ObjectiveTo investigate the protective effect of erythropoietin (EPO) on human renal tubular epithelial-mesenchymal transition (EMT) induced by high glucose. MethodsCultured HK-2 cells were divided into 5 groups: normal control group, osmolarity control group, high glucose group, high glucose with EPO (5 U/ml)group and high glucose with EPO (10 U/ml)group. The mRNA expression of α-SMA, TGF-β1, Smad2, ILK of cells were measured by RT-PCR. The levels of intracellular α-SMA and TGF-β1 protein were measured by immunocytochemistry. ResultsCompared with the control groups, mRNA expression of α-SMA, TGF-β1, Smad2, ILK and protein expression of α-SMA and TGF-β1 were increased after high glucose treatment (P<0.01). Compared with the high glucose group, EPO 5 U/ml or EPO 10 U/ml remarkably down-regulated the expression of α-SMA, TGF-β1, Smad2 and ILK (P<0.01). ConclusionEPO can inhibit the progression of EMT and the up-regulation of TGF-β1, Smad2 and ILK induced by high glucose.

The aim of this study was to observe the inhibitory effect of daunorubicin on KG1a cells and the expression of Eps8 which is a novel tumor-associated antigen with its full name epidermal growth factor receptor pathway substrate 8 (Eps8), and to explore the effect of daunorubicin on Eps8 expression in KG1a cells at mRNA and protein levels. The KG1a cells were treated with different concentration of daunorubicin for 24, 48, 72 h, then trypan blue staining was used to detect the inhibitory rate of KGla cells, RQ-PCR and Western blot were used to detect Eps8 mRNA and Eps8 protein expression. The results showed that daunorubicin inhibited the proliferation of KG1a cells in a dose and time dependent manner (r = 0.983, P < 0.01). Daunorubicin could reduce the mRNA and protein levels of Eps8 expression in dose and time dependent manners in KG1a cells (r = 0.979, P < 0.05). It is concluded that with the increasing of concentration and time of daunorubicin acting on KG1a cells, the cell proliferative inhibitory effect increased and the expression of Eps8 decreased, suggesting that the inhibitory effect of daunorubicin on KG1a cell proliferation is realized through downregulation of Eps8 expression.
Adaptor Proteins, Signal Transducing ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Daunorubicin ; pharmacology ; Humans

OBJECTIVETo study the expression of neonatal Fc receptor in podocytes in human nephritis and immune-induced rat nephritis models: anti-Thy1.1 nephritis and Heymann nephritis.

METHODSThirty-nine cases of renal biopsies were enrolled from September 2009 to February 2010, including 8 cases of minimal change disease, 4 cases of focal segmental glomerulosclerosis, 9 cases of membranous nephropathy, 12 cases of IgA nephropathy and 6 cases of lupus nephritis. Five normal kidney tissue samples adjacent to renal clear-cell carcinoma were served as normal controls. Laser capture microdissection and real-time RT-PCR were used to assess the expression level of FcRn mRNA in glomeruli of various glomerulonephritides, and immunohistochemistry (IHC) of FcRn by SuperVision method was performed. In addition, rat models of mesangial proliferative nephritis (anti-Thy1.1 nephritis) and passive membranous nephropathy (Heymann nephritis) were established and FcRn was examined in renal tissues by IHC.

RESULTSThe FcRn mRNA level in lupus nephritis was statistically higher than that of normal controls (P < 0.05). FcRn protein expression by IHC was seen in lupus nephritis (6/6), membranous nephropathy (6/9) and IgA nephropathy (7/12), significantly higher than that of normal controls (0/5), P < 0.05. Minimal change disease and focal segmental glomerular sclerosis showed minimal or none expression of FcRn (1/8, 0/4 respectively) and not statistically difference from that of normal controls. Furthermore, FcRn expression in podocytes was detected in rat anti-Thy1.1 (3/5) and Heymann nephritis models (2/7) but was not detected in normal controls.

CONCLUSIONSExpression of FcRn in podocytes was up-regulated in immune-induced human nephritis and rat nephritis models of anti-Thy1.1 nephritis and Heymann nephritis. FcRn may play a role in the development of immune-induced glomerulonephritis.

Animals ; Glomerulonephritis, IGA ; metabolism ; pathology ; Glomerulonephritis, Membranous ; metabolism ; pathology ; Glomerulosclerosis, Focal Segmental ; metabolism ; pathology ; Histocompatibility Antigens Class I ; genetics ; metabolism ; Humans ; Laser Capture Microdissection ; Lupus Nephritis ; metabolism ; pathology ; Male ; Nephritis ; genetics ; immunology ; metabolism ; pathology ; Nephrosis, Lipoid ; metabolism ; pathology ; Podocytes ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Receptors, Fc ; genetics ; metabolism ; Thy-1 Antigens ; immunology ; metabolism ; Up-Regulation

OBJECTIVETo construct COL1A1-targeted short hairpin RNA (shRNA) vector with pSilencer 4.1-CMV neo siRNA expression vector and to evaluate its effect on proliferation and migration of gastric cancer BGC-823 cells in vitro.

METHODSThree COL1A1-shRNA plasmids (COL1A1-shRNA-1, COL1A1-shRNA-2, COL1A1-shRNA-3), targeting different sites of COL1A1 gene, were constructed using pSilencer 4.1-CMV neo siRNA expression vector and transfected into gastric cancer BGC-823 cells. Real time quantitative RT-PCR and Western blot were performed to detect expression levels of COL1A1. MTT and Transwell migration assays were employed to evaluate the effects of COL1A1 gene silence on cell proliferation and migration.

RESULTThree recombinant plasmids targeting COL1A1 were constructed successfully. The expressions of COL1A1 in BGC-823 cells, including mRNA and protein levels, were significantly inhibited by the COL1A1-shRNA transfectants, which resulted in a clear reduction of cell proliferation and migration capacity.

CONCLUSIONThe COL1A1-shRNA can effectively knock down gene expression and inhibit proliferation and migration of gastric cancer BGC-823 cells.

Cell Line, Tumor ; Cell Proliferation ; Collagen Type I ; genetics ; metabolism ; Genetic Vectors ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; pathology ; Transfection ; Transformation, Bacterial

AIMTo develop S-(2-18F-fluoroethyl)-L-methionine (18FEMET) as an amino acid positron emission tomography (PET) tracer for tumors, and to evaluate the value of 18FEMET in the differentiation of experimental tumor and experimental inflammation.

METHODS18FEMET was prepared by nucleophilic fluorination reaction via a two-step procedure. Biodistribution of 18FEMET in normal mice, carcinoma-bearing mice and inflammatory mice, and 18FEMET PET imaging for carcinoma-bearing mice and inflammatory mice were performed compared with 2-[18F] fluoro-2-deoxy-D-glucose (FDG) and O-(2-[18F] fluoroethyl)-L-tyrosine (FET).

RESULTSThe overall radiochemical yield with no decay correction was 15%-25%, the whole synthesis time was about 70 min by manual operation, and the radiochemical purity was above 95%. High uptake and long retention of 18FEMET in pancreas, kidney, colon, liver and heart were observed. But low uptakes in brain and blood were found. Furthermore, high uptake of 18FEMET, FDG and FET in tumor, high uptake of FDG in inflammatory tissue, and almost no uptake of 18FEMET and FET in inflammatory tissue were also observed.

CONCLUSION18FEMET is easy to prepare and can be used to differentiate between tumor and inflammatory tissue. It seems to be a potential amino acid tracer for tumors with PET imaging.

Animals ; Fluorodeoxyglucose F18 ; pharmacokinetics ; Inflammation ; diagnostic imaging ; Methionine ; analogs & derivatives ; chemical synthesis ; pharmacokinetics ; Mice ; Neoplasm Transplantation ; Radiopharmaceuticals ; chemical synthesis ; pharmacokinetics ; Sarcoma 180 ; diagnostic imaging ; pathology ; Tissue Distribution ; Tomography, Emission-Computed ; Tumor Cells, Cultured ; Tyrosine ; analogs & derivatives ; chemical synthesis ; pharmacokinetics

OBJECTIVETo evaluate the diagnostic utility of Warthin-Starry silver stain, immunohistochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease (CSD).

METHODSThe paraffin-embedded lymph node tissues of 77 histologically-defined cases of cat scratch disease collected during the period from January, 1998 to December, 2008 were retrieved and studied using Warthin-Starry silver stain (WS stain) and mouse monoclonal antibody against Bartonella henselae (BhmAB stain). Five cases rich in bacteria were selected for transmission electron microscopy.

RESULTSUnder electron microscope, the organisms Bartonella henselae appeared polymorphic, round, elliptical, short rod or bacilliform shapes, ranged from 0.489 to 1.110 microm by 0.333 to 0.534 microm and often clustered together. Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0% (47/77) of CSD by WS stain. The organisms were located outside the cells and lie mainly in the necrotic debris, especially near the nodal capsule. In 72.7% (56/77) of the cases, dot-like, granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization. Positive results for both stains were identified in 59.7% (46/77) of the cases. When applying both stains together, Bartonella henselae was observed in 74.0% (57/77) of the case. The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance (P < 0.05).

CONCLUSIONSBartonella henselae is the causative pathogen of cat scratch disease. WS stain, BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis. Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Bartonella henselae ; immunology ; isolation & purification ; ultrastructure ; Cat-Scratch Disease ; diagnosis ; microbiology ; pathology ; Child ; Child, Preschool ; Humans ; Immunohistochemistry ; methods ; Infant ; Lymph Nodes ; pathology ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Paraffin Embedding ; Staining and Labeling ; methods ; Young Adult

Objective To study the clinical and laboratory characteristics of adult Japanese encephalitis (JE) patiens in a JE outbreak in Yuncheng, Shanxi province in 2006. Methods All the clinical data from the Second People's Hospital in Yuneheng city were analyzed, part of patients' sera and cerebrospinal fluid were tested by serology and RT-PCR. Results The majority of patients were middle-aged and elderly, 77.8 % (35/45)of the total cases were more than 40 years old. Severe and fulminating type eases accounted for 60.0% (27/45) . Most patients had underlying diseases. IgM antibody to JE virus (JEV) in serum was positive in each of the 45 patients analyzed and 4-fold or greater rise in sera neutralization antibody titer were found in convalescent serum. JEV nucleic acid was positive in part of cerebrospinal fluid specimens. Conclusion Viral encephalitis emerged in Yuncheng city, Shanxi province was Japanese encephalitis B, and most of the eases belonged to elderly group.

Objective To investigate the clinical efficacy of Da Vinci robot-assisted radical resection for right colon cancer.Methods The retrospective cross-sectional study was conducted.The clinical data of 85 patients with right colon cancer who were admitted to the First Hospital Affiliated to Army Medical University from August 2013 to February 2019 were collected.There were 56 males and 29 females,aged from 29 to 84 years,with an average age of 60 years.All patients underwent Da Vinci robot-assisted radical resection of right colon cancer,named right hemicolon D3 + complete mesocolic excision,and received infection prevention and total parenteral nutrition treatment after surgery.According to clinical pathological staging of guideline issued by National Comprehensive Cancer Network,patients underwent postoperative chemotherapy within 1 year after surgery.Observation indicators:(1) treatment status;(2) postoperative pathological examination;(3) follow-up.Follow-up was conducted using outpatient examination,telephone interview and mail every 3 months within 1 year after surgery,every 6 months from 1 to 3 years after surgery,and once a year from 3 to 5 years after surgery up to March 2019.The postoperative tumor metastasis and survival of patients were obtained.Measurement data with normal distribution were represented as Mean±SD,and measurement data with skewed distribution were described as M (range).Count data were expressed as absolute number.Survival rates were calculated using life-table method.Results (1) Treatment status:85 patients underwent Da Vinci robot-assisted right hemicolon D3 + complete mesocolic excision successfully.The operation time,volume of intraoperative blood loss,time for postoperative outof-bed activities,time to recovery of gastrointestinal function,time to liquid diet intake were (178±28) minutes,(85±33) mL,(2.9± 1.8) days,(3.1 ± 2.7) days,(3.9± 1.9) days,respectively.There was no perioperative death.Eleven patients had postoperative complications including 5 of anastomotic leakage,2 of anastomotic bleeding,2 of pulmonary infection,1 of gastric emptying disorder and 1 of incomplete intestinal obstruction;they were cured and discharged after conservative treatment.All the 85 patients received postoperative infection prevention and total parenteral nutrition support,including 64 receiving systemic intravenous chemotherapy with 6 -8 cycles of FOLFOX or XELOX,7 receiving 6-8 cycles of oral capecitabine,and 14 receiving no chemotherapy.(2) Postoperative pathological examination:the number of harvested lymph nodes was 20± 11 and 25 had lymph node metastasis.The length of proximal and distal cutting edge of the specimens was (16±5) cm and (9±5)cm,respectively.There was no cancerous cell on the cutting edge.High-differentiated adenocarcinoma,moderatedifferentiated adenocarcinoma,moderate-differentiated tubular adenocarcinoma,low-differentiated adenocarcinoma,mucinous adenocarcinoma,tubular combined with mucinous adenocarcinoma were detected in 2,40,14,16,9,4 patients,respectively.There were 8,28,24,5,12,8 patients in Ⅰ stage,Ⅱ A stage,Ⅱ B stage,Ⅱ C stage,ⅢB stage,Ⅲ C stage of TNM staging,respectively.(3) Follow-up:85 patients were followed up for 1-67 months,with a median follow-up time of 19 months.During the follow-up,1 of 85 patients had liver metastasis at 14 months after surgery and had survived after radiofrequency ablation treatment up to the end of follow-up.Three cases died of abdominal tumor metastases,1 of which in Ⅱ C stage died at 32 months after surgery,1 in Ⅲ B stage died at 4 months after surgery and 1 in Ⅲ B stage died at 16 months after surgery.The 1-,3-year overall survival rates were 97.1％ and 94.0％,respectively.Conclusion Da Vinci robot-assisted radical resection of right colon cancer is safe and feasible,with good short-and long-term outcomes.

Objective:To investigate the availability, prices and affordability of essential medicines in pediatric population across China, in the hope of improving rational use of medicines.Methods:A multicenter cross-sectional survey of medicine prices, availability and affordability was conducted in 17 provinces, municipalities and autonomous region across east, south-central part, west and north of China. Data on 42 medicines used in pediatric population, both original and generic, were collected in 55 public hospitals from May 26 to June 2, 2017. Availability was expressed as the percentage of hospitals with stock of the target medicine on the day of data collection,and median price ratio (MPR) was the ratio of price upon investigation to international reference. Based on national minimum daily wage, affordability represents the number of working days needed to earn the expense which covers a standard course using the target medicine. Statistical software SPSS 13.0 was applied for descriptive analysis of availability, MPR and affordability.Results:Mean Availability of original and generic medicine was 33% and 32%, with median MPR being 5.43 and 1.55. Among the 19 medicines with price information for both original and generic product, the median MPR was 7.73 and 2.04 respectively. Regarding the five medicines used to treat four common pediatric diseases (pneumonia,peptic ulcer, congenital hypothyroidism, refractory nephrotic syndrome), the affordability was 0.63 (0.16-6.17) d for generic medicine, and 1.03 (0.16-11.53) d for its original counterpart.Conclusions:The availability to both original and generic products of the 42 medicines used in pediatric population was low in China. The prices of generic medicines seem to be lower and affordability higher than those of original medicines. There is an urgent need to improve the availability and affordability of pediatric medicines.

Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.