Detection and analysis of children with severe community-acquired pneumonia using automatic nested multiplex PCR system / 中国小儿急救医学

Objective:To investigate the effect of automated nested multiplex PCR system in the detection of children with severe community-acquired pneumonia(CAP), and identify the pathogenic infection of the children with severe CAP in Foshan.Methods:Children with severe CAP, who were admitted to the PICU at Foshan First People′s Hospital from January 2016 to December 2018, were enrolled in the analysis.Nasopharyngeal secretions were collected.And automated nested multiplex PCR was used to detect adenovirus, coronavirus (HKUl type, NL63 type, 229E type, 0C43 type), human metapneumovirus, influenza A virus (H1 subtype, H1-2009 subtype, H3 subtype), influenza B virus, parainfluenza virus (type 1, type 2, type 3, type 4), respiratory syncytial virus, Bacillus pertussis, Chlamydia pneumoniae and Mycoplasma pneumonia.Results:Among the 290 specimens detected by the automated nested multiplex PCR, 246(84.83%) were positive.There were 166 positive samples for a single pathogen, 60 positive samples for two pathogens, 17 positive samples for three pathogens, and three positive samples for four pathogens.Among the virus-positive cases, respiratory syncytial virus was the most common pathogen in children younger than 6 months(62.39%, 73/117). The most common pathogen was human rhinovirus/enterovirus(43.48%, 20/46) from seven months to one year old.Adenovirus(37.50%, 18/48) was the most common pathogen among children aged one to three years old.Rhinovirus/enterovirus(35.00%, 7/20) was the most common pathogen among children aged three to six years old.The most common pathogen in children over six years old was influenza virus(46.67%, 7/15). The adenovirus detection rate was highest in May, the syncytial virus detection rate was highest in August, and the influenza virus detection rate was highest in July.Mycoplasma pneumoniae and pertussis were distributed throughout the year.Conclusion:The automated nested multiplex PCR system can detect multiple pathogens efficiently, quickly and accurately; the common pathogens of severe CAP are diverse in different age groups; the epidemic season of common pathogens is unique in different regions due to different climates.

Molecular cloning and spatiotemporal expression of an APETALA1/FRUITFULL-like MADS-box gene from the orchid (Cymbidium faberi) / 生物工程学报

In order to identify genes involved in floral transition and development of the orchid species, a full-length APETALA1/FRUITFULL-like (AP1/FUL-like) MADS box cDNA was cloned from Cymbidium faberi (C. faberi) sepals and designated as C. faberi APETALA1-like (CfAP11], JQ031272.1). The deduced amino acid sequence of CfAP11 shared 84% homology with a member of the AP1/FUL-like group of MADS box genes (AY927238.1, Dendrobium thyrsiflorum FUL-like MADS box protein 3 mRNA). Phylogenetic analysis shows that CfAP11 belonged to the AP1/FUL transcription factor subfamily. Bioinformatics analysis shows that the deduced protein had a MADS domain and a relatively conservative K region. The secondary structure of CfAP11 mainly consisted of alpha helices (58.97%), and the three-dimensional structure of the protein was similar to that of homologues in Roza hybrida, Oryza sativa and Narcissus tazetta. Real-time quantitative PCR (qRT-PCR) results reveal low levels of its mRNA in roots, lower levels in leaves during reproductive period than vegetative period, and higher levels in pedicels at full-blossom stage than at bud stage. These results suggest that CfAP11 is involved in floral induction and floral development. Additionally, we observed higher levels of CfAP11 expression in pedicels and ovaries than in other tissues during full-blossom stage, which suggests that CfAP11 may also be involved in fruit formation in certain mechanism.

The prevalence and risk factors of kidney disease in type 2 diabetic patients in rural Shanghai / 中华内科杂志

Objective To identify the prevalence and etiology of kidney disease and the related risk factors in type 2 diabetic patients in rural Shanghai.Methods A cross-sectional study in type 2 diabetic patients was conducted in a community of Shanghai.Questionnaire,clinical examination and laboratory tests were completed to collect the information about sociodemographic and healthcare characteristics.Results A total of 1421 eligible patients with complete information were screened from 1487 type 2 diabetic patients between November 2008 and March 2009.Of them,40.75％ were men,59.25％ were women,aged 37-86 (61.33 ± 9.65 ) years old,with diabetic duration of 0.25-43.92 (7.85 ± 6.34) years.Among them,43.42％ had diabetic retinopathy,21.18％ had neuropathy; 69.95％ met the screening definition for hypertension,76.07％ for hyperlipidemia,15.55％ for hyperuricemia and 23.65％ for cardiovascular disease.The control rates of fasting blood glucose,glycosylated hemoglobin,blood pressure and serum cholesterol were 57.71％,33.99％,14.22％ and 2.46％,respectively.The prevalence of kidney disease,diabetic nephropathy and non-diabetic renal disease was 41.31％,18.51％ and 13.44％,respectively; and 9.36％ were diagnosed as renal insufficiency of unknown reasons.Age,diabetic duration,hyperuricemia,diabetic retinopathy and poor control of blood pressure were independently associated with kidney disease;age and poor control of blood pressure were independently associated with diabetic nephropathy; age and hyperuricemia were independent risk factors of renal insufficiency in patients with diabetic nephropathy.Conclusions Although the diabetic duration of these subjects is relatively short,the prevalence of complications including diabetic nephropathy is high.The high prevalence of non-diabetic renal disease shows the importance of further screening and diagnoses for prevention.Strict control of blood glucose,blood pressure,serum cholesterol and serum uric acid are key points of cutting down the prevalence of diabetic nephropathy and chronic kidney disease.

Clinicopathology, diagnosis and classification of renal amyloidosis / 中华肾脏病杂志

ObjectiveTo clarify the clinicopathological features of renal amyloidosis in order to achieve early diagnosis and treatment.MethodsClinicopathological data of 26 biopsyproven renal amyloidosis cases in Department of Nephrology,Zhongshan Hospital,Fudan University between2006and2010wereanalyzedretrospectively.Immunohistochemistryand immunofluorescence of amyloid A protein,immunoglobulin light chains such as K、λ were performed on renal specimens for further classification.ResultsAge of 26 patients ranged from 40 to 77 years old,average(58.54±10.07) years.Twenty-two out of 26 patients(84.62％) were treated in local hospital before admitted to our department,and 21 patients(95.45％) were misdiagnosed as chronic primary glomerulonephritis.The prominent clinical manifestations of renal amyloidosis were nephrotic syndrome(17 cases,65.38％),decreased blood pressure(16 cases,61.53％),organ enlargement (8 cases,30.77％) and bodyweight loss (6 cases,23.08％).Fourteen out of 25 patients (56.00％) were found to have monoclonal light chains in serum by immunofixation electrophoresis.Three patients with mild pathological changes who had no confirmable Congo red stain were conffimed by electron microscopy. Twenty-three(88.46％) patients werediagnosed as AL amyloidosis,one(3.85％) as AA amyloidosis,one was strongly suspected of hereditary amyloidosis,and one was undetermined.ConclusionsRenal amyloidosis is frequently misdiagnosed.Middleaged and old nephrotic patients with decreased blood presure,organ enlargement and bodyweight loss may be the most helpful clues of the disease.Most patients have monoclonal light chains in serum or urine.Renal biopsy,especially electronic microscopy plays a crucial role in the early diagnosis of renal amyloidosis.Immunohistochemistry is important for patients with renal amyloidosis in pathological classification and treatment.

Tilting the balance of tubular cell fate toward survival contributes to ischemic tolerance in kidney / 中华肾脏病杂志

Objective To explore the role of brief ischemia pretreatment in the induction of renal ischemic tolerance,and investigate its effects on tubular cell necrosis,apoptosis and proliferation. Methods Male Sprague-Dawley rats were randomly divided into three groups,including sham-operated group (Sham),ischemia/reperfusion injured group subjected to theocclusion of both renal pedicles for 40 min followed by reperfusion(I/R),and preconditioned group with 20-min ischemia pretreatment induced 4 days before I/R(IPC).Histological changes were evaluated by PAS staining.The ultra-structure of tubular cells was observed by transmission electron microscopy(TEM).Apoptosis was confirmed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL).The proliferation of tubular cells was evaluated with proliferating cell nuclear antigen(PCNA). Results Twenty-minites ischemia pretreatment offered both promising functional and histological protection against 40-min ischemia/reperfusion injury (P<0.01).The mortality rate wag reduced from 33%in I/R group to 0 in IPC group.The renopmtection offered by 20-min ischemia pretreatment was accompanied with reduced postischemic tubular cell apoptosis and necrosis (P<0.05), and increased cell proliferation (PCNA positive) (P< 0.01). Conclusions Brief and sublethal prior ischemia can render the kidney more tolerant to subsequent prolonged I/R injury. Its ability to tilt the balance of tubular cell fate toward survival, reducing postischemic cell death and enhancing cell proliferation, may play an important role in renal protection of ischemic preconditioning.

Expression and location of hypoxia inducible factor-1α and -2α in the remnant kidney of 5/6 nephrectomy rats / 中华肾脏病杂志

Objective To investigate the location and expression of hypoxia inducible factor (HIF) subunits in the remnant kidney of 5/6 nephrectomy rats. Methods Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at week 1, 2, 4, 6, 8 and 12 after operation. Tissues of remnant kidneys were collected to detect the location and expression of HIF-1α and HIF-2α by immunohistochemistry staining and Western blotting. The mRNA levels of HIF targeted genes vascular endothelial growth factor (VEGF) and heme oxygenase-1 (HO-1) were determined by RTPCR. Results (1) 5/6 nephrectomy rats underwent one week of acute renal failure at first[Scr (122.8±22.1) μmol/L] and then developed compensative chronic renal failure [(66.0±3.7)-(66.4±8.4) μmol/L], but the level of Scr increased quickly after week 6 [(66.4±8.4)-(127.8±22.7) μmol/L],concomitantly with progressive tubulointerstitial fibrosis in remnant kidney cortex. (2) In cortex, HIF-1α was expressed only in the atrophic and dilated tubular cells while HIF-2α was located in endothelial, interstitial fibroblasts, and vascular smooth muscle cells. The semiquantitative results of imunohistochemistry and Western blotting revealed that HIF-1α and HIF-2α were both gradually up-regulated during the early stage of remnant kidney, peaked at week 4 and 6, and then gradually down-regulated. (3) The mRNA levels of HIF targeted genes VEGF and HO-1 transiently peeked at week 4 and 6, and then decreased gradually. Conclusions The increased stabilization of HIF-αprotein and transcription of HIF targeted genes at the early stage of this model is a compensation reaction towards hypoxia. The mechanism of decreased expression of HIF-α at the end stage of chronic kidney disease deserves further investigation.

Hypoxia inducible factor-1α gene silencing aggravates growth inhibition and necrosis of human proximal renal tubular epithelial cell under hypoxia / 中华肾脏病杂志

Objective To explore the effect of hypoxia inducible factor-1α silencing by siRNA on proliferation, apoptosis and necrosis of human renal proximal epithelial cell ( HK-2 )under hypoxia. Methods CoCl2 was used to mimic hypoxia, and siRNA technique was used to silence HIF-1α. HK-2 cells were divided into five groups, including normal culture group,hypoxia culture group, transfection reagent group, negative control group and HIF-1α siRNA group.MTT assay was used to detect the growth inhibition ratio of cells. TUNEL and caspase-3 quantity was used to detect apoptosis. LDH activity in supernatant was detected to evaluate cell necrosis.Real-time PCR and Western blotting were used to detect mRNA and protein of HIF-1α, HIF-2α,glucose transporter 1(Glut-1) and vascular endothelial growth factor (VEGF). Results (1) The transfection efficiency of siRNA was 95%-100%. Under normoxia, the efficiency of siRNA silencing HIF-1α gene reached to 70%, while under hypoxia, it was 97%. (2) The growth inhibition ratio of cells in HIF-1α siRNA group was significantly higher than that of other groups including hypoxia culture group, transfection reagent group and negative control group (all P＜0.05). No significant difference was found in apoptotic ratio of the other four groups except normal culture group (P＞0.05). The LDH level in HIF-1α siRNA group was significantly higher than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). (3) The expression of HIF1α, Glut-1, VEGF mRNA and protein in HIF-1α siRNA group was significantly lower than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). No significant difference was observed on the level of HIF-2α mRNA and protein in the other four groups except normal culture group (P＞0.05). Conclusion HIF-1α gene silencing can inhibit the mRNA and protein expression of Glut-1 and VEGF, and can aggravate growth inhibition and necrosis of HK-2 cells under hypoxia.

Clinicopathological analysis of IgA nephropathy patients with mild proteinuria and/or hematuria / 中华肾脏病杂志

Objective To clarify the relationship between clinical manifestation and pathological features of IgA nephropathy (IgAN) patients with mild proteinuria and/or hematuria.Methods Clinicopathological data from 316 biopsy-proven IgAN cases (proteinuria＜1 g/24 h and/or hematuria, and Scr＜133 μmol/L) from our hospital between January 1993 and October 2009 were studied retrospectively. The renal histopathology was quantified according to Lee's grading and Katafuchi's semi-quantitative standard, and the risk factors for renal pathological lesions were evaluated using multifactor logistic regression analysis. Results Among these 316 patients, 123 were male and 193 patients were female. The mean age at the time of renal biopsy was (33.10±10.69) years old. Clinical features were found as follows: hematuria with proteinuria was found in 267 patients (84.5%), isolated hematuria in 24 patients (7.6%), and isolated proteinuria in 25 patients (7.9%). 16.5% of patients had hypertension. The percentages of CKD stage Ⅰ, Ⅱ, Ⅲ were 76.9%, 20.9% and 2.2%, respectively. 31.3% of patients presented Lee's grade Ⅲ or more severe.52.8% of patients had various degrees of glomerulosclerosis. Crescent formation was observed in 20.3% of patients. 22.5% of patients showed tubular atrophy;16.8% showed interstitial fibrosis and 24.7% also had renal vascular lesions. The extent of glomerulosclerosis was negatively correlated with eGFR levels, but positively correlated with the amount of proteinuria and mean arterial pressure (MAP) level (P＜0.05). The score of tubulointerstitial lesion was positively correlated with the amount of proteinuria and negatively correlated with eGFR and hemoglobin (Hb)level (P＜0.05). The degree of renal vascular lesion was also correlated to MAP level positively and eGFR level negatively (P＜0.05). Multifactor logistic regression analysis revealed that proteinuria, Scr and Hb at the time of renal biopsy were independent risk factors for severe renal pathological lesions (Lee's grade Ⅲ or more severe) with odds ratio of 8.564, 1.031 and 0.975 respectively (all P＜0.01). Conclusions Severe renal histological lesions and decrease of renal function may be seen in some IgAN patients with mild proteinuria and/or hematuria. The levels of proteinuria,Scr and Hb are the independent risk factors for severe renal pathological lesions. Renal biopsy is important in these patients in order to make diagnosis and individual treatment.

A study of therapeutic strategies for idiopathic membranous nephropathy / 复旦学报（医学版）

Objective To compare the efficacy of different therapies for idiopathic membranous nephropathy (IMN) patients, and to discuss their rationality. Methods The clinicopathological data and therapies of 76 patients with IMN in our hospital was retrospectively analyzed and reviewed, and the efficacy was followed up.According to the different therapies, 76 patients were divided into 4 groups, including symptomatic treatment group, glucocorticoid-alone group, immunosuppressant-alone group, and glucocorticoid in combination with immunosuppressant group (combination group). Comparison and analysis of the efficacy of the different therapies were made. Results (1) The incidence of nephrotic syndrome and 24-hour proteinuria of patients in symptomatic treatment group were significantly lower than those in glucocorticoid-alone group and combination group. (2) The remission rates of 4 groups were 56.3%,73.7%,66.7% and 78.9%, respectively. In general, no statistical differences were observed in the remission rates of patients among the symptomatic treatment group, glucocorticoid-alone group and combination group. The 2-year and 5-year renal survival rates were 89.2% and 79.3%, respectively. (3) Patients in glucocorticoid-alone group and combination group were divided into low-risk, moderate-risk and high-risk patients. No difference in remission rate was observed between the two therapies for low-risk and moderate-risk patients.But for high-risk patients,the remission rate in combination group was significantly higher than that in glucocorticoid-alone group.(4) Patients in glucocorticoid-alone group and combination group were divided into remission subgroup and non-remission subgroup. It showed that only estimated glomerular filtration rate (eGFR) between these two subgroups had statistical difference, and eGFR in non-remission subgroup was lower than that in the remission subgroup. Conclusions For high-risk patients,treatment with glucocorticoid combined with immunosuppressant may improve the remission rate of proteinuria significantly.Glomerular filtration rate before treatments is an important prognostic factor.

Effects of Dimethyloxalyl Glycine on ischemic acute renal failure in mice / 中华急诊医学杂志

Objective To explore the effects of Dimethyloxalyl Glycine(DMOG)on isehemic acute renal failure(iARF)in mice and its relationship with activation of hypoxia inducible factor 1α(HIF-1α).Method Twenty five C57/BL male mice were divided into 5 groups randomly:control group,DMOG group,sham operation group,ischemia/reperfusion(I/R)group and DMOG pretreated group(DMOG+I/R).Ischemia/reperfusion injury was induced in mice by clamping both renal pedicles for 30 minutes.The expression of HIF-1α was determined by Western blot.Renal function was reflected by blood urea nitrogen(BUN)and serum creatinine(Scr).Morphologic changes were evaluated under light microscopy.Apoptosis in the kidney was detected by TUNEL staining.Expression of Vimentin,a marker of tubulointerstitial damage was detected by immunohistochemistry.Results The elvated levels of of BUN((65.8±2.6) vs (13.6±0.7),P＜0.01],and Scr[(229.5±11.2) vs (6.5±0.8),P＜0.01]andwere found morphological injury were induced by the ischemic insult in I/R group.Administration of DMOG dramatically improved renal function[BUN,(26.3±6.5)vs(65.8±2.6);Scr,(27.0±14.1)vs(229.5±11.2),P＜0.01]associated with amelioration of tubulointerstital damage.In the DMOG treated group,the protein level of HIF-1α in the kidney of mile was also up-regulated significantly.Conclusions The protection against iARF in mice by DMOG administration is mediated by activation of HIF-1α.

Questionnaire Analysis for International Students Majoring in Medicine Taught in English / 中华医学教育探索杂志

Upon the situation that the scale of international students studying clinic medicine is enlarging,we adopted the questionnaire designed by International Affairs'Office of Ministry of Education to investigate the general information as the reason why international students come to study in China,the entrance standard,the evaluation on teaching,the occupation selecting and the tuition fee etc.The essay aims to provide information for normalizing the education and constituting concerned policy.

Research on the Development Trend of International Students who Pursue their Degree in Medicine / 中华医学教育探索杂志

In allusion to the constant development of degree students majoring in medicine recent years,the essay discusses the future development trend of medicine degree student on analysis of education demand of the neighboring countries and the contrast of different cultivate model and the quality standardization.

The clinical efficacy of capecitabine administrated continuously for three weeks in treating old patients with advanced digestive tract cancer / 中国癌症杂志

0.05) between the control group and the conventional group.Incidences of hand-foot syndrome, nausea, vomiting, stomatitis and diarrhea were significantly lower in the control group in comparison to the conventional group(P