ObjectiveTo investigate the molecular mechanism by which Si Junzitang in intervening in the development of hepatocellular carcinoma （HCC） by regulating the O-linked β-N-acetylglucosamine modification （O-GlcNAcylation） of nuclear factor kappa-B （NF-κB） p65 in the paracancerous tissues. MethodThe orthotopic liver cancer mouse model was established. Twenty-four C57BL/6 mice were randomly divided into four groups： Normal group， model group， Si Junzitang low-dose group （10 g·kg^-1）， and Si Junzitang high-dose group （25 g·kg^-1）， with 6 mice in each group. The O-GlcNAcylation level and phosphorylation modification level of p65 in the paracancerous tissues were detected using Western blot. The O-GlcNAcylation of p65 was assessed using immunoprecipitation （IP）. The mRNA expression of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， transforming growth factor-β₁ （TGF-β₁）， vascular endothelial growth factor A （VEGFA）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9 （MMP-9） in the paracancerous tissues was detected using real-time quantitative polymerase chain reaction （Real-time PCR）. The tumor number， liver weight， locomotor activity， grip strength， and Qi status of the mice were observed and analyzed. ResultCompared with the normal group， the model group showed a significant decrease in O-GlcNAcylation in the paracancerous tissues （P<0.01）， a significant decrease in p65 O-GlcNAcylation （P<0.01）， a significant increase in p65 phosphorylation （P<0.01）， significantly elevated mRNA levels of cytokines IL-6， TNF-α， TGF-β₁， VEGFA， MMP-2， and MMP-9 （P<0.01）， significantly increased liver weight （P<0.01）， significantly declined grip strength， number of grid crossings， and number of vertical stand-ups （P<0.01）， and significantly dwindled Qi status （P<0.01）. Compared with model group， the Si Junzitang low-dose and high-dose groups showed significantly increased levels of O-GlcNAcylation in the paracancerous tissues （P<0.05， P<0.01）， significantly upregulated p65 O-GlcNAcylation levels （P<0.05， P<0.01）， and significantly decreased p65 phosphorylation levels （P<0.01）. In the Si Junzitang low-dose group， the mRNA levels of IL-6， TGF-β₁， and VEGFA significantly decreased （P<0.05， P<0.01）. In the Si Junzitang high-dose group， the mRNA levels of IL-6， TNF-α， TGF-β₁， VEGFA， MMP-2， and MMP-9 significantly decreased （P<0.01）， the number of tumors larger than 3 mm in diameter significantly decreased （P<0.01）， and liver weight significantly decreased （P<0.05）. Additionally， grip strength， number of grid crossings， and number of vertical stand-ups significantly increased （P<0.05， P<0.01）， along with a significant increase in qi status （P<0.01）. ConclusionSi Junzitang can inhibit the progression of orthotopic HCC in mice， which may be achieved by increasing the O-GlcNAcylation level in the paracancerous tissues， enhancing the O-GlcNAcylation of p65， inhibiting the phosphorylation modification of p65， and ultimately suppressing the expression of downstream IL-6， TNF-α， TGF-β₁， VEGFA， MMP-2， and MMP-9.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Researches on detection of human papillomavirus (HPV) high-risk samples were carried out by poly-merase chain reaction (PCR) coupled with microchip electrophoresis (MCE). Herein, we introduced a simple, rapid, automated method for detecting high-risk samples HPV16 and HPV18. In this research, general primers were initially selected to obtain sufficient detectable yield by PCR to verify feasibility of MCM method for HPV detection, then type-specific primers were further used to evaluate the specificity of MCE method. The results indicated MCE method was capable of specifically detecting high-risk HPV16 and HPV18, and also enabled simultaneous detection of multiplex samples. This MCE method described here has been successfully applied to HPV detection and displayed excellent reliability demonstrating by sequencing results. The inherent capability of MCE facilitated HPV detection conducted in a small chip with automated, high throughput, massive parallelized analysis. We envision that MCE method will definitely pave a way for clinical diagnosis, and even on-site screening of cervical cancer.

Objective To investigate the prevalence and correlates of depression in residents aged 15 and older in Hainan province.Methods Stratified multistage random sampling methods were utilized to identify 12 117 individuals (≥15 years old) from 59 villages (neighborhoods) in 24 towns (streets) within 6 counties (cities) in Hainan Province from September 2010 to November 2011.The subjects were screened with an expanded version of the General Health Questionnaire (GHQ-12) and the respondents were divided into three groups by high risk,moderate risk and low risk of mental disorder,followed by formal diagnosis according to the Structured Clinical Interview for DSM-IV-TR (SCID-I/P) by psychiatrists among 100％ subjects in group with high risk,40％ subjects in group with moderate risk and 10％ subjects in group with low risk.The adjusted rate,standardized rate,and 95％ confidence interval (95％CI) of the one-month and lifetime prevalence were also calculated among the 3 groups of individuals who were diagnosed with depressive disorder.The difference in whether they suffered depression was determined by the chi-squared test.Correlations to depressive disorder and the odds ratio (OR) were explored with multiple regression analysis.Results There were 97 cases of depressive disorders (1-month) and 166 cases of lifetime depressive disorders finally confirmed.The adjusted rate of 1-month prevalence was 1.38％ (95％CI:1.12-1.59) and the life-time adjusted prevalence rate was 2.80％ (95％CI:2.51-3.09).Female (OR=1.55,95％CI:1.12-2.14),mid-age(50-64 y,OR=1.84,95％CI:1.13-2.99),being divorced or living separated (OR=4.87,95％CI:1.86-12.73),suffering chronic diseases (OR=2.19,95％CI:1.56-3.07),and low family income were significantly associated with suffering depressive disorder.Conclusions The prevalence of the depressive disorder among residents aged 15 and older in Hainan province is lower than the nationwide prevalence.People who are female,mid-age,being divorced or not living with the partner,earning a low income,and suffering chronic diseases have more predisposition to develop the depressive disorder in Hainan province.

Objective To investigate the prevalence and correlates of depression in residents aged 15 and older in Hainan province.Methods Stratified multistage random sampling methods were utilized to identify 12 117 individuals (≥15 years old) from 59 villages (neighborhoods) in 24 towns (streets) within 6 counties (cities) in Hainan Province from September 2010 to November 2011.The subjects were screened with an expanded version of the General Health Questionnaire (GHQ-12) and the respondents were divided into three groups by high risk,moderate risk and low risk of mental disorder,followed by formal diagnosis according to the Structured Clinical Interview for DSM-IV-TR (SCID-I/P) by psychiatrists among 100％ subjects in group with high risk,40％ subjects in group with moderate risk and 10％ subjects in group with low risk.The adjusted rate,standardized rate,and 95％ confidence interval (95％CI) of the one-month and lifetime prevalence were also calculated among the 3 groups of individuals who were diagnosed with depressive disorder.The difference in whether they suffered depression was determined by the chi-squared test.Correlations to depressive disorder and the odds ratio (OR) were explored with multiple regression analysis.Results There were 97 cases of depressive disorders (1-month) and 166 cases of lifetime depressive disorders finally confirmed.The adjusted rate of 1-month prevalence was 1.38％ (95％CI:1.12-1.59) and the life-time adjusted prevalence rate was 2.80％ (95％CI:2.51-3.09).Female (OR=1.55,95％CI:1.12-2.14),mid-age(50-64 y,OR=1.84,95％CI:1.13-2.99),being divorced or living separated (OR=4.87,95％CI:1.86-12.73),suffering chronic diseases (OR=2.19,95％CI:1.56-3.07),and low family income were significantly associated with suffering depressive disorder.Conclusions The prevalence of the depressive disorder among residents aged 15 and older in Hainan province is lower than the nationwide prevalence.People who are female,mid-age,being divorced or not living with the partner,earning a low income,and suffering chronic diseases have more predisposition to develop the depressive disorder in Hainan province.

Objective To investigate the changes of the levels of sonic hedgehog (SHH) and vascular endothelial growth factor (VEGF) in serum and its relationship with collateral circulation in patients with symptomatic middle cerebral artery stenosis. Methods From January 2015 to January 2018, a total of 268 patients with acute ischemic stroke confirmed as unilateral middle cerebral artery M1 segment (MCA-M1) severe stenosis or occlusion by digital subtract angiography (DSA) were enrolled. The baseline clinical data were collected. According to the establishment of collateral circulation shown by DSA, they were divided into good collateral circulation group (152 patients) and poor collateral circulation group (116 patients). The levels of SHH and VEGF in serum were detected by enzyme linked immunosorbent assay (ELISA), the expression characteristics of SHH and VEGF in serum and the relative factors influencing the establishment of collateral circulation were analyzed. Results The levels of serum SHH and VEGF in good collateral circulation group were significantly higher than those in poor collateral circulation group (P < 0.01). Pearson correlation analysis showed that there was a positive correlation between SHH and VEGF (r=0.758, P < 0.01). Multivariate Logistic regression analysis showed that the levels of serum SHH ( OR=0.310, 95% CI 0.117-0.819, P=0.018) and VEGF ( OR=0.361, 95% CI 0.147-0.887, P=0.026) were independent protective factors for the establishment of collateral circulation. Diabetes ( OR=3.094, 95% CI 1.321-7.245, P=0.009) was independent risk factor for the establishment of collateral circulation. Conclusions The levels of serum SHH and VEGF are closely related to the formation of collateral circulation and they are independent protective factors. SHH may be involved in the establishment of cerebral collateral circulation by regulating the expression of VEGF and diabetes is not conducive to the formation of collateral circulation.

Objective To evaluate the effects of baseline and changes of peritoneal transport characteristics on the prognosis of maintaining peritoneal dialysis (PD) patients.Methods Five hundred and eight-six PD patients who started PD from September 11,2006 to October 30,2014 in a single center were included and followed up until March 30,2016.According to their baseline D/Pcr value in peritoneal equilibrium test (PET),the patients were divided into high transport (H) group (D/ Pcr 0.82-1.03),high average transport (HA) group (D/Pcr 0.65-0.81),low average transport (LA) group (D/Pcr 0.50-0.64) and low transport (L) group (D/Pcr 0.34-0.49).According to the changes of follow-up D/Pcr comparing with baseline D/Pcr,the patients were also divided into ascending group,descending group and no-change group.The patient and technical survival rates were estimated by Kaplan-Meier analysis.Cox proportional hazards analyses were used to analyze the risk factors for PD patient death and technical failure.Results There were 67 patients in L group,229 patients in LA group,252 patients in HA group,and 38 patients in H group.The patient survival rate in H group was significantly lower than those of L group (P=0.036),LA group (P=0.008) and HA group (P=0.041).There was no significant difference on technical survival rate among these 4 groups.According to the tendency of follow-up D/Pcr changes,there were 127 patients in ascending group,101 patients in descending group and 179 patients in no-change group.There was no significant difference on patient survival among these 3 groups (P=0.064).However in patients with a high transport rate (D/Pcr≥0.65),the patient survival was lower in descending group than those in ascending group (P=0.033) and nochange group (P=0.049).Age over 65 years old (HR=2.499),malnutrition during follow-up (HR=3.144),ultrafiltration less than 400 ml/d during follow-up (HR=1.863) and high sensitive C reactive protein≥ 10 mg/L (HR=4.526) were the independent risk factors for patient death (all P ＜ 0.05).Gender (HR=1.609),age over 65 years old (HR=1.929),ultrafiltration less than 400 ml/d during follow-up (HR=1.708),high sensitive C reactive protein ≥10 mg/L (HR=1.829),malnutrition (HR=1.876) and change of peritoneal transport function (HR=0.579) affect technical failure (all P ＜ 0.05).Conclusions The survival rate of PD patients with basal high peritoneal transit is relatively low,especially for patients with descending transport rate during follow-up.The concern on the peritoneal transport status is constructive for the prognosis of PD patients.

Objective To understand the plasma protein differential expression before and after cardiopulmonary bypass (CPB)through conducting the comparative proteomics study on rats in order to find the plasma markers with potential value in the early diagnosis of CPB resulted complications.Methods 10 adult male SD rats were divided into the experiment group and the con-trol group randomly (n=5),and took food and water freely before operation.The rat models of CPB were constructed in the experi-ment group.But no any CPB operation was administered in the control group in addition to anesthesia induction,arterial and venous puncture procedure.1 mL of blood sample was extracted for separating plasma before CPB and at the end of CPB in the two groups. The total plasma protein was purified.Then the 2-dimensional electrophoresis and the scanning imaging by ImageScanner were per-formed.The protein spots verified to be differential expression were performed the cutting,enzymolysis and peptide fragment ex-traction.Finally the mass spectrometer was adopted to conduct the analysis and identification.Results The number of visualized spots was increased significantly after CPB.17 protein spots with up-regulated expression were identified as differential expression caused by CPB.5 proteins were verified by mass spectrometer analysis and database research.They were gelsolin,haptoglobin,apo-lipoprotein A-1,immunoglobulin gamma-2b and Ba1-647 respectively.Conclusion CPB can cause the differential expression of plas-ma proteins in rat model.According to the function analysis,gelsolin,haptoglobin and apolipoprotein A-1 have the potentiality of be-ing the plasma markers for studying CPB complications.

This article was aimed to study the proteomic spectra expression of traditional Chinese medicine (TCM) cold and heat constitution rats with two-dimensional gel electrophoresis (2-DE), in order to search for the cold and heat-associated proteins for the investigation of the biological basis of TCM cold and heat body constitution formation. The total protein in rat’s liver cell was extracted. The 2-DE and MALDI-TOF/TOF mass spectrometry (MS) were used in the screening and identification of differentially expressed proteins of cold and heat constitution rats. The results showed that a total of 10 different points in the protein expression were obtained with statistical significance after screening and MS, which were carbamoyl-phosphate synthase, protein disulfide isomerase associated 3, catalase, hydroperoxide isomerase, cytosol aminopeptidase, glutamate dehydrogenase 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, heat shock protein 60 (Hsp60) precursor, homocysteine, 78 kDa glucose-regulated protein precursor. It was concluded that some differences were existed in the proteomic spectra expression of TCM cold and heat constitution rats. The abnormality of enzyme protein metabolism may be one of the material bases for the formation of cold and heat constitution.