Objective To explore the regulatory effect of long non-coding RNA HOX transcript anti-sense RNA(lnRNA HOTAIR)targeting tuberous sclerosis complex 1(TSC1)on podocyte injury in diabetic nephropathy(DN).Methods Mice podocyte MPC5 were divided into 6 groups:the control group,the high glucose group(the HG group),the HG+si-NC group,the HG+si-HOTAIR group,the HG+si-HOTAIR+sh-NC group,and the HG+si-HOTAIR+sh-TSC1 group.qPCR was used to detect the levels of lnRNA HO-TAIR and TSC1 mRNA.Cell viability was detected by cell counting kit-8(CCK-8),and apoptosis was detec-ted by flow cytometry.ELISA was used to detect the levels of inflammatory factors[IL-6,tumor necrosis fac-tor-α(TNF-α)]and the levels of oxidative stress indicators[reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD)]were detected by corresponding reagent kit.Results Compared with the control group,the levels of lnRNA HOTAIR,apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG group were significantly increased(P<0.05),while the levels of TSC1 mRNA,cell viability and SOD were significantly decreased(P<0.05).Compared with the HG+si-NC group,the levels of lnRNA HOTAIR,ap-optosis rate,IL-6,TNF-α,ROS and MDA in the HG+si-HOTAIR group significantly decreased(P<0.05),the levels of TSC1 mRNA,cell viability and SOD were significant increased(P<0.05).Compared with the HG+si-HOTAIR+sh-NC group,the levels of apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG+si-HOTAIR+sh-TSC1 group were significantly increased(P<0.05),the levels of TSC1 mRNA,cell viability and SOD were significantly decreased(P<0.05).Conclusion Silencing lnRNA HOTAIR can target TSC1 to regulate the activity,apoptosis rate,inflammatory level and oxidative stress levels of podocyte injury in DN,thereby alleviating podocyte damage in DN.

BACKGROUND:The regulatory mechanisms of acupotomy intervention for knee osteoarthritis at a transcriptome level are not well understood despite its proven clinical efficacy.OBJECTIVE:Using acupotomy therapy in a rat model of knee osteoarthritis,to conduct transcriptome sequencing and bioinformatics analysis on cartilage samples,along with validation,and to reveal the molecular regulatory mechanisms involved in this therapy for knee osteoarthritis in rats.METHODS:Forty-eight Sprague-Dawley rats were selected and divided into three groups by random number table method,acupotomy group,model group,and sham operation group,with 16 rats in each group.Osteoarthritis models were induced by medial meniscus instability in the acupotomy group and model group.After successful modeling,acupotomy group rats were treated with acupotomy once a week,for 4 weeks in total.After the intervention,cartilage samples from the rat's knee joint were stained with hematoxylin-eosin staining and safranin O-fast green staining,evaluated by Mankin scores,and underwent MicroCT scanning.Serum inflammatory factor levels were detected by Elisa.Transcriptome sequencing was performed on the remaining cartilage samples,and the data were analyzed using R software to identify differential gene expression levels among the groups.Core targets were screened through protein-protein interaction network and Cytoscape and validated using RT-qPCR.RESULTS AND CONCLUSION:Compared with the sham operation group,rats in the model group had rough and uneven articular cartilage surfaces,narrowed joint spaces,destroyed articular surface structures,elevated Mankin scores,and significant increases in serum levels of interleukin-6,tumor necrosis factor-α,and matrix metalloproteinase 13(P<0.05).Compared with the model group,rats in the acupotomy group had smoother articular cartilage surfaces,wider joint spaces,slightly irregular articular surfaces,lower Mankin scores,and significantly lower serum levels of interleukin-6,tumor necrosis factor-α,and matrix metalloproteinase-13(P<0.05).Gene ontology and Kyoto genome and genome encyclopedia analyses involved proteolytic metabolism,autophagy,mitogen-activated protein kinase,nuclear factor kB,and Wnt signaling pathways.Protein-protein interaction network and Cytoscape screened for four key genes,including ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor.The mRNA expression of ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor in the articular cartilage of rats in the model group was lower than that of the sham operation group(P<0.05),while the mRNA expression of ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor in the articular cartilage of rats in the acupotomy group was higher than that in the model group(P<0.05).To conclude,acupotomy treatment of knee osteoarthritis in rats may act on signaling pathways such as MAPK,nuclear factor kB,and Wnt to promote cartilage repair,and is closely related to the expression of genes associated with ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor.

BACKGROUND:The regulatory mechanisms of acupotomy intervention for knee osteoarthritis at a transcriptome level are not well understood despite its proven clinical efficacy.OBJECTIVE:Using acupotomy therapy in a rat model of knee osteoarthritis,to conduct transcriptome sequencing and bioinformatics analysis on cartilage samples,along with validation,and to reveal the molecular regulatory mechanisms involved in this therapy for knee osteoarthritis in rats.METHODS:Forty-eight Sprague-Dawley rats were selected and divided into three groups by random number table method,acupotomy group,model group,and sham operation group,with 16 rats in each group.Osteoarthritis models were induced by medial meniscus instability in the acupotomy group and model group.After successful modeling,acupotomy group rats were treated with acupotomy once a week,for 4 weeks in total.After the intervention,cartilage samples from the rat's knee joint were stained with hematoxylin-eosin staining and safranin O-fast green staining,evaluated by Mankin scores,and underwent MicroCT scanning.Serum inflammatory factor levels were detected by Elisa.Transcriptome sequencing was performed on the remaining cartilage samples,and the data were analyzed using R software to identify differential gene expression levels among the groups.Core targets were screened through protein-protein interaction network and Cytoscape and validated using RT-qPCR.RESULTS AND CONCLUSION:Compared with the sham operation group,rats in the model group had rough and uneven articular cartilage surfaces,narrowed joint spaces,destroyed articular surface structures,elevated Mankin scores,and significant increases in serum levels of interleukin-6,tumor necrosis factor-α,and matrix metalloproteinase 13(P<0.05).Compared with the model group,rats in the acupotomy group had smoother articular cartilage surfaces,wider joint spaces,slightly irregular articular surfaces,lower Mankin scores,and significantly lower serum levels of interleukin-6,tumor necrosis factor-α,and matrix metalloproteinase-13(P<0.05).Gene ontology and Kyoto genome and genome encyclopedia analyses involved proteolytic metabolism,autophagy,mitogen-activated protein kinase,nuclear factor kB,and Wnt signaling pathways.Protein-protein interaction network and Cytoscape screened for four key genes,including ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor.The mRNA expression of ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor in the articular cartilage of rats in the model group was lower than that of the sham operation group(P<0.05),while the mRNA expression of ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor in the articular cartilage of rats in the acupotomy group was higher than that in the model group(P<0.05).To conclude,acupotomy treatment of knee osteoarthritis in rats may act on signaling pathways such as MAPK,nuclear factor kB,and Wnt to promote cartilage repair,and is closely related to the expression of genes associated with ataxia-telangiectasia mutations,Myb SWIRM and MPN domain protein 1,heat shock protein 90α1,and NIPBL cohesion-loading factor.

Amyloidosis is a metabolic disease characterized by the deposition of amyloidogenic fibrils in multiple organs, which causes structural disorders, leading to a decline in organ function and in the case of myocardial involvement, damage to the structure and function of the heart, which is the main cause of mortality. However, the disease is often misdiagnosed or underdiagnosed due to its insidious onset and lack of specificity in clinical manifestations. 99Tc m-pyrophosphate (PYP) SPECT is a noninvasive test that can not only identify cardiac amyloid deposits at an early stage but also provide a useful tool for the diagnosis of cardiac amyloidosis. This article briefly describes the traditional screening methods for cardiac amyloidosis, outlines their limitations, and focuses on the clinical diagnostic value of 99Tc m-PYP SPECT in cardiac amyloidosis.

Objective:Objective The inhibitory effect of Niuhuang Qinggan Capsule on 2019-nCoV virus was studied to provide experimental evidence for drug screening and treatment of 2019-nCoV acute respiratory disease.Methods:The 2019-nCoV virus GS048 strain was isolated and cultured by Vero-E6 cells. The solution of Niuhuang Qinggan capsule was incubated with virus in vitro to determine the change of virus titer. The effect of Niuhuang Qinggan capsule on the proliferation of 2019-nCoV virus was determined by cell culture medium method.Results:It was found that the concentration of 274.3 μg/ml was not toxic to the cells when the concentration of drugs was lower. When the drug was incubated with virus, the titer of virus decreased from 10 4.4TCID 50/ml to 10 2.8TCID 50/ml. In vitro 2019-nCoV virus culture experiment showed that the addition of the drugs reduced the cytopathy, and the logarithmic value of virus titer decreased by 40% -50%. Conclusion:In this study, in vitro inhibition experiments suggest that Niuhuang Qinggan capsule has the potential to inhibit the proliferation of 2019-nCoV. It can be used as a candidate drug to further verify the therapeutic effect of the 2019-nCoV acute respiratory disease.

Objective:To investigate the cultural construction of Traditional Chinese Medicine(TCM) hospitals in Hunan province, for reference in their cultural construction.Methods:A questionnaire was designed according to TCM Hospital Cultural Construction Guidelines by the National Traditional Chinese Administration.From May to June 2020, the questionnaire was used to survey the present situation, existing problems and development suggestions of TCM cultural construction at TCM hospitals of and above the county level in Hunan province. The data were analyzed by descriptive analysis.Results:The survey covered 117 public TCM hospitals in the province and 87 questionnaires were recovered, a rate of 74.4%. In terms of cultural construction in these institutions, various programs were in place at a relatively high proportion in the core cultural value construction. The development of behavioral norms teaching and inheritance was in place at a relatively low proportion, namely 78.8% at tertiary TCM hospitals, 51.8% at secondary TCM hospitals, and only 16.7% at secondary level-B TCM hospitals. In terms of environmental image construction, various programs were in place at a relatively high proportion, yet focusing on themed cultural wall posters culturol bulletin board and other forms of display. At present, the main problems were insufficient funding(86.9%) and talents(82.1%).Conclusions:TCM hospitals in Hunan province highly value the construction of cultural core value and environmental image, but their development in the code of conduct system and other connotation was weak.

AIM: To investigate the apoptosis and molecular mechanism of human hepatocellular carcinoma HepG2 cells induced by ginsenoside Rh4.METHODS: Human hepatocellular carcinoma HepG2 cells were treated with ginsenoside Rh4 at doses of 10, 20 and 40 μmol/L, and the inhibitory effect of ginsenoside Rh4 on HepG2 cell viability was measured by MTT assay.The apoptotic rate of HepG2 cells was analyzed by flow cytometry.The morphological changes of the HepG2 cells were observed by Hoechst 33258 and TUNEL staining.The expression of apoptosis-related proteins Bax, Bcl-2, caspase-3 and caspase-9 was determined by Western blot.RESULTS: Ginsenoside Rh4 promoted apoptosis of HepG2 cells in a dose-dependent manner.TUNEL and Hoechst 33258 staining showed that the cells appeared obvious shrinking, swelling and rupture after treated with ginsenoside Rh4 for 24 h.The results of Western blot showed that with the increasing concentrations of ginsenoside Rh4, the expression of pro-apoptotic proteins Bax, cleaved caspase-3 and caspase-9 increased, while anti-apoptotic protein Bcl-2 decreased gradually.CONCLUSION: Ginsenoside Rh4 induces apoptosis of human hepatocellular carcinoma HepG2 cells, and the main mechanism may be related to down-regulation of Bcl-2 and up-regulation of Bax, cleaved caspase-3, and caspase-9.

Mini clinical evaluation exercise ( mini-CEX ) is an educational and assessable tool to evaluate the clinical skills among resident doctors .An examiner directly examines an examinee and the evaluate the process including history collection , physical examination , and the communication skills with the patients , diagnosis and treatment .This evaluation method is widely used for its convenience , time saving, and high availability.It is also applied to student nurses assessment but arises some problems during the practice at present.The main problems include inappropriate application without items adjustment , absence of the marking criterion establishment , and the non-standard teacher training system .This paper is mainly focus on the analysis of the existing problems , in order to provide references for further investigation and application of mini-CEX in the nursing education field .

Objective To investigate the alterations and phenotypes of dendritic cells, inflamma-tory monocytes and macrophages in immunocompetent mice during Pneumocystis murina ( P.murina) infec-tion for further analysis of the function of these cells during P.murina infection.Methods Wild type male C57BL/6 mice at age 6-8 weeks were randomly divided into two groups including the group with P.murina infection and the group receiving sham surgery.The mice without any intervention were used to set up the blank control group.The loads of P.murina strains in lung tissues of each mouse were quantified by TaqMan real-time fluorescence polymerase chain reaction after the infection.Histopathological examination was per-formed to evaluate the degree of inflammation in lung tissues.The numbers of dendritic cells, inflammatory monocytes and macrophages in lung tissues, peripheral blood and bone marrow samples, and the changes of inflammatory monocytes in spleen tissues were measured by flow cytometry analysis.The expression of major histocompatability complexⅡ(MHCⅡ), CX3C chemokine receptor 1 (CX3CR1) and CC chemokine re-ceptor 2 ( CCR2 ) by dendritic cells, inflammatory monocytes and macrophages in lung tissues during P.murina infection were analyzed by flow cytometry analysis.All of the data were collected one, two, three and four weeks after the corresponding treatments.Results The loads of P.murina strains in P.murina in-fected mice were elevated after two and three weeks infection, but decline at week 4 (P>0.05).Significant pathological changes including the alveolar destruction, inflammatory cell infiltration and thickened alveolar septum in mice with P.murina infection were observed under a microscope at week 3 and week 4.Compared to the sham surgery treatment group, the number of CD11c+CD11b+dendritic cells were increased in lung tissues, but decreased in blood samples during P.murina infection ( P<0.05) .The levels of inflammatory monocytes in blood samples fell at week 3 and then rose at week 4 during P.murina infection (P<0.05). No significant difference with the change of macrophages in mice was observed during P.murina infection ( P>0.05).The CD11c+CD11b+dendritic cells in lung tissues of mice with P.murina infection expressed high levels of MHCⅡand CX3CR1, and low levels of CCR2.The inflammatory monocytes in lung tissues of mice expressed high levels of CCR2, moderate levels of MHCⅡand low levels of CX3CR1 during P.murina in-fection.High levels of CX3CR1 and low levels of MHCⅡ and CCR2 were observed in macrophages from lung tissues of mice with P.murina infection.Conclusion Highly expressed CD11c+CD11b+dendritic cells and MHCⅡwere detected in lung tissues of mice during P.murina infection, indicating that CD11c+CD11b+dendritic cells were involved in the host defense against P.murina infection.

To improve the vocational nursing students' interest in learning physiology,‘four modernizations' teaching method was preliminarily explored in physiology teaching practice.‘Four modernizations' referred to the teaching by using multimedia.Using this method,we can simplify complex issues,inspire students' curiosity and thirst for knowledge,combine with real life,make abstract problem specific,mobilize students' initiative,contact with clinical practice,visualize theoretical issues,strengthen the importance and understanding and make moderate expansion so that professional skills and practicality of nursing can be emphasized.