OBJECTIVETo investigate the mechanism of beta-cell dysfunction induced by glucolipotoxicity in high fat-fed obese rats.

METHODSEighteen high-fat obese male Wistar rats were assigned into 3 groups and underwent 48-hour infusion through the jugular vein with normal saline (n=6), 20% intralipid + heparin (FFA group, n=6), or 25%glucose +20% intralipid + heparin (GS-FFA group, n=6). The plasma beta-hydroxybutyric acid (beta-HBA) was measured before and at the end of the infusion. After the infusion, the rats were sacrificed following an intravenous glucose tolerance test (IVGTT) to remove the tail of the pancreas for detection of apoptotic islet cells using TUNEL method. Immunohistochemical staining was performed to detect the expression of cytochrome c (cyt c), apoptosis-inducing factor (AIF), caspase-9 and caspase-3 in the islet cells.

RESULTSAt the end of the infusion, all the rats exhibited increased plasma beta-HBA levels, which was the highest in the GS-FFA group (P<0.05). IVGTT performed after the infusion showed a significantly lower insulinogenic index in GS-FFA group than that in NS and FFA groups. Greater number of apoptotic islet cells was found in the GS-FFA group than in the FFA and NS groups (P<0.05), and the islets had significantly higher levels of cyt c, AIF, caspase-9 and caspase-3 in the former group than in the latter two groups (P<0.05).

CONCLUSIONSHyperglycemia and high free fatty acid level synergistically impair insulin secretions to cause ketone overproduction in high fat-fed obese rats. The beta-cell dysfunction due to glucolipotoxicity is associated with increased beta-cell apoptosis and activation of mitochondrial apoptotic pathway.

3-Hydroxybutyric Acid ; blood ; Animals ; Apoptosis ; drug effects ; Fat Emulsions, Intravenous ; pharmacology ; Glucose ; pharmacology ; Glucose Tolerance Test ; Insulin-Secreting Cells ; cytology ; pathology ; Male ; Mitochondria ; drug effects ; Obesity ; physiopathology ; Rats ; Rats, Wistar

OBJECTIVETo obtain isolated human metapneumovirus (HMPV) strains from clinical specimens collected from infants and children in Beijing and to promote the investigation on this important respiratory pathogen.

METHODClinical specimens including throat swabs from outpatients and nasopharyngeal aspirates from hospitalized children were collected from infants and children visited the affiliated children's hospital for acute respiratory infections during May 2008 to April 2009. HMPV positive specimens identified by RT-PCR and/or direct immunofluorescent assay with monoclonal antibody against HMPV were inoculated to LLC-MK(2) cells and incubated at 37°C and 33°C, respectively. The replication of the virus in the cells was detected by direct immunofluorescent assay followed by RT-PCR. The genotypes of the isolated virus strains were identified by RT-PCR.

RESULTOut of 1092 clinical specimens, 81 were HMPV positive by RT-PCR, the positive rate was 7.4% (81/1092). Among these positive specimens, 33 were inoculated to LLC-MK(2) cells and the replication of HMPV was revealed by antigen detection and RT-PCR from 5 out of these 33 inoculates. These isolated viruses could be passed in LLC-MK(2) cells and were not cross-reacted with other common respiratory viruses, such as ADV, RSV and Parainfluenza viruses 1/2/3 by monoclonal antibodies against these viruses in direct immunofluorescent assay. The HMPV was more likely to be isolated from fresh specimens within 24 hours after the collection of specimens which were not frozen. Four of the 5 isolated strains were identified as genotype A and 1 as genotype B. Unlike other respiratory viruses, these isolated HMPV did not show specific CPE in cell culture and the replication of the virus was identified by antigen detection and RT-PCR.

CONCLUSIONHMPV of both genotypes were isolated from infants and children with acute respiratory infections in Beijing which will accelerate the investigation of this important virus.

Acute Disease ; Child ; China ; Genes, Viral ; genetics ; Genotype ; Humans ; Infant ; Metapneumovirus ; isolation & purification ; Respiratory Tract Infections ; virology

OBJECTIVESVentricular remodeling is an important pathologic progress in almost all end stage heart failure (HF), and it is characterized by ventricular thickening and cardiac fibrosis with poor prognosis. The connective tissue growth factor (CTGF), a new growth factor with multi-function, has an important role in fibrosis of tissue and organs. It has been demonstrated that angiotensin-converting enzyme inhibitor (ACEI) can prevent the development of cardiomyocyte from remodeling and improve cardiac function. Researchers try to test the hypothesis that cardiac function improvement attributable to ACEI is associated with inhibiting expression of CTGF in patients with HF. The aim of this study was to observe changes in CTGF expression in cardiomyocyte of young rats with HF and effect of benazepril on CTGF.

METHODSThe animal model of HF was established by constriction of abdominal aorta. Five weeks old rats were randomly divided into 3 groups after 6 weeks of operation: (1) HF group without treatment (n = 15); (2) HF group where rats were treated with benazepril (n = 15); (3) sham-operated group (n = 15) where rats were administered benazepril through direct gastric gavage. After 4 weeks of treatment, the high frequency ultrasound was performed. The expression of CTGF was detected by immunohistochemistry and semi-quantative reverse transcription-polymerase chain reaction.

RESULTSCompared with the sham-operated group, left ventricular diastolic dimension (LVEDD), left ventricular systolic dimension (LVESD), interventricular septal thickness at end-diastole (IVSTd), interventricular septal thickness at end-systole (IVSTs), left ventricular posterior wall thickness at end-diastole (LVPWTd), left ventricular posterior wall thickness at end-systole (LVPWTs), left ventricular relative weight (LVRW), and right ventricular relative weight (RVRW) were all increased (P < 0.01), but ejection fraction (EF) and fractional shortening (FS) were decreased (P < 0.01). CTGF positive cells and expression of CTGF mRNA (0.609 +/- 0.065 vs 0.117 +/- 0.011, P < 0.01) were increased in HF group without treatment. LVESD, IVSTd, IVSTs, LVPWTd, LVPWTs, LVRW and RVRW were all decreased (P < 0.01), but FS and EF were increased (P < 0.01) in cases of HF treated with benazepril when compared with HF group without treatment. LVESD, IVSTd, IVSTs, LVPWTd, LVPWTs, LVRW and RVRW were higher (P < 0.01), EF and FS were lower (P < 0.01), CTGF positive cells and expression of CTGF mRNA were higher (P < 0.01) in HF group treated with benazepril than those of sham-operated group.

CONCLUSIONThe expression of CTGF was increased in the cardiomyocyte of young rats with HF and benazepril could prevent left ventricular from remodeling partly and improve cardiac function by inhibiting the expression of CTGF in cardiomyocyte in cases of HF.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Benzazepines ; pharmacology ; Connective Tissue Growth Factor ; metabolism ; Disease Models, Animal ; Heart Failure ; diagnostic imaging ; drug therapy ; metabolism ; physiopathology ; Immunohistochemistry ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; RNA, Messenger ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Ultrasonography ; Ventricular Dysfunction, Left ; diagnostic imaging ; drug therapy ; physiopathology ; Ventricular Remodeling ; drug effects

BACKGROUNDDense congenital cataracts often cause severe visual impairment. The results of long-term follow-up of dense bilateral congenital cataract in China have not been well documented. The purpose of this study was to evaluate the long-term visual function in children who underwent cataract extraction for dense bilateral congenital cataract in southern part of China.

METHODSMedical records of children who underwent surgery of dense bilateral congenital cataract between January 1992 and December 2000 at Zhongshan Ophthalmic Center of Sun Yat-sen University were retroactively reviewed. In 38 children available for current follow-up, best corrected visual acuity (BCVA) and stereoscopic vision, as well as nystagmus, strabismus, and other complications, were evaluated. The mean follow-up period was 107.6 months (range 60 to 167 months).

RESULTSThe mean age of cataract extraction and secondary intraocular lens implantation were 5.6 months (range 3 to 12 months) and 4.2 years (range 2.4 to 15 years), respectively. The mean BCVA was 0.25 in the better eye and 0.16 in the fellow eye. Stereoscopic vision was absent in all patients, and 3 children had simultaneous perception. Nystagmus was detected in all cases and strabismus in 35 cases. A high correlation was found between timing of cataract extraction and final BCVA of the better eye (r = -0.55, P = 0.00). A statistically significant difference was found in BCVA between post- and pre-treatment of amblyopia (t = 5.65, P = 0.00).

CONCLUSIONSLong-term visual function in children with dense bilateral congenital cataract was poor when cataract surgery was performed at age of 3 months or later. Earlier cataract surgery with adequate optical rehabilitation contributed to better visual outcome.

Adolescent ; Cataract ; congenital ; Cataract Extraction ; Child ; Child, Preschool ; Follow-Up Studies ; Humans ; Infant ; Retrospective Studies ; Visual Acuity

Objective To investigate the relationship between perioperative nutritional risk and venous thromboembolism(VTE)in patients with hip fracture.Methods A total of 379 patients with unilateral hip fracture due to fall or sprain who underwent elective surgery were selected and divided into the non-VTE group(246 cases)and the VTE group(133 cases)according to whether or not VTE occurred during perioperative period.Basic information,surgical and anesthesia records,nutritional risk related indicators,inflammatory indicators and outcome indicators of patients were collected.Multiple Logistic regression was used to analyze the independent influencing factors of perioperative VTE.Receiver operating characteristics(ROC)curves were used to assess the ability to discriminate independent factors,and DeLong test was used to compare area under the curve(AUC).Results Compared with the non-VTE group,the proportion of patients in the VTE group was older,complicated with hypertension,the time to visit hospital more than 2 days,received(hollow/intramedullary nail)internal fixation,perioperative blood transfusion,ASA gradeⅢtoⅣ,and higher nutritional risk screening Table(NRS)-2002 scores on admission and higher postoperative neutrophil/lymphocyte ratio(NLR).Nutritional prognosis index(PNI),hemoglobin(Hb)and prealbumin(PA)at admission and after operation were lower in the VTE group than those in the non-VTE group(P＜0.01).Multivariate Logistic regression analysis showed that PNI was decreased,NRS-2002 scores and PA were increased,and the time of visit hospital was>2 days after internal fixation.American College of Anesthesiologists(ASA)gradesⅢ-Ⅳwere independent risk factors for perioperative VTE of hip fracture(P＜0.05).ROC curve analysis showed that the AUC(95%CI)of NRS-2002 at admission was 0.739(0.692-0.783),and that of PNI at admission was 0.720(0.672-0.765),both of which were better than other influencing factors(P＜0.01).Conclusion NRS-2002 and PNI are good predictors of perioperative VTE in patients with hip fracture.

Background:Accumulating studies on computational fluid dynamics (CFD) support the involvement of hemodynamic factors in artery stenosis.Based on a patient-specific CFD model,the present study aimed to investigate the hemodynamic characteristics of transplant renal artery stenosis (TRAS) and its alteration after stent treatment.Methods:Computed tomography angiography (CTA) data of kidney transplant recipients in a single transplant center from April 2013 to November 2014 were reviewed.The three-dimensional geometry of transplant renal artery (TRA) was reconstructed from the qualified CTA images and categorized into three groups:the normal,stenotic,and stented groups.Hemodynamic parameters including pressure distribution,velocity,wall shear stress (WSS),and mass flow rate (MFR) were extracted.The data of hemodynamic parameters were expressed as median (interquartile range),and Mann-Whitney U-test was used for analysis.Results:Totally,6 normal,12 stenotic,and 6 stented TRAs were included in the analysis.TRAS presented nonuniform pressure distribution,adverse pressure gradient across stenosis throat,flow vortex,and a separation zone at downstream stenosis.Stenotic arteries had higher maximal velocity and maximal WSS (2.94 [2.14,3.30] vs.1.06 [0.89,1.15] m/s,256.5 [149.8,349.4] vs.41.7 [37.8,45.3] Pa at end diastole,P =0.001;3.25 [2.67,3.56] vs.1.65 [1.18,1.72] m/s,281.3 [184.3,3 64.7] vs.65.8 [61.2,71.9] Pa at peak systole,P =0.001) and lower minimal WSS and MFRs (0.07 [0.03,0.13] vs.0.52 [0.45,0.67] Pa,1.5 [1.0,3.0] vs.11.0 [8.0,11.3] g/s at end diastole,P =0.001;0.08 [0.03,0.19] vs.0.70 [0.60,0.81] Pa,2.0 [1.3,3.3] vs.16.5 [13.0,20.3] g/s at peak systole,P =0.001) as compared to normal arteries.Stent implantation ameliorated all the alterations of the above hemodynamic factors except low WSS.Conclusions:Hemodynamic factors were significantly changed in severe TRAS.Stent implantation can restore or ameliorate deleterious change of hemodynamic factors except low WSS at stent regions.

Sarcopenia,osteoporosis and osteosarcoporosis,which belong to the same category of musculoskeletal unit disease,seriously impair the homeostasis of the musculoskeletal system in the aging process of the human body.In recent years,researchers have applied various emerging technologies to conduct multi-disciplinary,multi-level and multi-dimensional research on musculoskeletal decay disease,and have come to realized that the disease involves complex biochemical and mechanical crosstalk,and the intestinal microecology has also aroused great interest in this crosstalk because of its profound impact on musculoskeletal health.In view of this,this paper takes the close relationship between intestinal microecology and musculoskeletal decay disease as the starting point,and under the guidance of the overall concept,we deeply analyzes the mechanism and regulatory information behind this relationship,and on this basis,we excavates the potential therapeutic targets for musculoskeletal decay disease,so that we can provide new ideas and strategies for the management of the disease in the context of the musculoskeletal subunit.

BACKGROUNDHuman metapneumovirus (hMPV) was discovered by scientists in the Netherlands as a novel respiratory virus in 2001 and had been found in children with acute respiratory tract infections (ARTI) in China. The objective of this study was to determine the importance of hMPV infection in children in Beijing and the genotypes of the circulating virus by the surveillance during a four-consecutive-year period.

METHODSClinical specimens collected from children with ARTI from January 2006 to December 2009 were tested for hMPV by RT-PCR using primers targeting the matrix (M) gene, followed by genotyping of hMPV directly from positive samples by diplex PCR with primers for glycoprotein (G) genes. Sequence analysis was used for genotyping of those un-typable samples. Common respiratory viruses in these clinical specimens were tested by virus isolation and antigen detection, in addition to hMPV detection.

RESULTSOf 4730 tested specimens, 191 (4.0%) were positive for hMPV and 62.8% of 191 were identified as genotype A. The positive rate of hMPV from hospitalized patients was higher than that from outpatients each year. Most of hMPV positive children were under five years old. The peak of hMPV activity mostly occurred in late spring and overlapped with or followed that of respiratory syncytial virus (RSV) and followed by parainfluenza virus 3. Of hMPV infected cases, 68.6% were lower respiratory tract infection, among which 79.4% were hospitalized, and upper respiratory tract infection was diagnosed for 31.4% of hMPV infected children. The 9.4% of hMPV positive samples were found to co-exist with other respiratory viruses.

CONCLUSIONShMPV was an important pathogen for ARTI in pediatric patients, especially those under five years old. Both genotypes A and B circulated simultaneously in Beijing.

Adolescent ; China ; Female ; Genotype ; Humans ; Male ; Metapneumovirus ; genetics ; pathogenicity ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEThe aim of the study was to evaluate the anti-HBs persistence and the long term preventive efficacy after vaccination 23 years with plasma-derived hepatitis B vaccine.

METHODSThe study consisted of 261 children who were 5 - 9 years aged, from two primary schools in two townships of Xi'an. 126 children were randomly selected as vaccine group, and 135 children in control group. These children were followed up again in 2009. Excluding self-inoculation, the vaccine and control groups were 81 and 75, who was used to ask to recall details of their experience for vaccination and liver-related illnesses during past twelve years. Individuals who had anti-HBs titers less 10 mIU/ml, HBsAg, anti-HBc and HBV-DNA all were negative, were given a booster dose vaccine and retest for anti-HBs titer after one month.

RESULTSAfter eliminated the interference of an early booster dose and vaccination outside the study, the positive rate of anti-HBs was 48.1% (39/81) in the vaccine group at year 23, higher than 34.7% (26/75) in control group. At year 23 after primary vaccination, 84.0% (21/25) individuals in the vaccine group whose anti-HBs and anti-HBc both are negative showed a stronger anamnestic response after received a booster dose, while 7.5% (3/40) in the control group. At year 23 after primary vaccination, none clinical case of hepatitis B was found among 194 individuals. However, anti-HBc positive rate in the vaccine group was 16.0% (13/81), while the rate in the control group was 30.7% (23/75) (χ(2) = 4.687, P < 0.05).

CONCLUSIONAt 23 years after implemented a full course of plasma-derived hepatitis B vaccine, the recipients of vaccine were maintained anti-HBs at a high level or strong immunological memory.

Child ; Child, Preschool ; Follow-Up Studies ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; Humans ; Immunization, Secondary ; Immunologic Memory ; immunology ; Plasma ; immunology

OBJECTIVETo identify variations in hemagglutinin genes from influenza viruses (H3N2) isolated from infants and young children with acute respiratory infection (ARI) between March, 2004 and April 2005.

METHODSRNAs from influenza A virus strains (subtype H3) isolated from specimens collected from ARI children were extracted followed by amplification for HA1 fragments from hemagglutinin (HA) genes by RT-PCR. The sequences of the fragments were defined by direct sequencing for the PCR products or the target inserts after the PCR fragments were cloned into the TA-cloning vector pBS-T and analyzed by bioinformatic software.

RESULTSFragments of 987 bps of HA1 (encoding 329 amino acids) from a total of 32 strains of influenza A virus (subtype H3) isolated from the 2004 season and 1 from the 2003 season were amplified and the sequences were compared with vaccine reference strains recommended by WHO which were used in recent years. There were several consistent amino acid variations which involved in both antigenic epitopes A and B and receptor binding site (RBS) for isolated strains in the 2004 influenza season compared with the vaccine strains used during the recent years and the virus strains isolated in March 2004, indicated the antigenic drift of the viruses isolated in 2004 influenza season may lead to variant viruses.

CONCLUSIONThe variations of the HA genes from influenza virus (subtype H3) strains in the 2004-2005 influenza season were confirmed by sequence analysis for the HA1 regions of the hemagglutinin genes, which indicate that the antigenic drift would have been caused by the diversification of the genes and the efficacy of the recently used vaccines should be kept under close watch.

Antigenic Variation ; Child ; China ; epidemiology ; Evolution, Molecular ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; immunology ; virology ; RNA, Viral ; genetics ; Sequence Analysis, RNA