Construction of SNP-STR Multiplex Amplification System with Genetic Markers and Its Forensic Application / 法医学杂志

Objective To select and develop a SNP-STR multiplex amplification system with genetic markers compatible with current STR databases. To understand its genetic polymorphisms in Sichuan Han population and its application value in DNA mixture analysis. Methods Based on the STR genetic markers in commercial kits, SNPs adjacent to these STR markers were selected to be SNP-STR genetic markers. A SNP-STR multiplex amplification system with genetic markers based on allele-specific amplification was constructed using allele-specific amplification primers. The genetic polymorphism of the system in the Sichuan Han population was investigated and the efficiency of systems with different numbers of loci to detect the two individual DNA mixture samples was evaluated. Results An allele-specific multiplex amplification system constituted of 13 SNP-STR genetic markers was selected and constructed. In Sichuan Han population, the heterozygosity of each locus ranged from 0.76 to 0.88, and the combined discrimination power reached 0.999 999 999 999 999 968. In the analysis of the two individual DNA mixture samples： for single-locus amplification, the genotype of the minor components can still be detected when the mixture ratio reaches 1 000∶1; for multiple loci multiplex amplification, the maximum mixture ratio can reach 500∶1. As the number of loci in the system increased, the detection efficiency of the minor components in the DNA mixture decreased. Conclusion SNP-STR genetic markers have a higher polymorphism than STR. The multiplex amplification system made of SNP-STR genetic markers has a better analysis efficiency for mixed samples than traditional STR multiplex amplification system.

Historical story on natural medicinal chemistry: Rapamycin / 中草药

The study on natural product chemistry plays an important role in drug development and microbe is one of the main sources of new drugs. Rapamycin is isolated from the bacterium ensconced in a sample of Easter Island's soil, the compound has led to several approved immunosuppressant and anticancer drugs. The researchers studying on the target of rapamycin have got the "Nobel-class" achievements. In this paper, the discovery and study on rapamycin is summarized to provide the references for future related research.

Historical story on natural medicinal chemistry: The most preeminent natural organic chemist of 20th century-Robert Burns Woodward / 中草药

Robert Burns Woodward was a well-known natural organic chemist. This paper reviewed the work and his contribution to the development of natural product chemistry and total synthesis of natural products including spectroscopic methods such as UV, IR, and NMR, the Octant Rule, structure elucidation and total synthesis of natural products, theory study, biogenesis, et al. This is one of the series papers about historical story on natural medicinal chemistry, which was dedicated to Robert Burns Woodward on the occasion of his 100th birthday.

Study on the mechanism of amniotic extraction inhibiting haze formation after Epi-LASIK in rabbit cornea / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the probably mechanism of amniotic extraction inhibiting haze formation after epipolis laser in situ keratomileusis (Epi-LASIK) in rabbit cornea.METHODS: Thirty rabbit corneas were performed with Epi-LASIK.All eyes were randomly divided into three groups: eyes treated with amniotic extraction (AE group),eyes treated with 1g/L dexamethasone (hormone group) and eyes treated with solvent (solvent control group).Haze grade evaluation was performed under the slit lamp after Epi-LASIK for 1,4 and 8wk.The repair of corneal epithelium was observed by using HE staining,and the expression of NF-kB protein P65 was detected by immunohistochemistry.The expression levels of inflammatory cytokines (TNF-α,TGF-β1 and IL-1) and anti-inflammatory cytokines (IL-4,IL-10 and IL-13) were determined by ELISA.RESULTS: HE staining showed that the basal cells of corneal epithelium were more uniform and arranged regularly in AE groups after Epi-LASIK for 1wk as compared with the hormone group and the solvent control group.After 4wk,there were a few of new collagen fibers in the superficial stroma of AE group,forming a small amount of scar.After 8wk,the corneal stroma of AE group showed a small amount of new collagen fibers,arranged regularly,and rarely formed scar.At the early stage (1 and 4wk),AE treatment has an obviously effect on inhibiting the secretion of inflammatory factors (TNF-α,TGF-β1 and IL-1) and anti-inflammatory factors (IL-4,IL-10 and IL-13),and the difference was statistically significant (P<0.01).Moreover,the activation of the NF-kB signaling pathway was significantly inhibited by treatment with AE in the early postoperative period (1 and 4wk).CONCLUSION: Amniotic extraction may reduce the inflammatory response in corneal epithelial cells by inhibiting the NF-kB signaling pathway,thereby inhibiting the formation of collagen and scar and the occurrence of haze.

Influence of Leukodeplated Blood Transfusion on Cellular Immunity of Acute Leukemia Patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the influence of leukodeplated blood transfusion on cellular immunity of patients with acute leuemia, so as to provide support for application of leuko-deplated blood transfusion in clinic.</p><p><b>METHODS</b>A total of 100 AL patients from January 2012 to December 2015 were chosen, and were divided into 2 groups: leukodeplated blood transfusion group(50 cases) and routine blood transfusion group(RBT) as control (50 cases). The effective rate, side effects, peripheral blood T cells and expression level of TLR2 and TLR4 were compared between 2 groups.</p><p><b>RESULTS</b>The expression levels CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+) of TLR2 and TLR4 in control group were (52.18±2.14)%, （27.28±1.19）%,（24.21±1.65）%,1.22±0.18,0.62±0.04 and 0.57±0.05, respectively, after treatment; while these indicators in LdBT group were （52.18±2.14）%,（30.97±2.01）%,（27.08±1.55）%,1.39±0.24,0.91±0.06 and 0.87±0.07, respectively, and above-mentioned indicators in LdBT group were significantly higher than those in control group（P<0.05）. Compared with these indicators before treatment, CD3(+), CD4(+), CD8(+) and CD4(+)/CD8(+) in the patients increased significantly（P<0.05）. The efficiency was 92.00% (46/50) in LdBT group, and 84.00% (42/50) in control group, without statistically significant difference（P>0.05）. The rate of side effects in study group was 6% (3/50), 18% (9/50) in control group, with statistically significance difference （P<0.05）.</p><p><b>CONCLUSION</b>Leukodeplated blood transfusion can improve the cellular immunity of AL patients, and reduce the rate of side effects.</p>

Clinical research of the relation of hepatitis B surface antigen (HBsAg) quantification and hepatic tissue pathological staging / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the correlation of serum hepatitis B surface antigen (HBsAg) level and hepatic tissue pathological staging in the chronic hepatitis B infected persons.</p><p><b>METHODS</b>Collect the clinical data of 272 cases who are HBsAg-positive more than 6 months and accepted hepatic biopsy in our hospital. Detect serum HBsAg quantification, ALT, HBV DNA, complete blood count, hepatic tissue pathological staging, grouping the cases according to the stage of inflammation and the fibrosis degree respectively. Observe serum HBsAg quantification, HBV DNA and the stage of inflammation and the fibrosis degree. Analyse the correlation between HBsAg quantification and HBV DNA.</p><p><b>RESULTS</b>The correlation of serum HBsAg level and HBV DNA is notable. Serum HBsAg level is a variable affecting hepatic tissue pathological stage significantly.</p><p><b>CONCLUSIONS</b>Serum HBsAg level is a marker having higher specificity and sensitivity to diagnose the hepatic fibrosis.</p>

Impact of liver steatosis on antiviral effects of pegylated interferon-alpha in patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the impact of hepatic steatosis on virologic response in chronic hepatitis B (CHB) patients treated with pegylated interferon-alpha (PEG-IFNa).</p><p><b>METHODS</b>Ninety-six naive patients positive for hepatitis B e antigen (HBeAg) and with biopsy-proven CHB were administered PEG-IFNa-2a or PEG-IFNa-2b for 48 weeks. Virologic response (HBeAg clearance and hepatitis B virus (HBV) DNA less than 5 log10 copies/ml) and biochemical response (alanine transaminase (ALT) normalization) were compared between patients with (n=34) and without (n=62) steatosis.</p><p><b>RESULTS</b>The HBV DNA titer in the steatosis group was significantly lower than that of the non-steatosis group (6.961.27 vs. 7.541.28 log10 copies/ml; t=2.161, P=0.033). After 48 weeks of PEG-IFNa treatments, there was no significant difference in HBeAg seroconversion or the percentage of undetectable HBV DNA (less than 3 log10 copies/ml) between steatosis and non-steatosis patients. However, the steatosis patients presented with a significantly lower complete response rate (virologic response plus biochemical response) compared to non-steatosis patients (26.5% vs. 48.4%; x² =4.373, P=0.037). Of the 45 CHB patients with undetectable HBV DNA after 48 weeks of treatment, seven did not achieve ALT normalization. The rate of patients with non-biochemical response was significantly higher in the steatosis group than in the non-steatosis group (33.3% vs. 6.67%; P=0.032).</p><p><b>CONCLUSION</b>Hepatic steatosis does not affect the virologic response, but does affect the biochemical response in CHB patients treated with PEG-IFNa for 48 weeks.</p>

Value of delta model for end-stage liver disease in evaluating the prognosis of liver failure patients with hepatitis B virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the prognostic value of the model for end-stage liver disease (MELD) and deltaMELD in liver failure patients infected with hepatitis B virus.</p><p><b>METHODS</b>Based on prospective study design, 98 hospitalized cases were studied and followed up for 24 weeks. The clinical data were recorded. We calculated the score of MELD and deltaMELD, and also compare the score between the survival group and death group. Using ROC curve plotting obtained the better decisive threshold. The case fatality rate were compared at different time points which the patients were classified by the best critical value of MELD and deltaMELD. We draw the Kaplan-Meier survival curve of different group and analyse the change of survival rate by log-rank analysis.</p><p><b>RESULTS</b>52 of 97 patients died and 46 survive during 24 weeks of followup. There was significant difference between the two groups for MELD and deltaMELD (P < 0.01). The case fatality rate in group which MELD > or = 23 was obviously higher than in that MELD < 23. The rate in group which deltaMELD > 4.5 was obviously higher than in that deltaMELD < 4.5 (P < 0.001). The area under curve (AUC) for the twelfth and 24th week's prognosis judgment of deltaMELD (0.823, 0.815) was larger than that of MELD (0.680, 0.684) (P < 0.05). Survival analyses (Kaplan-Meier) indicated that there were significant differences in cumulative survival rates among the groups which were grouped by optimization critical value ( P = 0. 000).</p><p><b>CONCLUSIONS</b>The scoring system of MELD also applied to the forecasting of prognosis for severe hepatitis B patients in China. The accuracy of deltaMELD to predict the prognosis was higher than that of MELD. The combination of MELD and deltaMELD showed good clinical practical value.</p>

The recent research progress of chemistry of marine natural products / 药学学报

Ocean is a unique and excellent resource that provides a diverse array of intriguing natural products. Marine natural products have demonstrated significant and extremely potent biological activities and have captured the attention of natural products chemists in the past few decades. It is increasingly recognized that a wealth of fascinating natural products and novel chemical entities will play a dominant role in the discovery of useful leads for the development of pharmaceutical agents and provide useful probes to lead to breakthroughs in a variety of life-science fields. This article focused on the research progress of chemistry of marine natural products in recent five years.

Analysis of 96 cases with novel H1N1 influenza A / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To review the epidemiologic and clinical characteristics of 96 cases with novel H1N1 influenza A, and improve the diagnosis and treatment level of novel H1N1 influenza A.</p><p><b>METHODS</b>96 cases of novel H1N1 influenza A admitted to the isolation wards from Oct 20 to Sep 23, 2009 were studied. Their epidemiologic, clinical, laboratory, and radiologic characteristics were analyzed.</p><p><b>RESULTS</b>The median age of the 96 patients was 26.52 +/- 10.62 years (range, 5 to 60 years). Sixty-four of the 96 patients had a close contact with novel H1N1 influenza A patients. The main symptoms included fever 100%, cough 86.4% , sore throat 66.6% and myalgia 32.3%.</p><p><b>CONCLUSION</b>The clinical presentation of novel H1N1 infection is largely indistinguishable from that of seasonal influenza. Combines both a symptom complex with the epidemiological investigation and laboratory characteristics can improve the accuracy of diagnosis of novel H1N1 influenza A.</p>

Effect of caspase 9 related signaling molecules on the apoptosis of human vascular endothelial cell induced by homocysteine / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To understand the role of mitochondria associated signaling pathway in the apoptosis of human vascular endothelial cell induced by homocysteine (Hcy).</p><p><b>METHODS</b>The mRNA and protein expression levels of the up-stream signaling molecules of caspase 3, Bcl 2, caspase 9, and cytosolic cytochrome-c, were investigated. The in vitro cultured human umbilical vein endothelial cells with homocysteine at different concentrations were incubated for 24 h. The expressions of Bcl 2 and caspase 9 at mRNA and protein levels were analyzed by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot. Cytochrome-c in cytoplasm was also detected by Western blot.</p><p><b>RESULTS</b>The expression levels of three signaling molecules were all down-regulated by homocysteine at both mRNA and protein levels in a dose-dependent manner.</p><p><b>CONCLUSION</b>Homocysteine could affect the formation of apoptosome through repressing the expression of Bcl 2 gene and release of cytochrome-c from mitochondria. Decreasing of apoptosome could disturb the activation of caspase 9. The results also indicate that the mitochondria pathway is not the major signaling pathway involved in Hcy-induced apoptosis.</p>

Structural survey of airway sensory receptors in the rabbit using confocal microscopy / 生理学报

Information on the morphology of airway receptors is limited. The present study surveys rabbit airway receptors using immunohistochemical and fluorescent labeling to identify their structure with confocal microscopy. Various receptor types were observed to have multiple branches where a parent axon fed several structures. Receptors were located in different layers of the airway, i.e., smooth muscle, lamina propria (submucosa) and the epithelium. Smooth muscle and submucosal receptors were innervated by thick myelinated fibers, while epithelial receptors were supplied by thin-diameter axons. Structures of smooth muscle receptors and some submucosal receptors covered a relatively large area, while epithelial receptors were less extended. In addition, intrapulmonary ganglia were also labeled. Some were closely associated with the axons of smooth muscle receptors.

Increased atria expression of receptor activity-modifying proteins in heart failure patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Receptor activity-modifying proteins (RAMPs) determine the ligand specificity of the calcitonin receptor-like receptor (CRLR); co-expression of RAMP1 and CRLR results in a calcitonin gene related peptide (CGRP) receptor, whereas the association of RAMP2 or RAMP3 with CRLR gives an adrenomedullin(ADM) receptor. As CGRP and ADM may play a beneficial role in heart failure, this study aimed at the question whether RAMPs mRNAs are changed in heart failure.</p><p><b>METHODS</b>Semi-quantitative reverse transcription-PCR (RT-PCR) was used to detect and quantify the mRNAs of RAMP1 and RAMP3 in the atria of heart failing patients.</p><p><b>RESULTS</b>It was found that the expressions of RAMP1, RAMP2 and RAMP3 mRNAs increased with the worsening of heart function, but the expressions of RAMP1 and RAMP2 mRNA decreased at level IV of heart failure.</p><p><b>CONCLUSION</b>The above results demonstrated in the atria of heart failure patients an up-regulation of CGRP receptor by an increase of RAMP1 in association with CRLR and an up-regulation of ADM receptor by an increase of RAMP2 expression in association with CRLR, thus suggesting that CGRP and ADM receptors be playing a functional role in compensating the chronic heart failure in human.</p>

Na(+)/K(+) -ATPase as a marker for detecting pulmonary sensory receptors / 生理学报

Pulmonary vagal afferents provide some of the most important breathing control inputs to the respiratory center. However, the structure of vagal receptors is not yet well known. Lack of an available and effective method to identify pulmonary receptors is the major limiting factor. Here we describe a new approach using confocal microscopy to examine the structure of immunohistochemically stained airway receptors by using Na(+)/K(+)-ATPase as a marker. This method permits detailed observation of pulmonary receptor structures and their relationship to target tissue in the rabbit. The parent axon has multiple branches that supply the receptor structure. Each structure has multiple endings that form leaf-like terminals. This technique permits examining pulmonary receptor structures in detail, and should assist in identifying the morphology of the receptors and the mechanisms of receptor activation.