Objective: To sum up characteristics of hotline counseling about mental health problems in Shanghai. Method: All records of hotline counseling from 1990 to 2000 were input into computer. Retrospective analysis was done. Results: In the past 10 years, the main issues in hotline counseling were associated with love affairs (18.1%), emotional troubles (15.8%), psychosis (11.3%), and interpersonal relationship (8.1%), which were also related to the help-seekers background, such as gender, age, education levels, occupation and marriage status.

Objective:To screen the plasma microRNAs( miRNAs) of differential expression in a high fat diet-induced insulin resistance in mouse models;further investigations on the mice with insulin resistance treated by TLR4 inhibitors TAK-242,and to study the changes of plasma miRNAs expression profile and the relationship among TLR4, miRNAs and high fat diet-induced insulin resistance. Methods:The plasma samples were from 3 mouse groups of previous study,namely,the control group with general basic diet ( low fat diet,LFD) ,TLR4 inhibitors TAK-242 treatment group with a high fat diet ( HFD-T) and the high fat diet control group( HFD-C) . The differential expressed miRNAs was screened by expression profiling of plasma miRNAs, which was detected using mouse miRNA microarray. The quantitative Real-Time PCR ( qRT-PCR ) was used to verify the results of microarray. The target genes of differential expressed miRNAs were predicted in TLR4 signaling pathway using bioinformatics methods,and the GO and KEGG database molecular annotation system were used to investigate the main effects of the miRNAs targeted genes on the biological functions or signal pathway. Results:The screening results of miRNA microarray chip showed that,comparing miRNAs expression between HFD group and LFD control group,185 miRNAs were significant in the high fat diet group,including 6 up-regulated and 179 down-regulated miRANs. A significant difference of miRANs was also found between HFD-T group and LFD control group,the total number of differential expression miRNAs was 171,and all of them were down-regulated. Comparing miRNAs expression between HFD-C group and HFD-T group,13
miRNAs were significant in HFD-T group,all of them were down-regulated. Bioinformatics analysis results showed that a total of 10 in-teraction proteins with TLR4 were predicted;the difference of mmu-miR-3095-3p,mmu-miR-5113,mmu-miR-709 and mmu-miR-335-3p expression levels was more than 1 000 times between HFD-C group and HFD-T group,and their target genes can be found in TLR4-in-teraction protein or Toll like receptor signaling pathway;GO and KEGG analysis showed 74% of these target genes belonged to the biological processes genes, and the transcription factors accounted for 82%. The expression of mmu-miR-3095-3p, mmu-miR-5113, mmu-miR-709 and mmu-miR-335-3p detected by qRT-PCR exhibited the similar patterns of down regulation to those shown in microarray results. Conclusion:When insulin resistance occurs,there is a change in plasma miRNAs expression profile,this change is associated with TLR4 and its signaling pathways. The finding enrichs the possible mechanisms of insulin resistance and provides a basis for finding miRNAs diagnostic markers for early diagnosis of insulin resistance.

Objective To develop a new Two-step magnetic resonance electrical impedance tomography(MREIT)algorithm based on radial basic function(RBF)neural network for imaging electrical impedance distribution of a head.Methods Firstly,the magnetic resonance imaging(MRI)system with high resolution was used to set up 3D model of the object and to identify the boundaries of different tissues.Then RBF MREIT algorithm was applied to estimate piece-wise homogeneous impedance values of those tissues,respectively.Furthermore,the impedance of each element within each region of the FEM model was estimated according to the RBF genetic algorithm method based on the piece-wise constant impedance.Results Computer simulations were conducted in a three-sphere head model(scalp-skull-brain,SSB)and the simulation results showed the applicability and feasibility of the present Two-step MREIT algorithm in imaging continuous electrical impedance distribution within the head.Conclusion The present Two-step MREIT algorithm is an effective method for imaging the continuous electrical impedance distribution within the human head.

Objective To evaluate the impact of cerebrospinal fluid drain(CSFD) on the incidence of acute spinal cord injury(SCI) following extensive TAAA repair.Methods From February 2009 to July 2016,153 patients underwent extensive TAAA repairs with a consistent strategy of normal thermia,non-circulatory bypass,sequential aortic cross clamping,aortic-lilac bypass,and intercostal artery reconstruction.The repairs were performed with preoperative CSFD (n =78) or without CSFD (n =75).In the former group,CSFD was inserted after the patient has been anaesthetized and continued for 72 hours after surgery.The target CSF pressure was 10 mmHg or less.Results The mean age of patients was (38 ± 10) years and 108 (70.6％) were male.There were 87 (53.8％) patients with previous aortic surgeries and 33 (22％) with Marfan syndrome.The two groups had similar risk factors for paraplegia.Aortic clamp time,operation time and number of reattached intercostal arteries were similar in both groups.In-hospital mortality rates were 1.3％ (one patient) and 6.7％ (five patients) for CSFD and the group without CSFD,respectively (P =0.086).Ten patients (13.3 ％) in the group without CSFD had paraplegia develop.In contrast,only two patients in the CSFD group(2.6％) had postoperative paraplegia(P =0.013).Stepwise logistic regression analysis identified CSFD had spinal cord protection,P =0.026;OR =0.171;95％ CI:0.036-0.809).No patients occurred CSF catheter related complications.Conclusion This randomized clinical trial showed that preoperative CSFD placement could be an effective strategy in preventing SCI following extensive aortic aneurysm repair.Care should be taken to prevent complications related to overdrainage.

OBJECTlVE To estabIish a simpIe,sensitive and quick method for determination of B7011 in rat pIasma. METHODS The method of protein precipitation with methanoI was used for pre-treatment of pIasma sampIes determined by Iiquid chromatography mass spectrometer. The Iinear reIa-tionship,intra-batch and inter-batch precision,specificity,matrix effect,recovery rate,the accuracy and stabiIity of the pIasma sampIes were vaIidated. The concentration of B7011 in pIasma was determined by LC-mS/ mS foIIowing a singIe intravenous injection of B7011 0.5 mg·kg-1 to rats. RESULTS The Iinear range of B7011 was 30-20 000 μg·L-1 ,the Iower Iimit of quantification was 30 μg·L-1 in pIasma,the in-tra-batch precision of 60,1000,16 000 and 10 000 ng·mL-1 was 5.61% -13.31%,2.31% -8.35%, 2.02%-9.47% and 4.0%-15.0% respectiveIy,and inter-batch precision was 10.05%,2.55%,3.75% and 8.58% respectiveIy. The recovery of 60,1000,and 16 000 μg·L-1 was 114.12%,109.2% and 101.06%respectiveIy. The average peak concentrations were 8373.28 and 8564.59 μg·L-1 ,the mean AUC was 98 400 and 104 000 μg·L-1·h and the t1/ 2z for B7011 was 41.7 and 63.6 h in bIood of maIe and femaIe rats, respectiveIy. CONCLUSlON The estabIished method is sensitive, fast and simpIe and concentration of B7011 in pIasma is determined by LC-mS/ mS foIIowing a singIe intravenous injection of B7011 0.5 mg·kg-1 to rats. It can satisfy the requirements of pharmacokinetic and toxicokinetic studies.

Objective To observe the effect of nitric oxide (NO) on exflagellation of malaria parasite. Methods The level of parasitemia and gametocytemia in DBA/2 mice infected with Plasmodium yoelii 17XL was measured by scanning Giemsa-stained blood smears, and the NO level in culture supernatant of splenocytes was checked using Griess reaction. The mice were injected with different doses of NO donor (NOC5) on day 4 post-infection, and control mice were injected with NOC5 precursor. On day 6 post-infection, mice were injected with NOS inhibitor (L-NMMA), and control mice were injected with D-NMMA and PBS, respectively. Blood samples were collected from tail vein of mice before injection, 30 and 60 min after being injected with NOC5 and NOC5 precursor, 4 and 8 h after being injected with L-NMMA, D-NMMA, and PBS respectively. Exflagellation number of gametocytes in blood culture was counted under microscope. Results The NO level in culture supernatant of splenocytes from mice on day 4 and 6 post-infection was 16.5 mmol/L and 30.4 mmol/L, and exflagellation number was 11.33 and 0.66, respectively. The number of exflagellation in parasitized erythrocytes, obtained from mice on day 4 post-infection, was 5.33 and 2.66, respectively, 30 and 60 min after injection of 1 mg NO donor (NOC5), significantly lower than that of the control (P

Objective To explore the significance of long non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) in hepatocellular carcinoma (HCC),to predict the target gene of TUG1,and to provide a ref-erence for further study of TUG1 in HCC.Methods The differential expression of TUG1 in HCC was ana-lyzed by using the UALCAN database and the survival analysis of TUG1 was performed.The target gene of TUG1 was predicted by RegRNA 2.0 biology software,HMDD,targetscan and microT-CDS,and the regulato-ry network of lncRNA TUG1-microRNAs-mRNAs was constructed.The predicted target gene was analyzed by Gene Ontology (GO) and KEGG signal transduction pathway enrichment by using FunRich platform. Results TUG1 expression in HCC was significantly increased,and the expression level of TUG1 increased generally with the increase of tumor grade.The overall survival of patients with low expression of lncRNA TUG1 was significantly longer than that of lncRNA TUG1 high expression patients.There were four possible binding sites of HCC related microRNAs (hsa-mir-122-5p,hsa-mir-200a-3p,hsa-mir-34c-3p,hsa-mir-629-3p) on TUG1,which regulated 245 downstream target genes and formed the regulatory network of lncRNA TUG1-microRNAs-mRNAs.In the biological process,microRNA target genes were highly enriched in the processes such as the regulation of nucleobase,nucleoside,nucleotide and nucleic acid metabolism.In KEGG pathway analysis,microRNA target genes were highly enriched to the signal pathways mediated by Syndecan and TRAIL.Conclusion TUG1 expression level in HCC increased.Increased expression of TUG1 is associat-ed with poor prognosis in HCC.Bioinformatics methods can be used to explore the mechanism of tumorigene-sis from the molecular level,which can provide valuable information for subsequent experiments and clinical diagnosis and treatment.

Objective The aims of the study were to investigate the effects of human islet amyloid polypeptide (hIAPP) on autophagy in INS-1 cells and its underlying mechanism,and to explore the role of autophagy in hIAPP-induced cytotoxicity and oxidative stress.Methods INS-1 cells were treated with hIAPP (10 μmol/L) for 24 h in the presence or absence of N-acetyl-L-cysteine (NAC),compound C,5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) and 3-methyladenine (3-MA),respectively.Transmission electron microscopy was used to observe the number of autophagosome in cells.Cell viability was determined by methyl thiazolyl tetrazolium (MTT) test.2',7'-dichlorofluorescin diacetate (DCFH-DA) assay was used to measure the relative levels of reactive oxygen species (ROS).Western blot was used to detect expression of adenosine monophosphate-activated protein kinase (AMPK) and autophagic markers p62 and microtubule associated protein 1 light chain3 (LC3).Results Treatment of INS-1 cells with hIAPP resulted in a significant increase in the number of autophagosomes and the expression of LC3-Ⅱ/LC3-Ⅰ (both P＜0.05).Meanwhile,treatment of INS-1 cells with hIAPP enhanced the level of ROS to 1.76 times of control cells (P＜0.01).Co-treatment with NAC,an antioxidant,inhibited hIAPP-induced ROS generation,and the expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK in the INS-1 cells (all P＜0.05).Pretreatment of INS-1 cells with AMPK inhibitor compound C suppressed hIAPP and AICAR,an activator of AMPK,induced expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK (all P＜0.05).Autophagic inhibitor 3-MA and compound C aggravated the hIAPP-induced cell death and ROS generation in INS-1 cells (All P＜0.05).The cytotoxic effects of hIAPP were significantly attenuated by co-treatment with AICAR (P＜0.05).Conclusion Autophagy may act as an adaptive mechanism to alleviate hIAPP-induced oxidative damage and toxicity in INS-1 cells.

Objective To construct the luciferase report vector carrying a disintegrin and metalloprotease 10(adam10) gene promoter,to screen its stable expression cell line and to analyze its activity.Methods The genome DNA of human neuroblastoma SH-SY5Y cells was extracted as the template.The adam10 gene promoter was amplified by PCR and was cloned into luciferase reporter vector pGL4.17.The adam10 gene promoter luciferase reporter vector pGL4.17-adam10 was constructed and transfected in to SH-SY5Y cells(pGL4.17 vector without promotoer as the negative control and pGL4.17 vector with CMV promoter as the positive control).Then the stable expression cell line was screened by G418 and its fluorescence activity was detect after treating with 1 tμmol/L retinoic acid(RA) for 4 d.Results About 438 bp adam10 gene promoter was successfully amplified by PCR.The pGL4.17-adam10 vector was correct by pCR and double enzyme digestion identification.The cell line stably expressing adam10 gene promoter was obtained after transfecting SH-SY5Y cells by this vector and screening by G418,which had stronger transcriptional activity by detection;1 μmol/L RA could induce high efficiency expression of adam10 gene promoter.Conclusion Human adam10 gene promoter luciferase vector is successfully constructed.adam10 gene promoter can be stably expressed in SH-SY5Y cells,which provides a basis for deeply studying adam10 gene expression regulation,polymorphism analysis and high-throughput drug screening.

Objective:To investigate the people's cardiopulmonary resuscitation (CPR) ability in Hunan Province and whether there are differences in the skill level of CPR among respondents of different ages and education levels.Methods:A self-made questionnaire was conducted to survey people in Hunan Province by means of WeChat from May 2018 to April 2019. The questionnaire referred to 2016 National consensus on cardiopulmonary resuscitation in China issued by Cardiopulmonary Resuscitation Specialized Committee of Chinese Research Hospital Association and 2017 version of American Heart Association (AHA) CPR operational guide, and combined with the CPR assessment scale of Hunan Provincial People's Hospital. The contents of the survey included the basic information of the subjects, the common knowledge and skills of first aid, the willingness to learn CPR skills and implement CPR, and the operational requirements of high quality CPR, etc. Results:A total of 6 563 people received the on-site first aid knowledge questionnaire, and 4 355 people completed and submitted the questionnaire. The recovery rate was 66.36%, of which 3 602 valid questionnaires were from IP in Hunan Province, and the qualified rate was 82.71%. Among the subjects, 1 532 were men (42.53%) and 2 070 were women (57.47%). The majority aged group from 19 to 30 (59.41%) and from 31 to 50 (36.70%). 307 were rural (8.52%) and 3 295 were urban (91.48%); and the majority levels of education were senior or technical secondary school (38.26%) and undergraduate or junior college (44.50%). Of the 3 602 respondents, 39.09% indicated that they had been exposed to CPR knowledge and only 0.36% indicated that they did not wish to participate in CPR training. 69.93% of the respondents said they would actively to help if they wet cardic arrest, and 97.92% of those were willing to do so if the patients were relatives or friends. The survey results showed that only 8.91% of respondents chose relatively high-quality CPR options (chest compressions were performed first, the pressing point was located in the sternum of the midpoint of bilateral nipple line, the pressing frequency was 100-120 times/min, pressing depth was 5-6 cm, the ratio of chest compression to artificial ventilation was 30∶2). Among these people, the correct rate was higher in the ages of 31-50 years old and ≤ 18, 19-30 than those ≥51 years old (12.71% vs. 0%, 6.87%, 8.70%, χ2 = 41.420, P < 0.01). The correct rate of education at graduate level and above was higher than those in junior middle school and below, high school or technical secondary school, undergraduate or junior college (19.57% vs. 2.07%, 3.41%, 13.72%, χ2 = 152.262, P < 0.01). Conclusions:The public in Hunan Province have a strong sense of first aid, and some of the theoretical knowledge of CPR. People between 31 years old and 50 years old of age and with graduate education and above know more about CPR, but the overall mastery of CPR skills is poor. It is necessary to further improve the ability of the public as the first witness by teaching various skills of CPR in various forms.