Application progress of liquid biopsy in the diagnosis and treatment of biliary tract cancer / 中华肝胆外科杂志

With the development of precision medicine and individualized treatment, tissue biopsy in cancer patients diagnosis and therapy has been broadly used. However, because it’s hard to collect enough samples for biliary tract tumors, liquid biopsy was broadly applied for the diagnosis. In liquid biopsy, circulating tumor cells, circulating tumor DNA, and tumor-derived exosomes carrying tumor-specific information are released from tumor tissue into blood, bile, and other body fluids, which makes tumor biopsy samples easily to be obtained in a non-invasive way. At the same time, through a series of morphological and molecular measurements as well as genetic characterization, liquid biopsy can be used to look for the new early diagnostic markers, and therapeutic targets, monitoring progression and prognosis of diseases. This article outlined the current technology used to detect circulating tumor cells, circulating tumor DNA, and tumor-derived exosomes, and summarizes the latest advances in the clinical application of liquid biopsy in biliary tract cancers.

Entecavir add-on Peg-interferon therapy plays a positive role in reversing hepatic fibrosis in treatment-naïve chronic hepatitis B patients: a prospective and randomized controlled trial / 中华医学杂志(英文版)

BACKGROUND@#The efficacy of entecavir (ETV) add-on peg-interferon therapy compared with ETV monotherapy in treatment-naïve hepatitis B virus (HBV) patients remains controversial. We investigated whether adding peg-interferon to ongoing ETV treatment leads to a better curative effect or not.@*METHODS@#All patients have been recruited between August 2013 and January 2015 from the Shanghai Public Health Clinical Center and Zhongshan Hospital (China). Eligible HBV patients (n = 144) were randomly divided (1:1) to receive either ETV monotherapy (n = 70) or peg-interferon add-on therapy from week 26 to 52 (n = 74). Patients were followed-up for at least 2 years. Indexes including hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) seroconversion rate, sustained virologic response, transient elastography value, and histological scores were evaluated every 3 months until the end of the study. The rate of patients with HBsAg loss was defined as the primary endpoint criteria.@*RESULTS@#At week 26, no patient achieved HBsAg seroconversion in either group. At week 52, one patient in the monotherapy group was HBsAg-negative but there was none in the combination therapy group. The monotherapy group showed significantly better liver function recovery results than the combination therapy group. At week 78, one patient in the combination group had HBsAg seroconverted. At week 104, only three patients in the combination therapy group were HBsAg-negative compared with one patient in monotherapy. The mean alanine aminotransferase and aspartate aminotransferase levels and transient elastography values decreased significantly compared with baseline. Both groups showed a favorable decrease in alpha-fetoprotein (monotherapy: 4.5 [2.8, 7.1] vs. 2.2 [1.8, 3.1] ng/mL, P < 0.001; combination therapy: 5.7 [3.0, 18.8] vs. 3.2 [2.0, 4.3] ng/mL, P < 0.001) and an improved result of liver biopsy examination scores. The combination group showed a better improvement in histology compared with the monotherapy group (mean transient elastography value 6.6 [4.9, 9.8] vs. 7.8 [5.4, 11.1] kPa, P = 0.028). But there was no significant difference in HBsAg conversion rate (1.8% [1/56] vs. 4.1% [3/73], P = 0.809) and HBeAg conversion rate (12.5% [7/56] vs. 11.0% [8/73], P = 0.787), as well as HBV-DNA, sustained virologic response (93.2% vs. 98.5%, P = 0.150) between the two groups.@*CONCLUSIONS@#Both therapies supported liver function recovery and histology improvement. Combination therapy did not show better anti-viral efficacy in HBsAg or HBeAg seroconversion compared with monotherapy. However, combination therapy played a more positive role in reversing hepatic fibrosis compared with monotherapy.@*TRIAL REGISTRATION@#ClinicalTrials.gov: NCT02849132; https://clinicaltrials.gov/ct2/show/NCT02849132.

Effect of Angelica sinensis polysaccharide on the osteogenic differentiation of bone marrow mesenchymal stem cells of rats with high glucose levels / 华西口腔医学杂志

OBJECTIVE@#This study aims to evaluate the effect of Angelica sinensis polysaccharide (ASP) on the osteogenic differentiation of the bone marrow mesenchymal stem cells (BMSCs) of rats with high glucose levels.@*METHODS@#Rat BMSCs were isolated and identified by osteogenic and adipogenic differentiation. Then, the BMSCs were divided into three groups as follows: normal control group (5.5 mmol·L⁻¹ glucose), high glucose group (25.5 mmol·L⁻¹ glucose), and ASP+high glucose group (25.5 mmol·L⁻¹ glucose +40 mg·L⁻¹ ASP). The proliferation activities of the BMSCs were detected by CCK8. Alizarin red staining, and alkaline phosphatase activity were used in the examination of osteogenic activity. Quantitative real time-polymerase chain reaction was used to detect the expression levels of the osteogenic genes (Runx2, Osx, OCN, Col-Ⅰ) and the key factors of Wnt/β-catenin signal pathway (CyclinD1, β-catenin). In vivo, a type 2 diabetes rat model was established. The rats were divided into three groups, namely, the normal control group (normal rats), diabetes group (diabetic rats), diabetes+ASP group (diabetic rats, ASP feeding). Then, the tibia bone defect was established. The repair of bone defects in each group was observed through histological examination.@*RESULTS@#The proliferation of BMSCs was higher in the high glucose group and ASP+high glucose group than in the normal control group (P0.05). The number of calcium nodules of BMSCs; alkaline phosphatase activity; and the mRNA expression of Runx2, OCN, Osx, Col-Ⅰ, CyclinD1, β-catenin in the high glucose group were lower than those in the normal control and ASP+high glucose groups (P0.05). The bone mass was significantly lower in the bone defect of the diabetes group than in the bone defect of the normal control or diabetes+ASP group (P0.05).@*CONCLUSIONS@#ASP can promote the osteogenic differentiation of rat BMSCs under high glucose culture and induce bone regeneration in rats with type 2 diabetes. These features may be related to the activation of the Wnt/β-catenin signaling pathway.

Effects of nifedipine on oxidative stress causing by iron-overload in HK-2 cells / 实用医学杂志

Objective To investigate the effect and the mechanism of nifedipine on oxidative stress trip-ping by iron-overload of HK-2 cells. Methods The cells were divided into 4 groups,blank group,iron overload group,nifedipine group and FAC with nifedipine co-treatment group. Cells were treated with FAC or/and nifedipine for 24 hours,and then malonaldehyde (MDA) content,superoxide dismutase (T-SOD) activity,glutathione (GSH) content,intracellular iron content and expression of DMT1 and FPN1 protein were evaluated. Results Compared to the iron overload group,both nifedipine treatment and co-treatment decreased the content of MDA (P < 0.05),increased the activity of SOD(P < 0.05),increased the content of GSH(P < 0.05),reduced the intracellular iron content(P<0.05),increased the expression of DMT1(P<0.05),and increased the expression of FPN1(P < 0.05). Conclusion Nifedipine plays a protective role against oxidative stress induced by iron-overload in HK-2 cells,and it is related to promote DMT1 and FPN1 protein expression and reduce intracellular iron content.

EST-SSR identification, markers development of Ligusticum chuanxiong based on Ligusticum chuanxiong transcriptome sequences / 中国中药杂志

Ligusticum chuanxiong is a well-known traditional Chinese medicine plant. The study on its molecular markers development and germplasm resources is very important. In this study, we obtained 24 422 unigenes by assembling transcriptome sequencing reads of L. chuanxiong root. EST-SSR was detected and 4 073 SSR loci were identified. EST-SSR distribution and characteristic analysis results showed that the mono-nucleotide repeats were the main repeat types, accounting for 41.0%. In addition, the sequences containing SSR were functionally annotated in Gene Ontology (GO) and KEGG pathway and were assigned to 49 GO categories, 242 KEGG pathways, among them 2 201 sequences were annotated against Nr database. By validating 235 EST-SSRs,74 primer pairs were ultimately proved to have high quality amplification. Subsequently, genetic diversity analysis, UPGMA cluster analysis, PCoA analysis and population structure analysis of 34 L. chuanxiong germplasm resources were carried out with 74 primer pairs. In both UPGMA tree and PCoA results, L. chuanxiong resources were clustered into two groups, which are believed to be partial related to their geographical distribution. In this study, EST-SSRs in L. chuanxiong was firstly identified, and newly developed molecular markers would contribute significantly to further genetic diversity study, the purity detection, gene mapping, and molecular breeding.

Influence of coix seed triglyceride combined with transcatheter arterial chemoembolization therapy on the immune function in patients with advanced primary hepatocellular carcinoma / 介入放射学杂志

Objective To investigate the influence of coix seed triglyceride combined with transcatheter arterial chemoembolization (TACE) therapy on AFP, CD4﹢CD25﹢regulatory T (Treg) cells and cellular immune function in patients with advanced primary hepatocellular carcinoma (HCC). Methods A total of 50 patients with inoperable HCC, whose imaging examination showed no distant metastasis, were divided into the study group (n=25) and the control group (n=25). Coix seed triglyceride together with TACE was employed for the patients of the study group, while only TACE was adopted for the patients of the control group. For the patients of the study group, transcatheter hepatic artery infusion of 100 ml coix seed triglyceride was carried out during the performance of TACE, and postoperative intravenous drip of coix seed triglyceride (200 ml/d) was used for 5 days. The peripheral blood samples were collected one week before and one month after the treatment to detect the changes of AFP and T lymphocyte subsets (CD3﹢, CD4﹢, CD8﹢,CD4﹢/CD8﹢ and Treg) levels. One month after the treatment, enhanced CT, MRI or PET-CT was performed to evaluate the necrosis degree of the tumor. Results After the treatment, AFP levels was decreased in both groups, when compared the preoperative data the differences were statistically significant (P0.05). In the study group, the percentage of Treg cells decreased from preoperative (8.27±6.65)%to postoperative (4.22± 1.59)%, the difference was statistically significant (P0.05). Conclusion In treating advanced primary HCC, coix seed triglyceride combined with TACE can reduce the percentage of Treg cells, thus, influence the patient’s cellular immune status and possibly decrease the recurrence rate of HCC after TACE therapy.

The study of the expression of TCR BV CDR3 family in fulminant hepatitis B patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the expression of TCR BV CDR3 family in fulminant hepatitis B (FHB) patients.</p><p><b>METHODS</b>Totally 28 patients with fulminant hepatitis B (FHB) (FHB group), who were treated in our hospital from Oct. 2010. to Mar. 2012, and 20 healthy controls( HC group) were included in the study. PBMCs were isolated from anticogulated blood, and RT-PCR was used to detect the levels of TCR BV CDR3 family in the 2 groups.</p><p><b>RESULTS</b>The levels of DeltaCt1, DeltaCt12 and DeltaCt20 in FHB group were higher than those in HC group (P < 0.05); The levels of DeltaCt5, DeltaCt7, DeltaCt13, DeltaCt14, DeltaCt15, DeltaCt22, DeltaCt23 in FHB group were lower than those in HC group (P < 0.05).</p><p><b>CONCLUSIONS</b>The result indicates that cellular immunology is involved in the pathogenesis of the liver inflammation process of FHB.</p>

A comparative study on the absorption kinetics parameters of rhubarb free anthraquinones between normal dogs and dogs with severe acute pancreatitis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of severe acute pancreatitis (SAP) on absorption kinetic parameters of rhubarb free anthraquinones.</p><p><b>METHODS</b>Eleven healthy Beagle dogs were randomly divided into the normal group (n = 6) and the SAP group (n = 5). The SAP animal model was prepared by surgery through portal vein blood channel building to collect blood from normal dogs and dogs with SAP. The free anthraquinones (20 mg/kg) was given by gastrogavage. The concentrations of five anthraquinones (aloe emodin, rhein, emodin, chrysophanol, and physcion) in the blood plasma of the portal vein and the femoral artery were determined using high performance liquid chromatography (HPLC). The kinetic parameters were calculated using MATLAB2007B Software. The half life (t(1/2Ka)), the absorption peak time (Tmax), the peak concentration (Cmax), the area under the curve [AUC(0-infinity)], and the mean residence time (MRT) were calculated using the statistical moment method. The transport velocity of corresponding medicines from the gastrointestinal tract to the blood (ka) was calculated.</p><p><b>RESULTS</b>There was no difference in the chemical composition absorption type of the portal vein and the femoral artery between the two groups. Aloe emodin could be detected in the portal vein of each animal at each time point, and they were in the quantitative range. Rhein could be detected in the portal vein of each animal at each time point, and they were lower than the quantitative limit at few time points. Emodin and chrysophanol could be detected in the portal vein of partial animals at each time point, and most of them were higher than the quantitative limit. Physcion could be detected only in the portal vein of less animals at few time points. Rhein could be detected in the femoral artery at most time points, but the rhein plasma concentration at most time points were lower than the quantitative limit. Lower concentration of aloe emodin, emodin, and chrysophanol could be detected in the femoral artery at only few time points. Physcion was not detected in the femoral artery. The rhein plasma concentration of the femoral artery and the chrysophanol Cmax of the portal vein at 45 min were higher in the SAP group than in the normal group with statistical difference (P<0.05). There was no statistical difference in the rest indices. The AUC of rhein in the two groups were 59.32% and 66.07% of the total free anthraquinones respectively.</p><p><b>CONCLUSIONS</b>SAP could not obviously affect the absorption kinetics parameters of rhubarb free anthraquinones. The intestinal tract and the liver might possibly play important roles in metabolizing or transforming rhubarb free anthraquinones.</p>

Therapy of aplastic anemia with autologous peripheral mononuclear cells treated by IL-2 and GM-CSF in culture: a long-term follow-up report on 49 patients / 中国实验血液学杂志

This study was purposed to evaluate the long-term outcome and the safety of autologous peripheral blood mononuclear cells (PBMNC) treated by interleukin 2 (IL-2) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the therapy of patients with aplastic anemia (AA). The therapy of 49 patients admitted BG in hospital from April 2001 to December 2007 were analyzed retrospectively. PBMNC were isolated and cultured for 48 hours in presence of IL-2 and GM-CSF. Cells were collected, and 6 × 10(6) - 1 × 10(8) PBMNC were intravenously injected weekly for 4 - 22 months. Hematopoietic recovery was evaluated by examinations of peripheral blood, bone marrow aspirates and bone marrow biopsy. Flow cytometry was used to assess the peripheral T cell subsets before and after treatment. Polymerase chain reaction was performed to observe the clonal diversity of T cell receptor variable β-chain (TCR-Vβ) recombination. The results showed that 37 cases were cured and none of them relapsed during the follow-up, 5 cases were in partial remission, 3 cases got improvement, and 4 cases showed no response. The total efficiency reached up to 91.8%. The ratios of CD4(+)/CD8(+) subsets were abnormal in 39 patients prior to the treatment, and 31 cases restored to the normal range after cell transfusions. Analysis on the clonal diversity of TCR-Vβ recombination in 11 patients showed the transition from monoclonal or biclonal spectratype to polyclonal one. No long-term side effects were documented. It is concluded that the treatment with PBMNC treated by IL-2 and GM-CSF is generally safe and effective. The underlying mechanisms may be in relation to the restoration of cell immunity.

Three cases of eosinophilichyperplastic lymphogranuloma in children's parotid area / 华西口腔医学杂志

From March 2009 to October 2009, three pediatric patients with parotid tumor were cured. Preoperative physical examination showed regional swelling in parotid area, the surface skin was in moderate reddish purple, the border was vague, and the swelling was inactive. The patients' IgE were significantly increased. B ultrasound examination demonstrated the focus was an isoecho with ringlike dark band around, which was concluded as bull's-eye sign. Magnetic resonance imaging (MRI) examination indicated a cystic mass between the skin and parotid. Preoperative diagnosis was eosinophilichyperplastic lymphogranuloma (Kimura's disease) and the granuloma was excised by operation. Pathological examination revealed the capillary vessel hyperplasia in local tissue with a plenty of eosinophils and lymphocytes infiltrating. The disease was confirmed. Although the disease is rare, the diagnosis still could be made by preoperative physical examination, laboratory and imaging examinations.

Clinical experiment of cytokines induced killer cells for treatment of benzene poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To assess the reaction of cytokines induced killer (CIK) cells treatment in hematopoietic injury at different levels on patients with benzene poisoning and seek a novel, safe and effective immunotherapy for benzene poisoning.</p><p><b>METHODS</b>CIK cells were in vitro activated by interleukin-2 (IL-2) and granulocyte-macrophage-colony-stimulating factor (GM-CSF) from the peripheral blood mononuclear cells (PBMC). Thirty-two patients with benzene poisoning were treated with CIK cells. Nineteen patients with mild or moderate benzene poisoning in the control group were treated with VitB4, batilol, leucogen, inosine and stanozolol. The results for treatment of 12 patients with aplastic anemia induced by severe benzene poisoning (the efficacy rate and the case fatality rate) were analyzed. The change of T-lymphocyte subset analyzed by flow cytometry was also observed before and after treatment.</p><p><b>RESULTS</b>For mild or moderate benzene poisoning, the increase of WBC and RLT in CIK group was higher than that in the control group (P < 0.05). The CD(4)/CD(8) levels were significantly increased after CIK treatment. And for severe benzene poisoning, the effective rate of the CIK group was 91.7% and the mortality rate was 0%.</p><p><b>CONCLUSION</b>CIK treatment is safe and effective for hematopoietic injury caused by benzene poisoning. The mechanism may be related with the immune modulation of CIK treatment on immunodeficiency of patients with benzene poisoning.</p>

Advances of forensic entomology in China / 法医学杂志

Forensic entomology is a branch of forensic medicine, which applies studies of insects and arthropods to getting evidence for court and has an analogous advantage in the estimation of the postmortem interval (PMI) and other questions of forensic relevance. The paper expounds its definition and contents and reviews some progress of the studies in some aspects in China such as the constitution and succession of insect community on the different cadavers, the applications of morphological features of insects and the technology of analysis of deoxyribonucleic acid (DNA) in forensic entomology, and forensic entomological toxicology etc.

Measuring volume and density of human organs and its significance in medicine and forensic medicine: lung / 法医学杂志

OBJECT@#To obtain values of normal human lung and diffuse pathological changes, in order to provide a simple and convenient diagnostic method for measuring human visceral organs in the autopsy of pathology and forensic pathology.@*METHODS@#The exact mass, volume and density of normal and pathological lung were synchronously measured with the intelligentized volume-densimeter made by authors.@*RESULTS@#The date-base about the volume and density of human lung with different races were established for anatomy, anthropology and biodynamics. In two cases of over-load in circulation and water toxicosis, the severe lung edema was proved with the scale of lung density, which was atypical edema in the lung tissue.@*CONCLUSION@#Measurement of visceral organs in the autopsy can assist to an integrative pathological diagnosis.

The study of abnormal glucose and short term prognosis of emergency percutaneous coronary intervention for acute myocardial infarction / 中国基层医药

Objective To investigate the recent prognosis of stress hyperglycemia for the emergency percuta- neous coronary intervention(PCI)with ST-segment elevation acute myocardial infarction(AMI).Methods 410 pa- tients treated by emergency PCI were chosen.According to a history of diabetes and blood glucose levels,they were randomly divided into four groups:group 1(n = 288):random blood glucose normal and non-diabetic patients;group 2(n = 30):random normal blood glucose in diabetic patients;group 3(n = 54):random plasma glucose level and non- diabetic patients;group 4(n = 38):high random plasma glucose level in diabetic patients.Age,gender,hospital within 24 hours with random glucose,glycated hemoglobin(HbA1c),peak creatine kinase,TIMI flow recovery and 30-day mortality of the patients were compared.Results Mortality of four groups were 4.2%,3.4%,7.5% and 5.9% re- spectively(P

Effect of immunocyte therapy on benzene-induced bone marrow haemopoietic dysfunction / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the effect of treatment with immunocyte therapy on benzene-induced haemopoietic dysfunction.</p><p><b>METHODS</b>Mono-nuclear cells (MNC) were separated from 40 - 50 ml peripheral blood in patients and mixed with interleukin-2 and granulocyte macrophage colony stimulating factor (GM-CSF) for six day cultivation. The new formed immunocytes were collected and transfused into the patients. Bone marrow aspiration and biopsy were taken before and after therapy for all patients with severe benzene poisoning. Blood samples were stained by flow cytometry for detecting CD(4) and CD(8) positive cells.</p><p><b>RESULTS</b>Of 20 patients with chronic benzene poisoning, 9 were severe benzene poisoning. All examination including blood count, bone marrow biopsy and T cell subpopulation restored to normal after immunocyte therapy. Laboratory tests (liver and kidney function, and myocardial enzymes) were observed periodically and showed normal during therapy. Follow-up study (the longest time was more than 15 months) showed that bone marrow haemopietic function of all treated patients were in normal range.</p><p><b>CONCLUSION</b>Bone marrow haemopoietic dysfunction caused by benzene poisoning may be closely related to disorder of immune function. Immunocyte therapy may significantly improve bone marrow haemopoietic dysfunction induced by benzene poisoning.</p>