Objective To detect the expression of nesprin-1 gene mRNA and protein and to localize the sub-cellular protein during the development of mouse heart.Methods Samples of embryonic heart tissue were collected from postnatal and adult mice on embryo 12.5 days(E12.5),E14.5,E16.5,and E18.5,respectively.Expression levels of nesprin-1 mRNA and protein and the protein sub-cellular location in samples of embryonic heart tissue from mice were detected by reverse transcriptase PCR,Western blot analysis and immunofluorescence at different time points.Results The expression level of nesprin-1 mRNA and protein in samples of embryonic heart tissue from postnatal and adult mice was measured on E12.5,E16.5 and E18.5,respectively,which increased with the development of mouse heart,reached it peak on E16.5 and E18.5.The lowest expression level of nesprin-1 mRNA and protein was found in adult mice.Immunofluorescence showed that nesprin-1 was distributed in nuclear envelope and in spaces around the nuclei.Conclusion nesprin-1 is dynamically expressed during the development of mouse heart,and highly expressed in the mature period of cardiac conduction system and cardiac muscle cells,indicating that it plays an important role in the development of cardiac conduction system and cardiac muscle cells.

OBJECTIVE To explore a management method for the prevention and control of infectious diseases in hospital. METHODS We applied the PDCA (plan,do,check and action) model to establish a management strategy for the prevention and control of infectious diseases for three years. The components of the management model included providing educational program for medical staff,designing and realizing direct network system for infectious disease reporting,monitoring diagnosed infectious diseases ,analyzing their epidemic tendency,guiding medical staff to prevent and control infectious diseases,and strengthening the surveillance,feedback,rewards and punishment. RESULTS The rate of delayed or missed report of infectious diseases was decreased from 5% to 1.1% by monthly check and from 6.8% to 0% by seasonally cross check. Although 46 cases of AIDS,5 cases of caesarean birth with HIV possitive and 1445 cases of hand-foot-mouth disease were admitted in our hospital,No medical staff and other patients was crossly infected. CONCLUSIONS Our way of management based on PDCA model can strengthen the prevention and control of infectious diseases and assure medical quality and patient safety.

Objective To explore the predictive value of obesity-related index for metabolic syndrome(MS)among Tianjin adults. Methods A total of 522 adults who participated in a physical examination were included in this cross-sectional study. Participants 'gender and age were record-ed,height,weight,waist circumference(WC)and hip circumference were measured,and TC,TG,HDL-C and FPG were tested. The receiver op-erating characteristic(ROC)curves were generated to identify the optimal measurement of obesity for the prediction of metabolic risk in this popu-lation. Results The area under the ROC curve(AUC)value for waist-to-height ratio(WHtR)predicted the maximum value of MS,followed by WC and BMI,and WHR was the lowest. The optimal cut off value of WHtR was 0.509. There was significant difference in the ability of BMI be-tween different age groups and WC to predict MS. In the groups of<40 years old and≥60 years old,BMI had higher predictive value for MS than WC. Conclusion WHtR has the best predictive value for evaluating the MS risk compared to BMI ,WC or WHR alone among Tianjin adults. The predictive value of BMI and WC for evaluating the MS risk can be improved by the method of age stratification.

Objective To analyse the risk factors of vulnerable plaque biomarker and to construct an early warning system. Methods Ninety patients with suspected acute coronary syndrome (ACS) hospitalized during December 2012 and December 2013 were selected. The coronary artery lesions were divided into type I, II and III plaque groups by the morphology of atherosclerotic plaque. Serum SAA, PLGF, sCD40L and Npt were measured. The results of SAA, PLGF, sCD40L and Npt were compared. Logistic regression model was fitted to explore the main influencing factors of the vulnerable plaque. Results SAA, PLGF, sCD40L, and Npt were main influencing factors of the vulnerable plaques, and the ORs were 1.61, 1.88, 1.96 and 1.79 respectively. Conclusion The detection of SAA, PLGF, sCD40L and Npt biochemical markers in patients with chest pain is important for predicting the vulnerable plaque and guiding clinical treatment.

Objective To construct the p300 specific siRNA vector in mice and observe its effect on cardiac transcription factor,GATA4,for further study of the effect of p300-mediated histone acetylation on heart growth. Methods Three recombinant plasmid vectors ( p300 RNAi1,p300 RNAi2,and p300 RNAi3s) ,designed and constructed according to the conservative sequence of mice p300,were respectively tansfected into in vitro cultured cardiac muscle cells of suckling mice. Expression of p300 and GATA4 at mRNA and protein levels was detected by RT-PCR and immunofluorescence,respectively. Results The expression level of p300 mRNA was not significantly decreased in pSOS-HUS and p300 RNAi2 groups,but significantly decreased in p300 RNAi1 and p300 RNAi3 groups ( 0. 220 8 ? 0. 020 0 and 0. 170 8 ? 0. 040 0 vs 0. 509 5 ? 0. 030 0,P

Methods: A total of 10 specific pathogen free (SPF) grade female DBA/2J mice were randomly divided into model group and QGA II group (n = 5 for each group), while additional 5 SPF-grade female C57BL/6J mice were assigned to control group. Mice presented spontaneous high IOP and showed elevated approximately at the age of seven months. The high IOP was maintained until week 38, when gavage was initiated. Mice in control group underwent the same intragastric treatment, while those in QGA II group were gavaged with QGA II (9.67 g/kg), once a day for four weeks. Retinal morphology was examined using hematoxylin and eosin (HE) staining, with the number of retinal ganglion cells (RGCs) counted. The expression level of Brn3a protein, a specific marker for RGCs, was detected by immunofluorescence, with the mean optical density (OD) measured for quantitative analysis. In addition, 16S rDNA sequencing was leveraged to analyze changes in the diversity of gut microbiota, including their α-diversity (Chao1, Shannon, Pielou’s evenness, and observed species index) and β-diversity. Venn diagrams and linear discriminant analysis effect size (LEfSe) analysis was employed to investigate the number of amplicon sequence variants (ASVs), the abundance of differential gut microbiota species, and the classification of species at both the phylum and genus levels within the three groups of mice. Results: HE staining revealed that compared with control group, model group showed significant reduction in the number of RGCs (P < 0.01), with intracellular vacuolar degeneration and nuclear pyknosis. After QGA II treatment, the number of RGCs was significantly increased compared with model group (P < 0.01), with notable improvements in intracellular vacuolar degeneration. Immunofluorescence analysis showed that the mean OD of Brn3a protein was significantly decreased in model group compared with control group (P < 0.01), while QGA II treatment significantly elevated its expression level (P < 0.01). Analysis of α-diversity showed that after QGA II intervention, the Chao1, Shannon, and Pielou’s evenness indices were significantly increased (P < 0.01), and the observed species index was elevated (P < 0.05). β-Diversity analysis demonstrated distinct clustering among the three groups, indicating relatively low similarity in bacterial community structures. ASV clustering identified a total of 14 061 ASVs across all groups, with 9 514 ASVs shared between model and QGA II groups. At the phylum level, the abundance of Bacteroidetes was significantly decreased in model group compared with control group (P < 0.01), while Firmicutes and the Firmicutes/Bacteroidetes (F/B) ratio were significantly increased (P < 0.01). QGA II treatment significantly reduced both Firmicutes abundance and the F/B ratio (P < 0.01). At the genus level, Lactobacillus was dominant across all groups, with its abundance significantly increased in model group (P < 0.01) and subsequently decreased following QGA II intervention (P < 0.05). Conclusion QGA II restructured the gut microbiota of DBA/2J mice with chronic high IOP, bringing changes in their diversity and abundance of components. Firmicutes, Bacteroidetes, Lactobacillus, along with their associated microorganisms, are likely critical components of the gut microbiota that contribute to the optic neuroprotective effects of QGA II on chronic high IOP mice.

Background Researches showed that the increase of intraocular pressure (IOP) in glaucomatous eye is associated with the increasing resistance to aqueous humor outflow effects of transforming growth factor-β (TGF-β) and CD44.Qingguangan is a traditional Chinese medicine and used to treat glaucoma.However,its mechanism of lowing-IOP effect is not elucidated.Objective This study was to investigate the lowing-IOP effect and mechanism of qingguangan granule in DBA/2J mouse,a spontaneous glaucoma model mice.Methods Ten 3 month-old female DBA/2J mice with normal IOP were chosen as control group,and 20 spontaneous ocular hypertension mice aged 9 months were randomized into high IOP group and qingguangan-treated group,with 10 mice for each group.The qingguangan (2.5 g/kg) was administered by gavaging twice per day for consecutive 15 days in the qingguangan-treated group,and normal saline solution was used in the same way in the control group and high IOP group.IOP was measured by anterior chamber injection/suction system at a perfusion rate of 2.5 and 5.0 μl/min,respectively,and the coefficient of aqueous outflow facility (C value) and outflow resistance (R value) were calculated.Another 60 3-month-old DBA/2J mice were randomized into blank control group gavaged with normal saline solution and high-,middle-and low-dose qingguangan groups gavaged with 25.00,12.50 and 6.25 g/kg drugs,respectively,and the mouse serum containing drugs was extracted 7 days after treatment.The scleral tissue with trabecular meshwork were obtained for the culture of trabecular meshwork cells and the cells were identified by immunohistochemistry of fibronectin (FN),laminin (LN) and neuronspecific enolase (NSE).TGF-β was added into the medium for 24 hours with the final concentration of 0,5,10,20,50 and 100 ng/ml,and MMT chromatometry was employed to detect the cell vitality.The cells pre-treated with 20 ng/ml TGF-β were treated with different concentration of drug serum for 24,48 and 72 hours,and the level of TGF-β2 receptor in cell supernatant and the expression of CD44 protein in the cells were detected by ELISA and Western blot assay,respectively.Results The IOPs with perfusion both 2.5 μl/min and 5.0μl/min in the qingguangan-treated group and the control group were significantly lower than those in the high IOP group (all at P＜0.01).Compared with the high IOP group,the C value was significantly reduced (2.35±1.34 vs.1.08±0.36) and the R value was evidently elevated (0.64±0.55 vs.1.05± 0.47) in the qingguangan-treated group (all at P＜0.01).Cultured cells were spindle-shaped with the positive response to FN,LN and NSE antibody.The cell vitality was lower in the 5,10 and 20 ng/ml TGF-β group than that in the 0 ng/ml TGF-β group (all at P＜0.05).Compared with the blank control group,the TGF-β2 receptor content in the supernatant and the related expression level of CD44 protein in the cells were elevated in the TGF-β-treated group (all at P＜0.01),and TGF-β2 receptor contents and CD44 expression levels in the TGF-β+high dose drug serum group was significantly lower than those in the TGF-β group and TGF-β +low dose drug serum group 24,48 and 72 hours after culture (all at P＜0.01).Conclusions Qingguangan can lower IOP of spontaneous glaucoma mice by affecting aqueous humor dynamics.Serum containing qingguangan down-regulates the expressions of TGF-β2 receptors and CD44 in trabecular meshwork cells in vitro.

This paper summarized PENG Qinghua's experience in treating age-related dry eye based on the theory of ministerial fire. Guided by the theory of ministerial fire， it is believed that the key pathogenesis of age-related dry eye is transgression of ministerial fire， and kidney yang deficiency-cold， kidney yin depletion， and spleen-earth insufficiency are the causes of transgression of ministerial fire. The general principle of treatment is to conceal the ministerial fire. For kidney yang deficiency-cold and floating ministerial fire， the treatment should tonify the kidney and warm yang， and return fire to its origin， with Yin Huo Decoction （引火汤）； for kidney yin depletion and frenetic stirring of ministerial fire， the treatment should tonify the kidney and supplement essence， enrich yin and reduce fire， with Sancai Fengsui Pellet （三才封髓丹）； and for spleen-earth insufficiency， and fumigation of ministerial fire， the treatment should strengthen and activate spleen qi， and reinforce soil and hidden fire， with Renshen Huangqi Decoction （人参黄芪汤）.

Objective:To recognize the development pattern of COVID-19 epidemic, and to predict the development trend of the epidemic in three categories: the country, Hubei province, and areas outside Hubei province.Methods:The BloComp model was used to fit existing data. With the estimated parameters, we simulated future epidemic development.Results:The pattern of virus transfer among different populations in Hubei province was significantly different from that outside Hubei province. It is predicted by the BloComp model that the time when the number of confirmed cases fell to zero in Hubei province and areas outside Hubei will be May 2020 and April 2020, respectively.Conclusions:The epidemic situation will end at the end of May 2020. Adequate medical resources investment and more comprehensive and accurate investigation of suspected people play important roles in controlling the epidemic situation.

Objective To explore the age-related biological properties of bone-derived mesenchymal stem cells(BMSCs)from mice of different age groups and their osteogenic differentiation induced by bone morphogenetic protein 2(BMP2).Methods Eight C57BL/6J mice were divided into a young group(4 weeks old,weighing 10～15 g,n=4)and an old group(12 months old,weighing 20～25 g,n=4),with half male and half female.MSCs were extracted from the whole bones of the 2 groups of mice.After the obtained cells were identified with flow cytometry for the surface markers,β-galactosidase staining was employed to compare the senescence level of BMSCs,MTT and EdU incorporation assays were conducted to compare the proliferation and self-renewal abilities of between the 2 groups.Western blotting was employed to analyze the expression of CyclinD1 and P21 in BMSCs.Then ALP staining,Alizarin Red staining and RT-qPCR were used to evaluate the osteogenic differentiation ability of the cells.RNA sequencing was performed to compare the differential gene expression in BMP2-induced BMSCs.Lastly,the sequencing results were re-confirmed by using flow cytometry.Results Flow cytometry showed that the sorted and cultured mouse BMSCs met the criteria for MSCs.The results of β-galactosidase staining indicated that the senescence level of BMSCs in the old group was significantly higher than that in the young group(P＜0.05).MTT and EdU doping experiments revealed that the cell viability and proliferation ability of BMSCs were significantly lower in the old group than the young group(P＜0.05).Western blotting displayed that the expression level of cell cycle protein CyclinD1 was lower,whereas that of cell cycle inhibitory factor P21 was significantly higher in the BMSCs from the old group than the cells from the young group(P＜0.05).ALP/Alizarin Red staining and RT-qPCR demonstrated that the BMSCs from the young group had stronger osteogenic differentiation capacity after BMP2 treatment when compared the cells of the old group(P＜0.05).RNA sequencing results displayed that the changing profile of CD51 expression was in opposite trends in the young and old BMSCs after BMP2 treatment.Finally,flow cytometry revealed that the percentage of CD51+cells within the CD45-cells was significantly higher in the young group than the old group.Conclusion The decrease in the percentage of CD51+cells among CD45-cells in aged BMSCs is closely associated with their decreased responsiveness to BMP2-induced osteogenic differentiation.