly(572 ± 51 vs 337 ± 25, P<0.01;100 ± 8 vs 41 ± 4, P<0.01).Matrigel invasion assay showed that the number of S100A7-overexpressed HeLa and CaSki cells that passed the transwell membrane were respectively 441±15 and 110±14,elevated significantly compared with control cells(156±21 and 59±7;P<0.05). However, S100A7 overexpression didn′t influence the proliferation and cell cycle progression of HeLa and CaSki cells(P>0.05). Expression of E-cadherin was dramatically decreased, while N-cadherin, vimentin, and fibronectin increased in S100A7-overexpressed cells. Conclusion S100A7 enhances the migration, invasion and EMT of HeLa cells and CaSki cells, and may be plays an important role in the development of cervical cancer.