Research and exploration of salivary biological markers for hepatitis B-related hepatocellular carcinoma / 中华肝脏病杂志

Objective: To study using isotope-labeled relative and absolute quantitative proteomics methodologies to screen for salivary biological markers as a simple, non-invasive tool for identifying hepatitis B-related HCC at an early stage. Methods: Saliva samples were collected to extract salivary proteins. Isotope-labeled relative and absolute quantitative proteomics were used to analyze the differentially expressed proteins between the hepatocellular carcinoma (HCC) and non-HCC groups. Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assays were used to verify differential proteins and identify markers in liver cancer tissues and saliva. Statistical analysis was used to analyze the diagnostic efficiency of salivary biomarkers. Results: 152 differentially expressed salivary proteins were screened out between the HCC and non-HCC groups. Western blot, immunohistochemistry, and enzyme-linked immunosorbent assays validated that the expressions of α-1-acid glycoprotein 1 (ORM1) and alpha-fetoprotein (AFP) were significantly increased in HCC (P < 0.05). There was a significant correlation between salivary AFP and serum AFP (P < 0.05). HCC was diagnosed when salivary α-1-acid glycoprotein 1 combined with AFP. The area under the receiver operating characteristic curve was 0.8726 (95% confidence interval: 0.8104 ~ 0.9347), the sensitivity was 78.3%, and the specificity was 88%. Conclusion: Salivary AFP and α-1-acid glycoprotein 1 can serve as potential biomarkers for hepatitis B-related hepatocellular carcinoma.

Clinical Research of Qingkailing Injection in Adjuvant Treatment of Sepsis and Heat Syndrome in Children / 中国实验方剂学杂志

Objective:To evaluate the clinical efficacy of Qingkailing injection in the treatment of children with sepsis and heat syndrome， and investigate its anti-inflammatory， anticoagulant and protective effects. Method:Eighty patients were randomly divided into control group and observation group with forty cases in each group according to the number table. Both groups received comprehensive treatment measures such as fluid resuscitation， anti-infection， anti-inflammatory， anticoagulation， vasoactive drugs， and protection of vital organ functions. While patients in observation group additionally took Qingkailing injection， 5-10 mL each time， intravenous drip after dilution， 1 time/day. Treatment course was five days in both groups. Before and after treatment， the scores of quick sequential organ failure assessment （qSOFA）， pediatric critical illness score （PCIS） and acute physiology and chronic health evalution Ⅱ （APACHEⅡ） were graded； procalcitonin （PCT）， serum amyloid A protein （SAA） before and after treatment， heparin-binding protein （HBP）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， high-sensitivity C-reactive protein （hs-CRP）， interleukin-6 （IL-6）， IL-10， N-terminal brain sodium Peptide precursor （NT-proBNP）， high-sensitivity cardiac troponin T （hs-cTnT） level， cardiac troponin I （cTnI）， creatine kinase isoenzyme （CK-MB）， <italic>D</italic>-dimer （<italic>D-</italic>D ）， fibrinogen （FIB） and antithrombin Ⅲ （AT-Ⅲ） levels were detected. Result:The APACHEⅡ and qSOFA scores in the observation group were lower than those in the control group （<italic>P</italic><0.05）， while the PCIS score was higher than that in the control group （<italic>P</italic><0.05）. The levels of PCT， SAA，HBP，TNF-<italic>α</italic>， hs-CRP and IL-6 in the observation group were lower than those in the control group （<italic>P</italic><0.01）， while the IL-10 level was higher than that in the control group （<italic>P</italic><0.01）. The levels of NT-proBNP， hs-cTnT， cTnI， CK-MB， <italic>D</italic>-D and FIB in the observation group were lower than those in the control group （<italic>P</italic><0.01）， while the AT-Ⅲ activity was higher than that in the control group （<italic>P</italic><0.01）. Conclusion:Qingkailing injection as the adjuvant therapy in children with sepsis and fever syndrome， can play the role of anti-inflammatory， anticoagulant， reducing infection and myocardial damage， thereby reducing the severity of the disease and improving the prognosis.

Preliminary study of bone marrow mesenchymal stem cell-derived exo-somes in repair of spinal cord injury in rats / 中国病理生理杂志

AIM:To study the influence of bone marrow mesenchymol stem cell-drived exosomes(BMSC-exo-somes)on hindlimb activity,and the numbers of reactive astrocytes and residual neurons in spinal cord injury(SCI)rats. METHODS:BMSCs were cultured using the whole bone marrow adherent culture method and surface markers CD 90 and CD34 were verified by flow cytometry.Exosomes were isolated by ultracentrifugation and the morphology of exosomes was observed under transmission electron microscope.The protein markers CD63 and CD9 were verified by Western blot.After exosomes were applied to SCI rats,the Basso,Beattie and Bresnahan locomotor rating scale score,the Nissl staining of the lesion site,and the numbers of reactive astrocytes and residual neurons were assessed at various time points.RESULTS:Transmission electron microscopic observation revealed the presence of saucer -shaped vesicles.BMSC-exosomes were found to express high levels of CD63 and CD9.Compared with injury group,significant improvement of hindlimb activity scores from day 14 after injury in treatment group was observed(P<0.05),and less reactive astrocytes and more residual neu-rons from day 7 after injury were also observed(P<0.05).CONCLUSION:BMSC-exosomes inhibit reactive astrocytes and death of neurons,and improve hindlimb activity in the rats after SCI.

Diagnostic value of Blumensaat angle for anterior cruciate ligament injury / 中国骨伤

<p><b>OBJECTIVE</b>The receiver operator characteristic(ROC) curve was used to determine the best Blumensaat angle for the diagnosis of anterior cruciate ligament injury, so as to objectively evaluate the diagnostic value of Blumensaat angle for anterior cruciate ligament injury.</p><p><b>METHODS</b>Total 167 patients who had knee arthroscopic treatment in a hospital from January 2015 to January 2016 were retrospectively studied, and the patients' age, gender, left and right limb condition were recorded. The patients were divided into two groups according to Blumensaat angle measured on the MRI: group A(Blumensaat angle<=0°), and group B(Blumensaat angle >0°). The ROC curve was drawn from the statistical data of the group B to get the best critical value of the anterior cruciate ligament injury when the Blumensaat angle was more than 0°. According to the best critical value obtained by ROC curve, the coincidence rate of the total sample was obtained.</p><p><b>RESULTS</b>There were no significant differences in patients' age, gender, and affected limbs. There were 51 patients in group A, in which 49 patients were diagnosed as anterior cruciate injury under arthroscopy(gold standard for diagnosis of anterior cruciate ligament injury), and 2 patients were diagnosed as no anterior cruciate injury under arthroscopy. When the Blumensaat angle was<0°, the coincidence rate was 96.07%. In group B(116 cases), the area under the ROC curve was 0.910, the best cutoff corresponds to 15°; the sensitivity was 70.0%, and the specificity was 95.8%. When the Blumensaat angle was >=15°, the probability of anterior cruciate ligament injury was greater. When the Blumensaat angle was 0° to 15°, the anterior cruciate ligament was more likely to be not injured. The Blumensaat angle<=0°or>=15° were used to diagnose the injury of anterior cruciate ligament. Compared with the results of arthroscopy, the coincidence rate of the total sample was 92.8%.</p><p><b>CONCLUSIONS</b>Blumensaat angle is helpful to diagnose the ACL injuries. When the Blumensaat angle was <=0° or >=15°, the probability of ACL injury is greater.</p>

Correlation analysis of serum calcium levels and risks of diabetes mellitus in middle-aged and elderly men / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the correlation of serum calcium levels and the risks of diabetes mellitus (DM) in middle-aged elderly men.</p><p><b>METHODS</b>A total of 3386 male subjects aged 40-79 years were selected from two local communities of Beijing using stratified random sampling for this cross-sectional survey. The survey was conducted using questionnaires, and all the subjects underwent physical examination and blood tests of serum calcium, vitamin D and other biochemical parameters. The subjects were divided into normal glucose group, prediabetic group and diabetic group to compare their blood biochemical parameters and analyze the risk factors of diabetes.</p><p><b>RESULTS</b>The prevalence of diabetes and pre-diabetes was 32.31% in the 3386 subjects surveyed. Serum calcium, vitamin D and Creatinine (Cr) levels were significantly higher in pre-diabetic and diabetic groups than in the non-diabetic group (P/0.05), and age, body mass index, serum calcium, vitamin D, and Cr levels differed significantly between the former two groups (P/0.05). The prevalence of diabetes increased significantly with serum calcium level (P/0.05). Logistic regression analysis showed that age, body mass index, serum calcium, and serum vitamin D levels were independent risk factors for diabetes (P/0.05).</p><p><b>CONCLUSION</b>Male diabetic patients have abnormal serum calcium levels. An increased age and increased serum calcium levels are associated with higher risks of diabetes, and age, serum calcium, serum vitamin D, Cr, and body mass index are all risk factors of diabetes in men.</p>

Correlation of sex hormones and parathyroid hormone with biochemical markers of bone turnover in aged men / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of serum sex hormones and parathyroid hormone (PTH) with the biochemical markers of bone turnover in aged men.</p><p><b>METHODS</b>We collected the laboratory data of 465 men aged 60- 93 (73. 1 +/- 8. 3) years old, who came for routine physical examinations in our hospital. We obtained the levels of serum follicle- stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), testosterone (T), sex hormone binding globulin (SHBG), PTH, 25-hydroxy-vitamin D3 (25(OH) D3), and bone turnover markers C-terminal telopeptide of type I collagen (CTX), osteocalcin (OC) and amino-terminal propeptide of type I procollagen (PINP). We also determined free testosterone (FT) , bioactive testosterone (BT) , testosterone secretion index (TSI) and FT index (FTI), and analyzed the correlation of each index with the biochemical markers of bone turnover.</p><p><b>RESULTS</b>The concentrations of serum FSH, LH, and SHBG increased, while the levels of FT, BT, TSI, FTI, PTH, CTX, OC and PINP decreased with age, especially in those over 80 years old (P <0.05). PTH was positively correlated with CTX, OC and PINP (r =0. 227, 0. 269 and 0. 162, P <0. 01), even after the adjustment for age, while SHBG negatively correlated with OC (r = -0. 100, P <0.05). The bone turnover markers increased with the elevation of the PTH quartiles, with significant differences between the first and the fourth quartile (P <0. 01). Multiple stepwise regression analysis showed that age was correlated inversely with CTX, OC and PINP ( beta = -0. 126, -0. 141 and -0. 122, P <0.05) , PTH positively with the three markers (beta = 0. 196, 0.279 and 0.189; P <0. 001), and SHBG negatively with OC ( beta = -0. 100, P <0.05) .</p><p><b>CONCLUSION</b>Aging is the fundamental cause of reduced bone turnover in aged men. The levels serum PTH and SHBG are significantly associated with the biochemical markers of bone turnover.</p>

Risk factors analysis of nonalcoholic fatty liver disease in Chinese men / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore risk factors of nonalcoholic fatty liver disease (NAFLD) in men in order to provide a theoretical basis for developing more effective NAFLD prevention and control strategies.</p><p><b>METHODS</b>One-hundred-and-two male patients (37.3+/-11.4 years old) hospitalized with NAFLD at the Dongnan Affiliated Hospital of Xiamen University between January 2009 and December 2010 were enrolled in the study, along with 23 age-matched healthy men (34.4+/-16.7 years old) to serve as the control group. The correlation(s) of body mass index (BMI; overweight defined as more than or equal to 22.717 kg/m2), waist circumference (WC), waist-to-hip ratio (WHR; central obesity defined as more than or equal to 0.866), fasting plasma glucose (FPG), triglyceride (TG), and total cholesterol (TC) with NAFLD was analyzed by univariate and multivariate logistic regression analyses. Receiver operating characteristic (ROC) curves were used to select proper thresholds for classification.</p><p><b>RESULTS</b>BMI, WC, WHR, FPG, TG, and TC were significantly different between the cases and controls (P less than 0.01). BMI, WC, WHR, TG and TC were identified as risk factors of NAFLD in these male cases (P less than 0.01). Relative to WC, TG and TC, both BMI and WHR had significant predictive value for NAFLD (odds ratio (OR) = 10.819 and 10.588, respectively). In addition, BMI had the highest diagnostic value for the prediction of NAFLD (area under the curve (AUC) = 0.931) followed by WHR (AUC = 0.879).</p><p><b>CONCLUSION</b>BMI, WC, WHR, TG, and TC are risk factors of NAFLD in Chinese men. BMI and WHR are effective anthroposomatology indices of NAFLD and may be useful factors on which to base future prevention and early diagnosis strategies for NAFLD in males.</p>

Relationship between mannose-binding protein gene polymorphisms and disease progression and HBV DNA in patients with chronic HBV infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To determine the influences of Mannose binding protein (MBP) gene polymorphisms on HBV DNA loads and on the progression of liver disease in patients with chronic HBV infection.</p><p><b>METHOD</b>The Codons on 54 MBP gene polymorphisms and HBV DNA loads in a cohort of 395 patients with chronic HBV infection, including 244 with chronic hepatitis B (CHB), 151 with liver cirrhosis (LC) and 88 normal controls were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and fluorescent quantitative PCR (FQ-PCR).</p><p><b>RESULT</b>The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC in CHB group showed no significant differences comparing to the normal control group (P > 0.05). The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC on CHB group (severe), compensation phase of LC group and decompensation phase of LC group were higher than those in the normal control group (P < 0.05), the genetic polymorphism of decompensation of LC was 36.5%, highest of all. The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC of patients with chronic HBV infection were not changed with the differences of HBV-DNA loads.</p><p><b>CONCLUSION</b>The codes on 54 MBP gene polymorphisms is not closely related to HBV DNA loads, but was associated with the progression of hepatitis B infection.</p>

The associated research on the expression of RUNX3 mRNA and protein in hepatic cell carcinoma as well as the relationship with clinical pathological parameters / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the expression of RUNX3 mRNA and protein in hepatic cell carcinoma (HCC) and surrounding normal tissue, to analyze the relationship between RUNX3 expression and clinical pathological parameters.</p><p><b>METHODS</b>Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) were performed to detect the expression of RUNX3 mRNA and protein in HCC and surrounding normal tissue respectively, and their relationship with clinical pathological parameters were analyzed.</p><p><b>RESULTS</b>The relative expression value of RUNX3 mRNA found in 51 cases HCC was 0.4509 +/- 0.0963, and that did in 51 cases surrounding normal tissue was 0.9147 +/- 0.0222. The difference of RUNX3 mRNA expression between two kinds of samples was statistically significant (t = 33.6087, P < 0.001). The positives rate of RUNX3 protein expression found in 51 cases HCC tissue was 49.02% (25/51) and that did in 51 cases surrounding normal tissue was 82.35% (42/51). The difference of RUNX3 protein expression between two kinds of samples was statistically significant (chi2 = 12.5706, P < 0.005). The difference of RUNX3 mRNA and protein expression in some clinical pathological parameters involving differentiation degree, invasion, cancer thrombus and diversion in liver were statistically significant (P < 0.05). However that were not in another clinical pathological parameters involving gender, cancer diameter, cancer location as well as hemorrhage and necrosis of cancer, histotype (P > 0.05).</p><p><b>CONCLUSION</b>The expression of RUNX3 mRNA and protein in HCC were significantly lower than that in surrounding normal tissue. The lower expression of runx3 protein in the HCC probably plays an important role in the tumorigenesis and development of HCC. The RUNX3 gene may be an anti-oncogene of HCC.</p>

Ultrastructure and electrophysiology of astrocytes differentiated from adult adipose-derived stromal cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Adipose-derived stromal cell (ADSC) differentiation into neural cells in vitro is becoming widely studied. However, there are few reports on astrocytes following differentiation, and particularly on maturation and electrophysiology. In this study, we used various methods to determine ADSC-derived astrocyte maturity.</p><p><b>METHODS</b>Chemical induction with isobutylmethylxanthine (IBMX) was used to differentiate adult ADSCs into astrocytes followed by hematoxylin-eosin (HE) staining to observe morphology and transmission electron microscopy for cellular ultrastructure assessment. Immunofluorescence was used to detect expression of neural stem cell marker nestin as well as glial markers glial fibrillary acidic protein (GFAP) and S-100. In addition, we measured membrane potentials in bis-(1,3-dibarbituric acid) trimethine oxanol-labeled ADSCs and astrocytes by stimulation with a high potassium solution under an inverted fluorescence microscope. Finally, cell cycle distribution was detected by flow cytometry.</p><p><b>RESULTS</b>Typical astrocyte morphology was shown by HE staining after 48-hour differentiation. Glial fibril was observed with transmission electron microscopy. GFAP and S-100 were not expressed in the control group, but were expressed within 24-hour differentiation and reached a maximum at day 14 with no change up to day 28. Nestin was weakly expressed in control cells and also reached a maximum at day 14 with the percentage of positive cells constant until day 21 followed by a decrease. Differentiated cell membrane potentials after stimulation with potassium were slightly increased, and then gradually declined over time. There was no significant membrane potential change in the control group. Flow cytometry showed that the percentage of cells in G0/G1 phase was 93% and only 5% in S phase.</p><p><b>CONCLUSION</b>ADSCs were differentiated into mature astrocytes with typical characteristics including morphology, ultrastructure, marker protein expression, mature potassium channels and mitotic capacity.</p>

Expression of RUNX3 protein in hepatic cell carcinoma and its clinicopathological implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of RUNX3 protein in hepatic cell carcinoma (HCC) and its relationship with the clinicopathological factors of HCC.</p><p><b>METHODS</b>Immunohistochemistry was performed to detect the expression of RUNX3 protein in HCC and the surrounding normal tissues, and the relation of RUNX3 with the clinicopathological factors of HCC was analyzed.</p><p><b>RESULTS</b>The positivity rate of RUNX3 protein expression was 49.02% (25/51) in HCC tissues, significantly lower than that in the surrounding normal tissues [82.35% (42/51), Χ(2)=12.5706, P<0.01). RUNX3 protein expression varied significantly with such pathological factors as the differentiation degree, cancer-associated thrombosis in the portal vein and intrahepatic metastasis (P<0.05), but not with tumor diameter, location, tumoral hemorrhage, necrosis or histotypes of the tumor (P>0.05).</p><p><b>CONCLUSION</b>RUNX3 protein expression is lowered in HCC as compared with that in the surrounding normal tissue, suggesting an important role of RUNX3 in the tumorigenesis and development of HCC and the possible identity of RUNX3 gene as an anti-oncogene of HCC.</p>

Factor analysis of impaired glucose regulation reversing to normal glycemia following one year's health education / 中华内分泌代谢杂志

The outcome and influencing factors in the reversion of impaired glucose regulation(IGR)to normal glycemia(NG)after health education for one year were analyzed by the criterion of American Diabetes Association 2003.The results showed that the improvement of glucose regulation well accorded with the improvement of insulin resistance and islet 13-cell function.Fasting plasma glucose,fasting insulin,triglycerides, insulin resistance and islet?-cell function were the influencing factors for the reversion of IGR to NG.