OBJECTIVE: To explore the inhibitory effects of silencing long non-coding RNA (LncRNA) HIF1A-AS2 on epithelialmesenchymal transition (EMT) and tumor stem cell-like phenotype in cervical cancer cells. METHODS: We designed 3 shRNA constructs for silencing HIF1A-AS2 in CaSki cells, and the shRNA with the strongest interference effect was selected for subsequent experiment. CaSki cells were transfected with shRNA-NC or Sh-HIF1A-AS2, and the changes in cell viability, invasion ability, EMT, expressions of EMT-related proteins, formation of cell spheres and expressions of stem cell markers were detected. RESULTS: Transfection with shRNA-NC and Sh-HIF1A-AS2 did not significantly affected the viability of CaSki cells ( CONCLUSIONS Silencing HIF1A-AS2 can inhibit proliferation, invasion and migration of cervical cancer cells
Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Epithelial-Mesenchymal Transition/genetics* ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; RNA, Long Noncoding/genetics* ; RNA, Small Interfering/genetics* ; Uterine Cervical Neoplasms/genetics*

To compare the clinical effect of needling the Ashi point (4cm lateral to the lower border of the spinous process of cervical vertebra) plus Yaotongdian on the hand and point injection on the Ashi point with Triamoninde. The curative effect of acupuncture was better than that of point injection.

Objective To study the influence of alprostadil on the serum adiponectin and leptin in patients with early diabetic nephropathy.Methods 98 patients with early diabetic nephropathy were randomly divided into two groups,the control group(n =48 cases) and the treatment group(n =50 cases).The patients of the control group were treated through the conventional treatment.However,the patients of the treatment group were treated plus alprostadil.They were treated for 21 days.The urinary albumin excretion rate (UAER),adiponectin,leptin and insulin resistance were detected before and after treatment.Results The UAER,adiponectin leptin and insulin resistance were improved after treatment,while those of the treatment group were more significantly improved than those of the control group(P ＜ 0.05 or P ＜ 0.01).Conclusion Alprostadil can improve the conditions of early diabetic nephropathy which is through improving the adiponectin and insulin resistance.

Background and Objectives: This study was to investigate the role of microRNA-29a-3p (miR-29a-3p) in human bone marrow mesenchymal stem cells (hBMSCs), and its relationship with steroid-associated osteonecrosis. Methods: and Results: The online tool GEO2R was used to screen out the differentially expressed genes (DEGs) in GSE123568 dataset. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-29a-3p, forkhead box O3 (FOXO3), alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (OCN) and RUNX family transcription factor 2 (Runx2) in the hBMSCs isolated from the patients with steroid-associated osteonecrosis. CCK-8 assay was executed to measure cell viability; western blot assay was utilized to detect FOXO3, ALP, Runx2, OCN and β-catenin expression. Cell apoptosis and cell cycle were detected by flow cytometry. Immunofluorescence assay was used to detect the sub-cellular localization of β-catenin. Bioinformatics analysis and luciferase reporter gene assay were performed to confirm whether miR-29a-3p can combine with FOXO3 3’UTR. MiR-29a-3p was markedly up-regulated in the hBMSCs of patients with steroid-associated osteonecrosis, while FOXO3 mRNA was significantly down-regulated. Transfection of miR-29a-3p mimics significantly inhibited the hBMSCs’ proliferation, osteogenic differentiation markers’ expressions, including ALP, Runx2, OCN, and repressed the ALP activity, as well as promoted cell apoptosis and cell-cycle arrest. FOXO3 was identified as a target gene of miR-29a-3p, and miR-29a-3p can inhibit the expression of FOXO3 and β-catenin, and inhibition of miR-29a-3p promoted translocation of β-catenin to the nucleus. Conclusions MiR-29a-3p can modulate FOXO3 expression and Wnt/β-catenin signaling to inhibit viability and osteogenic differentiation of hBMSCs, thereby promoting the development of steroid-associated osteonecrosis.

Objective To explore the clinical significance and the dynamic changes of serum anti-inflam-mation factor interleukin-10(IL-10)and inflammatory factor interleukin-8(IL-8)in cerebral infarction.Methods The serum levels of IL-10 and IL-8 in 30 patients were measured by double-antibody sandwich enzyme-linked inunu-nosorbent assay(ELISA)on days 1.3,7 and 14 respectively after infarction.Results The serum levels of IL-10 and IL-8 were higher in patients than those in control group on days 1,3 and 7(P<0.05).Conclusion IL-10 and IL-8 are involved in the pathophysiology of acute cerebral infarction.

Objective To study the effects of serum C-type natriuretic peptide (CNP) ,insulin-like growth factor II (IGF-Ⅱ ) , endothelin (ET) ,neuron-specific enolase (NSE) and S100B protein(S100B) on the prognosis of the patients with traumatic brain injury .Methods A total of 110 patients with craniocerebral trauma admitted in our hospital from January 2016 to January 2017 se-lected as the craniocerebral trauma group and further divided into the mild ,moderate and severe craniocerebral trauma groups ac-cording to the Glasgow Coma Scale (GCS) .Then the levels of serum CNP ,IGF-Ⅱ ,ET ,NSE and S100B in all cases were analyzed by enzyme-linked immunosorbent assay (ELISA) .Their influence on the prognosis of the patients with craniocerebral trauma and the correlation among various indicators were analyzed .Results The levels of CNP and IGF-Ⅱat admission in the craniocerebral trauma group were significantly decreased ,while the levels of ET ,NSE and S100B were significantly increased ,the difference com-pared with the control group was statistically significant (P<0 .05) .Serum CNP and IGF-Ⅱlevels in the death group ,plant survival group and disabled group were significantly decreased .The difference was statistically significant (P<0 .01) .Serum CNP and IGF-Ⅱlevels in the moderate and severe craniocerebral trauma groups were gradually increased with the disease course progress ,while serum ET ,NSE and S100B levels were gradually decreased with the disease course progress ,the difference was statistically signifi-cant(P<0 .05) .In the patients with craniocerebral trauma ,the positive correlation existed between CNP and IGF-Ⅱ ,between ET and S100B ,between ET and NSE ,and between NSE and S100B(P<0 .01) ,while the negative correlation existed between IGF-Ⅱand ET ,between IGF-Ⅱ and S100B ,between CNP and ET ,and between IGF-Ⅱand NSE (P<0 .01) .Conclusion Serum CNP , IGF-Ⅱ ,ET ,NSE and S100B are correlated to the severity of craniocerebral trauma ,which has a higher clinical application value for judging the disease condition ,evaluating the prognosis in cradiocerebral trauma .

Objective:To investigate the application value of sound touch quantification (STQ) in screening high risk group of non-alcoholic steatohepatitis(NASH).Methods:From February 2019 to January 2020, 98 patients with non-alcoholic fatty liver disease(NAFLD) and 36 healthy volunteers were included in this study. NAFLD patients were divided into NASH high risk group and NASH low risk group. The liver Young′s modulus were measured by STQ and compared among the three groups. The ROC curve was used to analyze the effectiveness of STQ in screening NASH high risk population. When the maximum value of Jordan index was selected, the best boundary threshold was selected to analyze its sensitivity, specificity and accuracy. Liver biopsy was performed in 15 patients with abnormal liver function as validation group.Results:There were significant differences in the mean values of Young′s modulus among high risk group, low risk group and control group ( F=33.068, P<0.01). The Young′s modulus of NASH high risk group was significantly higher than those in NASH low risk group and control group ( P<0.05). The Young′s modulus between NASH Low risk group and control group was not significantly different( P=0.443). The best screening threshold was 6.46 kPa, the sensitivity was 89.2%, the specificity was 85.2%, and the accuracy was 88.6%. In the verification group, when the Young′s modulus≥6.46 kPa, the accuracy was 73.3%, the sensitivity was 75.0%, and the specificity was 66.7%. Conclusions:As a screening index, STQ elastic imaging technology has certain value in screening NASH high risk group.

ObjectiveTo observe the therapeutic effect of Yiyuan Qiwei pills （YYQW） on diabetes mellitus-induced induced erectile dysfunction （DMED） in rats and explore its regulation on the nitric oxide （NO）-cyclic guanosine monophosphate （cGMP） signaling pathway. MethodFifty-five healthy SD male rats of clean grade aged 2-3 months underwent intraperitoneal injection of streptozotocin （STZ） to induce the DMED model， and another 10 healthy SD male rats of clean grade aged 2-3 months were assigned to the control group. The model rats were randomly divided into a model group， a sildenafil group （5 mg·kg^-1， ig）， and low-， medium-， and high-dose YYQW groups （1.5， 3.0， 6.0 g·kg^-1， ig）. The rats in the model group and the control group were given normal saline by gavage at 10 mL·kg^-1， once a day for two months. After intervention， the penile erectile function of rats in each group was measured by a pressure detection system. The pathological changes and ultrastructure of penile corpus cavernosum were observed by hematoxylin-eosin （HE） staining and transmission electron microscopy， respectively. The level of NO in the corpus cavernosum was detected by nitrate reductase. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of cGMP and advanced glycation end products （AGEs）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of endothelial nitric oxide synthase （eNOS）， neurogenic nitric oxide synthase （nNOS）， total nitric oxide synthase （NOS）， and phosphodiesterase type5 （PDE5） in rat penile tissues. The expression of above proteins was detected by Western blot. ResultCompared with the control group， the model group showed decreased intracavernous pressure （ICP）， NO， and cGMP levels， reduced mRNA and protein expression of nNOS and NOS， and increased PDE5 mRNA and protein expression （P<0.05）. Compared with the model group， the sildenafil group and the YYQW groups displayed increased ICP， NO， and cGMP levels， elevated mRNA and protein expression levels of nNOS and NOS， and reduced PDE5 mRNA and protein expression levels （P<0.05）. There were no pathological changes in the tissues and cell ultrastructure of the corpus cavernosum in the control group， while serious pathological changes were observed in the model group. Additionally， the sildenafil group and the YYQW groups were superior to the model group， the optimal effect was observed in the high-dose YYQW group. ConclusionYYQW can improve the penile erectile function of DMED rats and reduce the pathological damage of corpus cavernosum. The mechanism may be related to the promotion of nNOS and NOS expression， the inhibition of PDE5 expression， and the activation of the NO/cGMP signaling pathway.

OBJECTIVE To establish and apply a method for simultaneous determination of the contents of dicentrine， protopine and coptisine in Tibetan Corydalis pallida of different origins， and to provide reference for origin determination and quality control of the kind of medicinal materials. METHODS Ultra performance liquid chromatography-tandem quadrupole mass spectrometry method was used. The determination was performed on Agilent EC-C18 column （100 mm×2.1 mm， 2.7 μm） with mobile phase consisted of acetonitrile-0.1% formic acid by gradient elution. The flow rate was 0.2 mL/min， and the column temperature was set at 35 ℃ . MS detection was carried out by electrospray ionization in positive modes， multiple reaction monitoring mode was used for quantitative analysis. RESULTS The injection mass concentrations of dicentrine， protopine， coptisine ranged from 5.88 to 117.60， 53.70 to 1 074.00， and 4.85 to 97.00 ng/mL， respectively， showing a good linear relationship with their respective peak areas （r＝0.998 2， 0.991 9， and 0.999 6， respectively）. The limits of quantitation were 2.35， 1.07 and 1.46 ng/mL； the limits of detection were 1.17， 0.54， 0.49 ng/mL， respectively. RSDs of precision， stability （24 h） and repeatability tests were all lower than 2.0％. The average recovery rates were 97.41%， 98.89% and 105.44%（ all RSDs＜5.0％， n＝6）. CONCLUSIONS The established method has good selectivity and high accuracy， and is suitable for the rapid analysis of dicentrine， protopine and coptisine in Corydalis. The total contents of three alkaloids in different original medicinal materials are from high to low in order of C. chrysosphaera， C. mucronifera， C. pygmaea， C. hendersonii and C. conspersa. The alkaloid contents in C. chrysosphaera and C. mucronifera are relatively similar， but no dicentrine has been detected in C. conspersa and C. hendersonii.