AIM:The objective of this study is to examine the expression of profilin 1(PFN1)in mice with di-abetic nephropathy and determine its association with immune cell infiltration.METHODS:This study presents an analy-sis of PFN1 expression and immune cell infiltration in patients with diabetic nephropathy,utilizing transcriptome expres-sion data from kidney tissue microarray.Additionally,the findings were validated in a diabetic nephropathy mouse model.Sixteen C57BL/6 mice were randomly assigned into two groups,namely the normal group and the model group,in an equal manner.The model group underwent the establishment of the diabetic nephropathy model through intraperitoneal injection of streptozotocin.Subsequently,the expression levels of CD11b,F4/80,CC chemokine receptor 4(CCR4),interleukin-1 receptor type I(IL-1R1),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-3 in kidney tissue were assessed upon successful establishment of the diabetic nephropathy model.Furthermore,the overexpression of PFN1 was observed in a cellular model of diabetic nephropathy,and the protein expression levels of monocyte chemotactic pro-tein-1(MCP-1)and caspase-3 were assessed.RESULTS:The expression of PFN1 was found to be significantly in-creased in the GSE30122 dataset of transcriptome expression in kidney tissues affected by diabetic nephropathy(P＜0.01).This increase in PFN1 expression was found to be correlated with the presence of macrophages and T cells.Fur-thermore,the renal tissue of the diabetic nephropathy model group exhibited significant pathological changes.In this mod-el group,the expression levels of PFN1,CD11b,F4/80,CCR4,IL-1R1,Bax,Bcl-2,and caspase-3 were all significant-ly increased(P＜0.01).Overexpression of PFN1 could enhance the expression of MCP-1 and caspase-3 proteins.CON-CLUSION:Macrophages and Th17 cells were identified within the renal tissue of mice with diabetic nephropathy,con-comitant with an up-regulation in the expression of PFN1.This up-regulation was observed to facilitate the induction of apoptosis in the context of diabetic nephropathy.

OBJECTIVE： To explore the component， target and pathway of Panax notoginseng for coronary heart disease （CHD） and its potential molecular mechanism. METHODS： Based on network pharmacology， active components of P. notoginseng were retrieved with TCMSP platform. The targets of P. notoginseng for CHD were screened by using DRAR-CPI server， GeneCards and DisGeNET databases. Cytoscape 3.6.0 software was used to form the effective components-CHD targets network of P. notoginseng. String database was used to draw target interaction network. Network Analyzer tool was used to calculate target connectivity， and potential core targets were screened. Molecular docking between the core targets and the effective components of P. notoginseng was performed by Systems Dock Web Site server. KEGG pathway enrichment analysis and gene ontology （GO） enrichment analysis were also carried out to explore the important signal pathway and molecular function of P. notoginseng for CHD. “Effective component-target-signal pathway”network of important signal pathway were constructed. RESULTS： Five effective components （stigmasterol， β-sitosterol， ginsenoside rh2， quercetin， notoginsenoside r1） were screened from P. notoginseng for CHD， which acted on 96 targets and had 134 functional relationships. Five core targets were protein kinase B （AKT）， interleukin 6 （IL-6）， vascular endothelial growth factor A （VEGFA）， c-JUN protein （c-JUN） and heparin binding epidermal growth factor （HB-EGF）， which played an important role in the treatment of CHD by altering protein binding and regulating signaling pathways as phosphatidylinositol-3 kinase-protein/kinase B （PI3K/AKT）， hypoxia-inducible factor-1 （HIF-1） and mitogen-activated protein kinase （MAPK）. CONCLUSIONS： P. notoginseng in the treatment of CHD is not only play a variety of effects through the role of multiple targets， but also produce complex network regulation effect through the interaction between targets.

Objective:To investigate the effect of enteral nutrition support by CT-guided percutaneous gastrostomyon the nutritional status of patients with esophageal cancer complicated with dysphagia during radiotherapy.Methods:Therewere46 cases of esophageal cancer patients with dysphagia treated with CT-guided percutaneousgastrostomy.Others 43 cases of esophageal cancer by oral feeding in patients with dysphagia as control groupduringthe sametimein our hospital radiotherapy center.Patients in the observation group were ingested daily through the gastrostomy,and the nutritional intake of the control group included oral ingestion and intravenous infusion.All patients were measuredthe body height,body weight (BW).body mass index (BMI),Serum levels of serum albumin (ALB),pre-albumin (PA) and hemoglobin (HB) before and after radiotherapy.We also observed the incidence of acute radiation esophagitis and the completion of the treatment plan during radiotherapy in both groups,and to observe the two groups of patients the incidence rate of radiotherapy and treatment plan during the completion of acute radiation esophagitis.Results:There was no significant difference in BW,BMI,ALB,PA,HB before radiotherapy between the two groups (t =0.84,0.63,-1.07,-0.81,1.48,P ＞ 0.05).The BW,BMI,ALB,PA and HB of the observation group were significantly higher than those of the control group at the end of radiotherapy,which werestatistically significant (t=3.30,4.65,6.82,43.56,31.91,P ＜ 0.01).During the radiotherapy,the total incidence of acute radiation esophagitis in the observation group was significantly lower than that in the control group,(x2=3.971,P＜ 0.05).In addition,the completion rate of the observationgroup was significantly higher than that of the control group (x2 =6.811,P ＜ 0.01).Conclusion:To the Patients with dysphagia of esophageal cancer,enteral nutrition byCT guided percutaneous gastrostomy,can improve the malnutrition,the immune function of the patients and reduce the acute radiation esophagitis during radiotherapy and ensure the successful completion of the treatment plan.

Objective To explore the effect of follow-nursing model in the nutritional conditions of allogeneic hematopoietic stem cell transplantation(allo-HSCT)patients. Methods Eighty patients who underwent allo- HSCT from January 2015 to December 2016 were enrolled and divided into the observation group(40 cases)and the control group(40 cases)by random digit table.The observation group was treated with follow-nursing model, while the control group was treated with regular nursing. The Subjective Global Assessment (SGA), hematologic indexes and other body measurement indexes of the two cohorts were assessed three months later. Results The incidence of malnutrition in the observation group was 52.5%(21/40),compared to the control group with 77.5%(31/40)(χ2=4.451,P<0.05).The indexes including body mass index, triceps skin fold, arm muscle circumference, albumin, prealbumin and hemoglobin in the observation group were obviously higher than the control group with statistical significance (t=2.599-36.481, P<0.05 or 0.01). Conclusions The follow-nursing model may improve the nutritional conditions and some other body indexes of allo-HSCT patient compared to regular nursing model.

OBJECTIVETo examine the expression profile and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum (Sjp40) during granuloma formation in the liver of infected New Zealand white rabbits.

METHODSNew Zealand white rabbits were infected with S. japonicum cercariae, and the livers were harvested at 29 and 45 days post-infection (dpi). The total RNA of the liver tissues was extracted for expression profiling of Sjp40 by quantitative reverse transcription-PCR (qRT-PCR) with GAPDH of S. japonicum as the endogenous reference gene. The expression of Sjp40 in the liver were detected by Western blotting using anti-Sjp40 monoclonal antibody (mAb) 9G7 or anti-Toxoplasma gondii tSAG1 mAb Y3A8 (control) as the primary antibody. Paraffin sections of the liver were prepared for observing egg granuloma formation using HE staining and for indirect immunofluorescence assay of Sjp40 location in the trapped eggs and egg granulomas.

RESULTSThe level of Sjp40 mRNA in the eggs trapped in rabbit livers was significantly higher at 45 dpi than that at 29 dpi (P<0.05), and Western blotting confirmed the presence of Sjp40 protein in the rabbit livers at both 29 and 45 dpi. Immunofluorescence assay demonstrated localized expression of Sjp40 in the immature eggs in the rabbit liver at 29 dpi, but at 45 dpi fluorescence was detected in clusters of mature eggs containing miracidium and in the surrounding egg granulomas.

CONCLUSIONSThe transcriptional levels of Sjp40 significantly increased with the maturation of eggs trapped in the rabbit livers. Sjp40 protein spread from the eggs to the surrounding egg granuloma at 45 dpi when acute liver granulomatous lesions occur, suggesting that Sjp40 plays a key role in egg granulomas formation in the livers of infected New Zealand white rabbits.

Animals ; Antibodies, Monoclonal ; Antigens, Helminth ; metabolism ; Fluorescent Antibody Technique ; Gene Expression Profiling ; Granuloma ; parasitology ; Helminth Proteins ; metabolism ; Liver ; parasitology ; RNA, Messenger ; Rabbits ; Schistosoma japonicum ; Schistosomiasis japonica

Objective To examine the expression profile and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum (Sjp40) during granuloma formation in the liver of infected New Zealand white rabbits. Methods New Zealand white rabbits were infected with S. japonicum cercariae, and the livers were harvested at 29 and 45 days post-infection (dpi). The total RNA of the liver tissues was extracted for expression profiling of Sjp40 by quantitative reverse transcription-PCR (qRT-PCR) with GAPDH of S. japonicum as the endogenous reference gene. The expression of Sjp40 in the liver were detected by Western blotting using anti-Sjp40 monoclonal antibody (mAb) 9G7 or anti-Toxoplasma gondii tSAG1 mAb Y3A8 (control) as the primary antibody. Paraffin sections of the liver were prepared for observing egg granuloma formation using HE staining and for indirect immunofluorescence assay of Sjp40 location in the trapped eggs and egg granulomas. Results The level of Sjp40 mRNA in the eggs trapped in rabbit livers was significantly higher at 45 dpi than that at 29 dpi (P<0.05), and Western blotting confirmed the presence of Sjp40 protein in the rabbit livers at both 29 and 45 dpi. Immunofluorescence assay demonstrated localized expression of Sjp40 in the immature eggs in the rabbit liver at 29 dpi, but at 45 dpi fluorescence was detected in clusters of mature eggs containing miracidium and in the surrounding egg granulomas. Conclusion The transcriptional levels of Sjp40 significantly increased with the maturation of eggs trapped in the rabbit livers. Sjp40 protein spread from the eggs to the surrounding egg granuloma at 45 dpi when acute liver granulomatous lesions occur, suggesting that Sjp40 plays a key role in egg granulomas formation in the livers of infected New Zealand white rabbits.

Objective To examine the expression profile and immunofluorescence localization of the major egg antigen p40 of Schistosoma japonicum (Sjp40) during granuloma formation in the liver of infected New Zealand white rabbits. Methods New Zealand white rabbits were infected with S. japonicum cercariae, and the livers were harvested at 29 and 45 days post-infection (dpi). The total RNA of the liver tissues was extracted for expression profiling of Sjp40 by quantitative reverse transcription-PCR (qRT-PCR) with GAPDH of S. japonicum as the endogenous reference gene. The expression of Sjp40 in the liver were detected by Western blotting using anti-Sjp40 monoclonal antibody (mAb) 9G7 or anti-Toxoplasma gondii tSAG1 mAb Y3A8 (control) as the primary antibody. Paraffin sections of the liver were prepared for observing egg granuloma formation using HE staining and for indirect immunofluorescence assay of Sjp40 location in the trapped eggs and egg granulomas. Results The level of Sjp40 mRNA in the eggs trapped in rabbit livers was significantly higher at 45 dpi than that at 29 dpi (P<0.05), and Western blotting confirmed the presence of Sjp40 protein in the rabbit livers at both 29 and 45 dpi. Immunofluorescence assay demonstrated localized expression of Sjp40 in the immature eggs in the rabbit liver at 29 dpi, but at 45 dpi fluorescence was detected in clusters of mature eggs containing miracidium and in the surrounding egg granulomas. Conclusion The transcriptional levels of Sjp40 significantly increased with the maturation of eggs trapped in the rabbit livers. Sjp40 protein spread from the eggs to the surrounding egg granuloma at 45 dpi when acute liver granulomatous lesions occur, suggesting that Sjp40 plays a key role in egg granulomas formation in the livers of infected New Zealand white rabbits.

Objectives To investigate gestational multiple metabolic abnormalities aggregation and diagnostic criteria for gestational metabolic syndrome(GMS),and to analyze the risk factors of GMS.Methods A cohort study recruiting 309 pregnant women with preeclampsia,627 pregnant women with gestational diabetes mellitus(GDM)and 1245 normal pregnant women was performed from January 2008 to December 2011 in Guangdong Women and Children's Hospital.Information regarding age,gestational weeks,basic blood pressure,admission blood pressure,height and body mass index(BMI)before pregnancy was recorded.Biochemical indicators including fasting plasma glucose(FPG),fasting insulin (FINS),total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),free fatty acids(FFA)were tested.GMS was diagnosed with three or all of the following conditions:(1)overweight and/or obesity before pregnancy(BMI ≥ 25 kg/m2);(2)hypertension with blood pressure ≥ 140/90 mm Hg(1 mm Hg =0.133 kPa);(3)hyperglycemia:diagnosed as GDM;(4)dyslipidemia with TG≥3.23 mmol/L The incidence of GMS of the three groups were calculated and the risk factors were analyzed.Results(1)The age,gestational weeks,basic blood pressure,admission blood pressure,BMI before pregnancy of women with preeclampsia and women with GDM were significantly different compared to normal women,respectively(P ＜ 0.01).(2)Biochemical indicators of women with preeclampsia were as following:FPG(4.6 ± 1.0)mmol/L,FINS(10.1 ± 5.6)mU/L,TC(6.3 ±1.6)mmol/L,TG(3.9 ± 1.8)mmol/L,HDL-C(1.4 ±0.4)mmol/L,LDL-C(3.0 ± 1.0)mmol/L,FFA (0.8 ±0.4)mmol/L.And those in women with GDM were:FPG(4.7 ± 0.9)mmoL/L,FINS(10.2 ± 5.8)mU/L,TC(5.7 ± 1.3)mmol/L,TG(3.2 ± 1.1)mmol/L,HDL-C(1.4 ± 0.4)mmol/L,LDL-C (2.7 ± 0.9)mmol/L,FFA(0.6 ± 0.3)mmol/L In normal pregnant women they were:FPG(4.3 ±0.5)mmol/L,FINS(9.0±4.4)mU/L,TC(5.7 ±1.1)mmol/L,TG(2.8 ±1.1)mmol/L,HDL-C (1.5 ± 0.4)mmol/L,LDL-C(2.9 ± 0.8)mmol/L,FFA(0.6 ± 0.2)mmol/L Statistic differences were found in preeclampsia and GDM women compared to normal women respectively(P ＜ 0.01).(3)The prevalence of GMS in preeclampsia group and in GDM group was 26.2％(81/309)and 13.6％(85/627),statistically different from that of the control group(0)(P ＜0.01).(4)Compared to normal women,women with preeclampsia had higher risk of developing GMS(OR =1.62,95 ％ CI 1.31-2.00,P ＜ 0.01).The risk factors were BMI(OR =1.29,95％ CI 1.13-1.47)and TG(OR =2.49,95％ CI 1.87-3.31).Also,women with GDM had higher risk of developing GMS than normal women(OR =1.27,95％ CI 1.09-1.49,P ＜ 0.01),and the risk factors were BMI(OR =1.13,95 ％ CI 1.04-1.23)and TG(OR =1.16,95 ％ CI 1.02-1.33).TG was the independent risk factor in both preeclampsia women and GDM women(P ＜ 0.01,P ＜ 0.05).HDL-C seemed to have less importance in identifying GMS(P ＞ 0.05).Conclusions According to the GMS diagnostic criteria used in this study,some preeclampsia patients and some GDM women had aggregation of multiple metabolic abnormalities including pre-pregnancy overweight/obesity,hyperglycemia,high blood pressure and dyslipidemia.TG was the independent risk factor for GMS.HDL-C seemed to have less importance in identifying GMS.

OBJECTIVETo develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions.

METHODSTo establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA.

RESULTSOur data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration.

CONCLUSIONWe have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.

Gene Expression Regulation, Plant ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Hydroponics ; methods ; Plants, Genetically Modified ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Tobacco ; genetics ; growth & development

OBJECTIVETo study the effect of RNA interference (RNAi) on small heat shock protein (sHSP) Sjp40 of Schistosoma japonicum and its synergistic effect on the expression of SjHSP60, SjHSP70, and SjHSP90 mRNA, and observe the mRNA expression levels of Sjp40, SjHSP60, SjHSP70, and SjHSP90 in different stages of S.japonicum.

METHODSDouble-stranded RNA (dsRNA) of Sjp40 (dsSjp40) and a control dsRNA of green fluorescent protein (dsGFP) were generated by in vitro transcription and transfected into adult worm by immersing the worm in dsRNA solution. The total RNA and proteins were isolated simultaneously from the adult worms using TRIzol reagent 7 days after transfection. The expression levels of Sjp40, SjHSP60, SjHSP70, and SjHSP90 mRNA and the expression level of Sjp40 protein were determined by quantitative real-time PCR (qPCR) and Western blotting, respectively. The mRNA expression of HSPs of S. japonicum in different stages was evaluated by qPCR.

RESULTSCompared with those in the control worms transfected with dsGFP, Sjp40 mRNA level was decreased by 80% in the worms transfected with dsSjp40, and the level of Sjp40 protein showed also a significant decrease. The mRNA expression levels of SjHSP60, SjHSP70, and SjHSP90 did not show an obvious synergism after Sjp40 RNAi. The expression profiles of Sjp40, SjHSP60, SjHSP70, and SjHSP90 showed significant differences in different stages of S. japonicum, and the expression level of Sjp40 mRNA in the egg stage was much higher than that of other HSP genes.

CONCLUSIONdsSjp40-RNAi can induce effective suppression of Sjp40 gene expression at both mRNA and protein levels, but no obvious synergism occurs in the mRNA expressions of SjHSP60, SjHSP70, and SjHSP90.

Animals ; Gene Expression Profiling ; Heat-Shock Proteins, Small ; genetics ; Helminth Proteins ; genetics ; RNA Interference ; RNA, Double-Stranded ; RNA, Messenger ; genetics ; Schistosoma japonicum ; genetics