Three years of COVID-19 pandemic: comparative analysis of incidence, lethality and mortality among the States of the south region of Brazil / Três anos de pandemia da COVID-19: análise comparativa da incidência, letalidade e mortalidade entre os Estados da região sul do Brasi

Introduction: the first COVID-19 case in Brazil was confirmed on February 26, 2020. As of March 17, 2023, the Ministry of Health reported 699,634 deaths from COVID-19, with a case fatality rate of 1.9%. The impact of the COVID-19 pandemic in Brazil extends to socioeconomic and healthcare systems, reflecting significant regional disparities. Objective: To analyze mortality, incidence, and case fatality rates for COVID-19 in the states of Paraná and Santa Catarina, in the southern region of Brazil. Methods: This is an ecological time-series study using official Brazilian secondary data for COVID-19 cases and deaths. Data were extracted from the dashboard of the State Health Department of Santa Catarina and Paraná. Temporal series were developed for trend analysis using the Prais-Winsten regression model. Statistical analyses were performed using STATA 14.0 software (College Station, TX, USA, 2013). Results: In the analysis of rates over the entire period, trends for mortality, case fatality, and incidence in the state of Santa Catarina are decreasing, decreasing, and stationary, respectively. In Paraná, rates over the entire period showed a stationary trend for mortality, decreasing for case fatality, and increasing for incidence. Conclusion: COVID-19 had a devastating effect on the states of Santa Catarina and Paraná. Both states experienced the progression of the COVID-19 pandemic, with higher case fatality and mortality rates observed in Paraná, while Santa Catarina had a higher incidence rate over the three years of the COVID-19 pandemic.

Clinical study of acupuncture combined with medication in the treatment of skin pruritus in maintenance hemodialysis patients / 针灸推拿医学（英文版）

Objective:To observe the clinical effect of acupuncture combined with Western medicine in the treatment of skin pruritus in maintenance hemodialysis patients.Methods:Eighty patients were randomly divided into a control group and an observation group,with 40 cases in each group.The control group was given loratadine orally,and the observation group was given acupuncture treatment in addition to the treatment used in the control group.The four-item itch questionnaire(FIIQ)score,indicators for skin barrier function,and serum interleukin(IL)-2 and IL-31 levels were compared.The efficacy was judged after the treatment ended.Results:The total effective rate was higher in the observation group than in the control group(P<0.05).After treatment,the site,frequency,severity of pruritus,sleep impact sub-scores,and FIIQ total score in both groups were reduced compared with those before treatment(P<0.05),and all scores in the observation group were lower than those in the control group(P<0.05).The stratum corneum hydration(SCH)and transepidermal water loss(TEWL)in the V-shaped area of the chest,the flexor side of the forearm,and the extensor side of the lower leg were not significantly changed in the control group(P>0.05);the SCH and TEWL in the V-shaped area of the chest,the flexor side of the forearm,and the extensor side of the lower leg in the observation group were improved(P<0.05),and all were better than those in the control group(P<0.05).The serum IL-2 and IL-31 levels in the control group did not change significantly(P>0.05);the serum IL-2 and IL-31 levels in the observation group were both significantly decreased(P<0.05)and were lower than those in the control group(P<0.05).Conclusion:Acupuncture combined with loratadine is highly effective in the treatment of pruritus in maintenance hemodialysis patients,and it can relieve pruritus,improve skin barrier function,and reduce serum IL-2 and IL-31 levels.

Expression of interleukin-31 in hypertrophic scar from postoperative incision tissue / 中华医学美学美容杂志

Objective:To explore the interleukin-31 protein expression in the hypertrophic scar of incision tissue after surgery and its underlying pathological impact.Methods:From February 2022 to February 2023, three HS patients scar tissue (HS) and their normal skin tissue (Control, NS) were obtained. Two patients were female and one patient was male. The tissues were fixed in 4% formalin and embedded in paraffin. Haematoxylin-eosin (HE) stain and immunohistochemical stain were used to evaluate the epidermal thickness, myofibroblasts of dermis and the expression level of IL-31 between HS and NS.Results:The epidermis thickness was (303.88±46.03) μm in HS group, while (133.02±17.40) μm in NS group ( t=12.60, P<0.001). The expression level of IL-31 protein was measured by IRS score and positive cell density. The IRS score was 9.89±2.03 of the basal layer in HS group and was 4.33±1.66 of the basal layer in NS group. The positive cell density was 786 343.83±159 627.97 of the basal layer in HS group ( P<0.001) and was 555 457.61±128 097.21 of the basal layer in NS group ( P=0.014). In the dermis layer, the IRS score was 7.11±1.05 in HS group and was 4.33±0.71 in NS group, the positive cell density was 156 760.97±26 046.10 in HS group ( P<0.001) and was 49 576.01±52 369.33 in NS group ( P<0.001). In the dermis layer, the count of myofibroblasts was 120.44±15.75 in HS group while was 27.39±14.89 in NS group ( t=23.79, P<0.001). Conclusions:Our study demonstrates that both myofibroblast count and IL-31 protein expression level are notably increased in HS patients. The expression of IL-31 protein is prominent in the cytoplasm of myofibroblasts, basal cells, macrophages and mast cells which could implicate that IL-31 may be a potential therapeutic target to enhance the resolution of HS.

Effects of circular RNA-SEC31A on the invasion and migration of pancreatic cancer cells and molecular mechanism / 中华胰腺病杂志

Objective:To determine the expression of circular RNA-SEC31A(circSEC31A) in pancreatic cancer and investigate the effects on the invasion and migration of pancreatic cancer cells and the underlying molecular mechanism.Methods:Differentially expressed circRNAs between pancreatic cancer cells (BXPC-3, PANC1, CaPan-2, SW1990) and human normal pancreatic cells (HPDE) were identified by qRT-PCR. Then, two cell lines with high circSEC31A expression were selected to conduct next experiments. According to the sequence of the back-splicing site in circSEC31A, siRNAs for downregulation of circSEC31A were designed and transfected by liposome to silence circSEC31A in pancreatic cancer cells, and grouped as followed siR-circSEC31A#1 and siR-circSEC31A#2. Meanwhile, siR-NC group transfected with non-specific siRNA served as control. Transwell assays and wound healing assays were operated to evaluate the functional role of circSEC31A on the invasion and migration of pancreatic cancer cells. RNA Pull-down assay with circSEC31A probe and oligo control probe was used to screen the miRNA combining with circSEC31A and the effects of miRNA on cell invasion and migration of pancreatic cancer cells were validated. The effect of miR-200c-3p and circSEC31A silencing on the expression of PDK1 mRNA was identified by qRT-PCR. The protein expression of PDK1, downstream Akt and p-Akt after circSEC31A silencing was verified by Western blotting assays.Results:The relative expression level of circSEC31A in HPDE (1.000±0.120) was obviously lower than that in BXPC-3 (1.920±0.130), SW1990 (2.93±0.528), PANC1 (4.557±0.692) and CaPan-2 (5.247±0.194), and all the differences were statistically significant ( P<0.001). Compared with the PANC1 siR-NC group (1301.3±94.6) and CaPan-2 siR-NC group (1835.0±70.1) per 100 high power field, transwell assays showed that the numbers of invasive pancreatic cancer cells was highly decreased in PANC1 siR-circSEC31A#1 group (727.3±92.9), siR-circSEC31A#2 group (792.0±18.1), CaPan-2 siR-circSEC31A#1 group (718.0±90.6), siR-circSEC31A#2 group (692.7±84.8). Wound healing assays showed that silencing circSEC31A decreased the wound healing rate of pancreatic cancer cells in PANC1 siR-circSEC31A#1 group (20.667±3.215)%, siR-circSEC31A#2 group (20.000±4.583)%, CaPan-2 siR-circSEC31A#1 group (28.000±8.185)%, siR-circSEC31A#2 group (29.667±5.686)%, compared with the PANC1 siR-NC group (55.000±4.359)% and CaPan-2 siR-NC group (69.000±3.606)%. RNA Pull-down assays showed that compared with PANC1 oligo probe group (1.000±0.091) and CaPan-2 oligo probe group (1.000±0.153), miR-200c-3p was significantly enriched in the PANC1 circSEC31A probe group (2.237±0.175) and CaPan-2 circSEC31A probe group (2.166±0.156). Compared with PANC1 siR-NC group (939.3±57.0) and CaPan-2 siR-NC group (786.7±51.5) per 100 high power field, the numbers of invasive pancreatic cancer cells were up-regulated in PANC1 siR-miR-200c-3p group (1206.0±99.1) and CaPan-2 siR-miR-200c-3p group (1838.0±105.7), while the low numbers of invasive pancreatic cancer cells were observed in PANC1 siR-miR-200c-3p+ siR-circSEC31A group (932.7±116.4) and CaPan-2 siR-miR-200c-3p+ siR-circSEC31A group (785.3±58.8). Compared with PANC1 siR-NC group (1.000±0.103) and CaPan-2 siR-NC group (1.000±0.107), the relative expression of PDK1 mRNA in PANC1 siR-miR-200c-3p group (1.898±0.159) and CaPan-2 siR-miR-200c-3p group (2.102±0.337) was upregulated. Furthermore, the expression of PDK1 mRNA was decreased in the siR-miR-200c-3p+ siR-circSEC31A group (0.980±0.070, 1.015±0.079). Western blot assays showed that the expression of PDK1 protein in PANC1 siR-NC group, siR-circSEC31A#1 group, siR-circSEC31A#2 group was 0.767±0.086, 0.281±0.191, 0.333±0.062 and in CaPan-2 siR-NC group, siR-circSEC31A#1 group, siR-circSEC31A#2 group was 0.712±0.038, 0.353±0.061, 0.308±0.018. The expression of p-Akt protein in PANC1 siR-NC group and siR-circSEC31A group was 0.741±0.050, 0.114±0.027, 0.139±0.041. In addition, p-Akt protein expression in CaPan-2 siR-NC group and siR-circSEC31A group was 0.823±0.052, 0.141±0.045, 0.280±0.089. PDK1 and p Akt expression in siR circSEC31A group was obviously lower than those in sir NC group. All the differences between either groups above were statistically significant ( P<0.05). Conclusions:circSEC31A is upregulated in pancreatic cancer cells, which facilitates the invasion and metastasis of pancreatic cancer cells via miR-200c-3p/PDK1/Akt signaling pathway, supporting that circSEC31A may function as a new diagnostic and therapeutic target for pancreatic cancer patients.

MiR⁃31 Promotes the Proliferation and Migration of Human Dental Pulp Mesenchymal Stem Cells / 中国生物化学与分子生物学报

Pulpitis and periapical inflammation are two common diseases in stomatology today. Existing treatment options primarily include root canal therapy and pulp revascularization, which can effectively control inflammation and preserve the affected tooth while also causing permanent deactivation of the pulp tissue, structural failure, and secondary infection. In recent years, research on dental pulp regeneration has progressively entered the public consciousness because of tissue engineering technology that combines stem cells and biomaterials. Due to their multi⁃differentiation and high proliferation, dental pulp stem cells (DPSCs) isolated from permanent or deciduous teeth have emerged as a significant stem cell source for dentin or pulp tissue regeneration. However, the number and survival time of live cells in the dam⁃ aged area are impacted, which significantly limits the efficacy of stem cells since they are unable to efficiently be recruited to the injured area. The ability of DPSCs to migrate and multiply must therefore be enhanced. This study sought to determine if miR⁃31 (miR⁃31) may significantly enhance the proliferative and migratory capacities of DPSCs. The tissue block enzyme digestion method was used to successfully separate and culture DPSCs from dental pulp tissues, and the miR⁃31 levels in dental pulp tissues and DPSCs from normal and inflammatory teeth were compared. The results of real⁃time fluorescence quanti⁃ tative PCR (RT⁃qPCR) revealed that the expression level of miR⁃31 in dental pulp tissues and DPSCs from inflammatory teeth was significantly lower when compared to the control group (P<0. 05). Interfer⁃ ence and over⁃expression of miR⁃31 expressions in DPSCs were specifically divided into three groups: the NC group, the miR⁃31 agomir (over⁃expressed) group and the miR⁃31 antagomir (inhibitor) group. RTq⁃PCR results showed that the transfection was successful (P<0. 001). The results of CCK⁃8, wound⁃ healing, and Transwell migration experiments showed that overexpression of miR⁃31 successfully improved the proliferation and migration abilities of DPSCs compared with the control group (P<0. 05). Further⁃ more, Western blotting analysis revealed that miR⁃31 overexpression increased the expression of important migratory proteins, including CXC chemokine receptor type 4 (CXCR4) and matrix metalloproteinase2 (MMP2), as well as key proliferation proteins Ki67 and proliferating cell nuclear antigen (PCNA) (P< 0. 05). This study demonstrates that miR⁃31 can effectively boost the proliferation and migratory ability of DPSCs, providing strong theoretical support for the increased use of DPSCs in regenerative medicine.

Construction of annexin A2 plasmid and its role in the proliferation and migration of bladder cancer pumc-91 cells / 西安交通大学学报(医学版)

【Objective】 Annexin A2 (annexin A2, Anxa2) has been reported to regulate bioactivity in various tumors cells. The purpose of this study was to investigate the correlation between the expression of Anxa2 protein and the proliferation and migration abilities of bladder cancer pumc-91 cells. 【Methods】 The ANXA2 sequence was amplified and inserted into the pcDNA3.1(+) vector in order to prepare the pcDNA3.1(+)-ANXA2 plasmid. PcDNA3.1 (+)-ANXA2 was transiently transfected into pumc-91 bladder cancer cells by lipofectamine 2000. Western blotting assay was performed to detect the expression of Anxa2 protein in the blank group, the control group transfected with pcDNA3.1(+), and the experimental group transfected with pcDNA3.1(+)-ANXA2 plasmid. The proliferation ability of pumc-91 cells was detected using Cell Counting Kit-8(CCK8), and the migration level of pumc-91 cells was detected by transwell assay. Differences in detection data among the groups were compared using one-way ANOVA or repeated measures ANOVA. 【Results】 The plasmid construction was successful and the sequencing was absolutely correct. Western blotting assay showed elevated Anxa2 protein expression level in the experimental group compared to the blank and control groups. CCK8 assay suggested that the number of proliferating pumc-91 cells was significantly higher in the experimental group than in the blank group (P<0.001) and the control group (P=0.001). Transwell assay also showed that the number of pumc-91 cells crossing the membrane was significantly higher in the experimental group than in the blank group (P=0.011) and the control group (P=0.027). 【Conclusion】 Our findings suggested that up-expression of Anxa2 may play a critical role in regulating proliferation and migration of bladder cancer pumc-91 cells.

Investigation on "Fengshi (GB 31) for treating wind disorders" / 中国针灸

From the perspective of academic history, the paper reviews systematically the background and evolution of the understanding of "Fengshi (GB 31) for treating wind disorders". In the ancient literature, there are no direct relevant statement for the indication of Fengshi (GB 31) associated with "wind", and the consensus on "Fengshi for treating wind disorders" has not been made yet. Under the influence of acupoint theory in recent era and the syndrome differentiation for acupuncture treatment in modern time, this statement becomes a conventional understanding and acceptable gradually. Meanwhile, the understanding for Fengshi (GB 31) treating wind disorders tends to be generalized. Practically, Fengshi (GB 31) is applicable for the various disorders in the local and adjacent areas. It is necessary for modern acupuncture researchers to systematically collate, investigate and identify the knowledge content with a sense of familiarity so that the contemporary inheritance, development and application of traditional theoretical knowledge of acupuncture can be enhanced.

Mechanism of regulatory effect of recombinant human HMGB1 on endothelial cell angiogenesis / 器官移植

Objective To unravel the possible mechanism of the role of recombinant human high mobility group box 1 (rhHMGB1) protein in regulating the angiogenesis of endothelial cells. Methods Endothelial cells were divided into the control group, bone marrow mesenchymal stem cells (MSC) supernatant group and rhHMGB1 group. The proliferation and survival of endothelial cells were detected by cell counting kit(CCK)-8 assay. The relative expression levels of vascular endothelial growth factor (VEGF), Yes-associated protein (YAP), CD31 and hypoxia inducible factor (HIF)-1α proteins were determined by Western blot. The relative expression levels of VEGF, YAP, CD31 and HIF-1α messenger RNA (mRNA) were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The migration ability of endothelial cells was assessed by Transwell chamber test. The localization of YAP was detected by immunofluorescence staining. Results Compared with the control group, the migration rate of endothelial cells was increased in the rhHMGB1 group (P < 0.05), and the cell migration rate was enhanced over time. Compared with the control group, the relative expression levels of VEGF and p-YAP proteins were up-regulated in the MSC supernatant group, and the differences were statistically significant (both P < 0.05). Compared with the control group, the relative expression levels of VEGF and HIF-1α proteins, VEGF and CD31 mRNA and YAP and p-YAP proteins were up-regulated, and YAP/p-YAP ratio was increased in the rhHMGB1 group, and the differences were statistically significant (all P < 0.05). Compared with the MSC supernatant group, the relative expression levels of CD31 mRNA and YAP protein were up-regulated, and the YAP/p-YAP ratio was increased in the rhHMGB1 group, and the differences were statistically significant (all P < 0.05). Conclusions Exogenous high-concentration rhHMGB1 may promote the migration ability of endothelial cells and up-regulate the expression levels of angiogenesis-related proteins by regulating the recruitment of YAP to the nucleus.

Soluble epoxide hydrolase inhibitor promotes the healing of oral ulcers

Abstract Objective Oral ulcers are a lesion in the oral mucosa that impacts chewing or drinking. Epoxyeicosatrienoic Acids (EETs) have enhanced angiogenic, regenerative, anti-inflammatory, and analgesic effects. The present study aims to evaluate the effects of 1-Trifluoromethoxyphenyl-3-(1-Propionylpiperidin-4-yl) Urea (TPPU), a soluble epoxide hydrolase inhibitor for increasing EETs level, on the healing of oral ulcers. Methods The chemically-induced oral ulcers were established in Sprague Dawley rats. The ulcer area was treated with TPPU to evaluate the healing time and pain threshold of ulcers. The expression of angiogenesis and cell proliferation-related protein in the ulcer area was detected using immunohistochemical staining. The effects of TPPU on migration and angiogenesis capability were measured with scratch assay and tube formation. Results Compared with the control group, TPPU promoted wound healing of oral ulcers with a shorter healing time, and raised pain thresholds. Immunohistochemical staining showed that TPPU increased the expression of angiogenesis and cell proliferation-related protein with reduced inflammatory cell infiltration in the ulcer area. TPPU enhanced cell migration and tube-forming potential in vitro. Conclusions The present results support the potential of TPPU with multiple biological effects for the treatment of oral ulcers by targeting soluble epoxide hydrolase.

Cytotoxicity evaluation of haloperidol, clozapine and a new molecule with antipsychotic potential, PT-31, in NIH-3T3 cells

Abstract Schizophrenia is an illness that affects 26 million people worldwide. However, conventional antipsychotics present side effects and toxicity, highlighting the need for new antipsychotics. We aimed to evaluate the cytotoxicity of haloperidol (HAL), clozapine (CLO), and a new molecule with antipsychotic potential, PT-31, in NIH-3T3 cells. The neutral red uptake assay and the MTT assay were performed to evaluate cell viability and mitochondrial activity, morphological changes were assessed, and intracellular reactive oxygen species (ROS) detection was performed. HAL and CLO (0.1 µM) showed a decrease in cell viability in the neutral red uptake assay and in the MTT assay. In addition, cell detachment, content decrease, rounding and cell death were also observed at 0.1 µM for both antipsychotics. An increase in ROS was observed for HAL (0.001, 0.01 and 1 µM) and CLO (0.01 and 1 µM). PT-31 did not alter cell viability in any of the assays, although it increased ROS at 0.01 and 1 µM. HAL and CLO present cytotoxicity at 0.1 µM, possibly through apoptosis and necrosis. In contrast, PT-31 does not present cytotoxicity to NIH-3T3 cells. Further studies must be performed for a better understanding of these mechanisms and the potential risk of conventional antipsychotics

Is Atrophic Nonunion a Misnomer - A Hospital-based Prospective Cross-Sectional Study / Pseudoartrose atrófica é um termo incorreto? - Um estudo hospitalar transversal prospectivo

Abstract Objective The present study was conducted to estimate histologically the proportion of avascularity of fracture ends in case of nonunion of long bones. Methods A total of 15 cases of established quiescent nonunion were operated according to the standard protocol and the fracture ends were evaluated histologically. The biopsied tissue was briefly fixed with formalin, embedded with paraffin (FFPE), and 5-micron sections were stained with hematoxylin and eosin according to standard protocols. Immunohistochemistry with anti-CD31 antibody (JC70A clone, DBS) was performed manually using standard protocols. Results All cases of quiescent nonunion were included; radiologically, 2 cases were oligotrophic, and 13 cases were of atrophic nonunion. A total of 20% of the patients were females, 40% were in the age group between 31and 40 years old, and, radiologically, all cases were of atrophic nonunion. All cases showed positivity for CD-31 on immunohistochemistry. The blood vessel density was category I in 13.33% of the cases and category II in 86.67% of the cases. Four cases presented with mild inflammation and two presented with moderate inflammation. The average vessel count was 10 per high power field in the age groups between 20 and 30, 31 and 40, and 41and 50 years old. The age group between 61 and 70 years old showed an average vessel count of 4 per high power field. The difference in the vessel counts of oligotrophic and atrophic nonunion was not significant. No correlation was observed in the density of vessel count and duration of nonunion Conclusion The nomenclature for the classification of nonunion into atrophic, oligotrophic, and hypertrophic needs revision. Our findings do not support that atrophic and oligotrophic nonunion are histologically different.

Resumo Objetivo O presente estudo estimou a proporção de avascularidade histológica das extremidades das fraturas em caso de pseudoartrose de ossos longos. Métodos No total, 15 casos de pseudoartrose quiescente estabelecida foram operados de acordo com o protocolo padrão e as extremidades da fratura foram avaliadas histologicamente. Em resumo, o tecido biopsiado foi fixado em formalina e embebido em parafina (FFPE); secções de 5 mícrons foram coradas com hematoxilina e eosina de acordo com os protocolos padrões. A imunohistoquímica com anticorpo anti-CD31 (clone JC70A, DBS) foi realizada manualmente segundo protocolos padrões. Resultados Todos os casos de pseudoartrose quiescente foram incluídos; 2 eram de pseudoartrose oligotrófica e 13 eram de pseudoartrose atrófica à radiologia. Destes, 20% eram de pacientes do sexo feminino, 40% de indivíduos entre 31 e 40 anos de idade e todos os casos eram de pseudoartrose atrófica à radiologia. Todos os casos eram positivos para CD-31 à imunohistoquímica. A densidade dos vasos sanguíneos era de categoria I em 13,33% dos casos e de categoria II em 86,67%. Quatro casos apresentavam inflamação branda e dois apresentavam inflamação moderada. O número médio de vasos era de 10 por campo de alta potência na faixa etária de 20 a 30, de 31 a 40 e de 41 a 50 anos. A faixa etária de 61 a 70 anos apresentava, em média, 4 vasos por campo de alta potência. A diferença nos números de vasos em pseudoarthroses oligotróficas e atróficas não foi significativa. Não houve correlação entre a densidade de vasos e a duração da pseudoartrose. Conclusão A nomenclatura de classificação da pseudoartrose em atrófica, oligotrófica e hipertrófica precisa ser revista. Nossos achados não indicam que a pseudoartrose atrófica e oligotrófica sejam histologicamente diferentes.

Mechanisms of pruritus in bullous pemphigoid / 中华皮肤科杂志

Pruritus is one of the typical clinical manifestations of bullous pemphigoid (BP) . In recent years, researchers have gradually recognized that the histamine-independent itch pathway plays an important role in BP. Eosinophils, basophils, interleukin (IL) -31, IL-4, IL-13, thymic stromal lymphopoietin, periostin and substance P are all closely related to the occurrence of pruritus in BP. This review mainly elaborates research progress in mechanisms related to pruritus in BP.

Correlation of peripheral blood miR-31 and oxidative stress indicators of diabetic nephropathy / 中国医师杂志

Objective:To explore and analyze the correlation between miR-31 in peripheral blood and oxidative stress indicators of diabetic nephropathy.Methods:A total of 94 patients with diabetic nephropathy who were admitted to Affiliated Hospital of Jining Medical College from September 2019 to September 2020 were selected. Patients were divided into mild diabetic nephropathy [estimated glomerular filtration rate (eGFR) 60-90 ml/min, 36 cases] group, moderate diabetic nephropathy (eGFR 30-60 ml/min, 27 cases) group and severe diabetic nephropathy (eGFR 0-30 ml/min, 31 cases) group according to the severity of the disease, and 30 healthy people in the same period were selected as the control group. Real time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-31 in peripheral blood. Serum superoxide dismutase (SOD), malondialdehyde (MDA), advanced oxidation protein products (AOPP) and other oxidative stress indicators, as well as serum urea nitrogen, creatinine and glomerular filtration rate. Pearson was used to analyze the correlation between peripheral blood miR-31 and oxidative stress indexes and renal function.Results:The expression of miR-31 in peripheral blood of patients with diabetic nephropathy was significantly lower than that in the control group, and the expression of miR-31 in peripheral blood of patients in severe and moderate diabetic nephropathy group was significantly lower than that in the mild diabetic nephropathy group (all P<0.05), with statistically significant difference (all P<0.05). Pearson correlation analysis showed that serum miR-31 expression was negatively correlated with the severity of diabetic nephropathy ( r=-0.526, P<0.05). The levels of serum MDA, SOD and AOPP in the diabetic nephropathy group were significantly higher than those in the control group, and the levels of serum MDA, SOD and AOPP in the severe and moderate diabetic nephropathy groups were higher than those in the mild diabetic nephropathy group, with statistically significant difference (all P<0.05). The levels of serum creatinine and blood urea nitrogen in the diabetic nephropathy group were higher than those in the control group, and the glomerular filtration rate was lower than that in the control group (all P<0.05). The levels of serum creatinine and blood urea nitrogen in the severe and moderate diabetic nephropathy group were higher than those in the mild diabetic nephropathy group, while the level of glomerular filtration rate was lower than that in the mild diabetic nephropathy group, with statistically significant difference (all P<0.05). Pearson correlation analysis showed that the expression of miR-31 in peripheral blood was negatively correlated with the levels of MDA, SOD, AOPP, serum creatinine and urea nitrogen (all P<0.05), but positively correlated with glomerular filtration rate ( P<0.05). Conclusions:The expression of miR-31 in peripheral blood gradually decreases with the severity of renal damage. Its level is negatively correlated with oxidative stress indicators of diabetic nephropathy, and positively correlated with glomerular filtration rate, which can be used for for clinical treatment and disease evaluation.

Immunoregulatory effects of miR-31-5p overexpression on peripheral blood Th17 cell in a rabbit model of autoimmune dry eye / 中华实验眼科杂志

Objective:To investigate the immunoregulatory effects of lentivirus-mediated microRNA (miR)-31-5p overexpression on peripheral blood T helper cell 17 (Th17) in a rabbit model of autoimmune dry eye.Methods:The miR-31-5p recombinant lentiviral vector was constructed.Lentivirus overexpressing miR-31-5p and its control virus were packaged.The concentration measurement and lentiviral titer determination were carried out.A rabbit model of autoimmune dry eye was established and the peripheral blood mononuclear cells (PBMC) of the rabbits were isolated.PBMC infected with miR-31-5p and negative control lentivirus particles were assigned as the miR-31-5p overexpression group and control group, respectively.The miR-31-5p expression level was detected using quantitative real-time PCR (qRT-PCR). Then PBMC in the two groups were co-cultured with γ-ray irradiated lacrimal gland epithelial cells.The expressions of Th17 cell related transcription factor retinoic acid-receptor-related orphan receptor C (RORC) and interleukin-17 (IL-17) mRNA, IL-1β, IL-6 and IL-23 were determined by qRT-PCR.The IL-17 protein expression level was detected by Western blot.The use and care of animals complied with Regulation for the Administration of Affair Concerning Experiment Animals by State Science and Technology Commission.The study protocol was approved by the Ethics Committee of Tianjin Medical University Eye Hospital (No.TJYY20201221036).Results:The construction of the miR-31-5p recombinant lentiviral vector was verified by DNA sequencing.The lentiviral titer of lentivirus overexpressing miR-31-5p and control lentivirus particles was 3.82×10 7 TU/ml and 3.50×10 7 TU/ml, respectively.The miR-31-5p relative expression level of PBMC was significantly increased in miR-31-5p overexpression group in comparison with control group, showing a statistically significant difference ( t=-9.696, P<0.001). When PBMC were co-cultured with lacrimal gland epithelial cells in vitro, the relative expression levels of RORC and IL-17 mRNA in miR-31-5p overexpression group were 0.33±0.03 and 0.28±0.09, which were significantly decreased in comparison with 1.00±0.00 and 1.00±0.00 in control group, with statistically significant differences between them ( t=46.256, 13.810; both at P<0.05). The relative expression level of IL-17 protein in miR-31-5p overexpression group was significantly reduced than control group ( t=4.977, P=0.008). The relative expression levels of IL-1β, IL-6 and IL-23 mRNA were significantly lower in miR-31-5p overexpression group than control group ( t=220.076, 6.641, 13.271; all at P<0.05). Conclusions:The overexpression of miR-31-5p can inhibit the Th17-immune response via down-regulating the expression of IL-6, IL-1β and IL-23.

Ameliorative Effect of miR-31 on Liver Injury in Type 2 Diabetic Mice / 中国生物化学与分子生物学报

Type 2 diabetes mellitus (T2DM) is a metabolic disease with an increasing incidence worldwide, which leads to damage to various tissues and organs including the liver. MiR-31 is conserved across species and closely associated with metabolic diseases, but its role in type 2 diabetic liver injury has not been elucidated. This study aimed to investigate the effect of miR-31 on liver injury in type 2 diabetes and its underlying mechanism. Four to six weeks old male FVB mice and miR-31-positive transgenic mice were randomly divided into FVB mice control group (C), FVB mice induced diabetes group (DM) and miR-31-overexpression transgenic mice induced diabetes group (31DM). After 1 week of adaptive feeding, the T2DM mouse model was induced by high-fat feeding combined with intraperitoneal injection of streptozotocin (STZ) for 6 weeks. The general condition of mice and related metabolic indicators showed that the increased food and water intake, weight loss and glucose and lipid metabolism disorders could be reversed by miR-31 in T2DM mice. HE staining and liver histological activity index (HAI) scoring results showed that miR-31 improved the inflammatory status in the liver tissue of T2DM mice and decreased the HAI score. RT-qPCR results showed that the high expression of miR-31 was accompanied by a decrease in the expression of activating transcription factor 6 (ATF6) mRNA in the liver of T2DM mice. Furthermore, Western blotting results showed that miR-31 inhibited the expression of endoplasmic reticulum stress-related proteins such as ATF6, glucoregulatory protein 78 (GRP78) and C/EBP homologous protein (CHOP) in the liver of T2DM mice. In conclusion, miR-31 may ameliorate liver injury in T2DM mice by regulating glucose and lipid metabolism disorders and insulin resistance, and inhibiting endoplasmic reticulum stress factors such as ATF6, GRP78, and CHOP.

Inhibitory Effect of Duanteng Yimu Decoction on Pannus Formation in Mice with Collagen-induced Arthritis Based on VEGF Pathway / 中国实验方剂学杂志

Objective:To investigate the mechanism of Duanteng Yimu decoction （DTYM） in the inhibition of pannus formation in collagen-induced arthritis （CIA） mice. Method:Twenty-four SPF-grade DBA/1 male mice were randomly divided into the following four groups： a blank group （NC group）， a model group （CIA group）， a methotrexate group （MTX group）， and a DTYM group， with six mice in each group. The mice， except for those in the NC group， were modeled. From the second immunization， the medium， MTX （1 mg·kg^-1）， and DTYM （15.4 g·kg^-1） were administered at an equal volume by gavage for 35 days. Mice were observed for general condition and the arthritis index. The knee and ankle joints were scanned by microcomputed tomography （micro CT）. Hematoxylin-eosin （HE） and safranin O/fast green staining were performed to observe pathological changes. Immunohistochemistry was performed to detect the expression of platelet/endothelial cell adhesion molecule-1 （CD31）， vascular endothelial growth factor-<italic>α</italic> （VEGF-<italic>α</italic>）， vascular endothelial growth factor receptor 2 （VEGFR2）， and phosphorylated（p）-VEGFR2. Result:Compared with the NC group， the CIA group showed red and swollen ankle joints， increased arthritis index scores （<italic>P</italic><0.05， <italic>P</italic><0.01）， manifest injury in the knee and ankle joints， reduced cartilage thickness， elevated Micro CT bone destruction scores of knee and ankle joints （<italic>P</italic><0.01）， and up-regulated absorbance values of synovial CD31， VEGF-<italic>α</italic>， VEGFR2， and p-VEGFR2 （<italic>P</italic><0.01）. Compared with the CIA group， the DTYM group showed relieved ankle joint redness and swelling， reduced arthritis index scores of mice three weeks after administration （<italic>P</italic><0.05， <italic>P</italic><0.01）， intact joint surfaces of the knee and ankle joints， thickened cartilage， declining Micro CT bone destruction scores in both the knee and ankle joints （<italic>P</italic><0.05， <italic>P</italic><0.01）， and lowered absorbance values of CD31， VEGF-<italic>α</italic>， VEGFR2， and p-VEGFR2 in the synovium （<italic>P</italic><0.01）. Conclusion:DTYM can inhibit the pannus formation in CIA mice presumedly by regulating the VEGF pathway.

Localization of Melanoma-associated Antigen MAGEA1 and Identification of Its Interacting Proteins / 中国生物化学与分子生物学报

Melanoma associated antigen family A1 (MAGEA1) is expressed in germ cells and tumors of various histological origins, but its mechanism is still unclear. In this study, the eukaryotic recombinant MAGEA1 expression plasmids with Flag or GFP tags were constructed and transfected into HeLa and HEK293T cells. Western blotting, immunocytochemistry, co-immunoprecipitation, nuclear protein and cytoplasmic protein separation, and mitochondrial isolation were used to detect the expression and location of MAGEA1 and its interaction with other proteins in cells. The results of immunocytochemistry (ICC) and Western blotting showed that the overexpressed MAGEA1 was mainly localized in the cytoplasm and partially co-localized with mitochondria. Co-immunoprecipitation experiments verified the interactions between MAGEA1 and TRIM31, SNW1, HDAC1, and found that MAGEA1 may mainly interact with HDAC1 in the cytoplasm. The studies above indicate that MAGEA1 may be involved in different cellular biological processes and co-localize with mitochondria. It interacts with TRIM31, SNW1 and HDAC1, while MAGEA1 may mainly interact with HDAC1 in the cytoplasm. We propose that it may be involved in protein ubiquitination and the Notch signaling pathway. The results of this study laid an experimental foundation for the subsequent in-depth study of the mechanism of MAGEA1.

Long non-coding RNA SPATA31D5P promoting proliferation, migration and invasion of breast cancer cells through adsorption of microRNA-320a / 解剖学报

Objective Long non-coding RNA(lncRNA) are aberrantly expressed in breast cancer(BC) and strongly associated with its survival prognosis. The aim of this study is to investigate the expression and effect of IncRNA SPATA31D5P on the invasion and migration capacity of breast cancer cells through adsorption of miR-320a. Methods Totally 30 cases of BC tissues and paraneoplastic tissues were collected, and the expression levels of SPATA31D5P in BC tissues and BC cell lines were detected by Real-time PCR. MDA-MB-231 cells were transfected with SPATA31D5P siRNA interference vector, and cell proliferation, invasion and migration capacity were determined using the cell counting kit-8 assay (CCK-8), 5-ethynyl-2'- deoxyuridine(EdU), Transwell and wound-healing assay respectively. And cell cycle and apoptosis were detected by flow cytometry. Bioinformatics approachs were used to screen for miRNAs that could bind complementarily to SPATA31D5P, and the regulatory effect of SPATA31D5P on miR-320a was detected by Real-time PCR and dual luciferase reporter assay. Results SPATA31D5P levels were significantly higher in BC tissues than in adjacent normal breast tissues, and SPATA31D5P expression was higher in each BC cell line than in normal breast epithelial cells MCF10 A. The level of SPATA31D5P in the interference group was 0. 288±0. 052, which was lower than that of the blank control group 1. 114±0. 096 and negative control (NC) group 1. 079±0. 128 (P< 0. 01). The proliferation activity of MDA- MB-231 cells in the interfered group was significantly reduced and apoptotic rate was obviously increased compared to the NC and control groups (P<0. 01) ;the Gj phase block was observed in the interfered group; the scratch healing rate and number of perforated cells in the interference group were (14. 36 ± 1. 75) % and (26±1.52), which were lower than (52. 25± 1.87)% and ( 67. 33 ± 2. 91 ) of the NC group (PcO.Ol). Dual luciferase experiments confirmed that SPATA31D5P could directly regulate miR-320a expression and luciferase activity. Conclusion SPATA31D5P is highly expressed in BC, interfering with SPATA31D5P expression effectively inhibits the proliferation, migration and invasion of MDA-MB-231 cells, and the mechanism may be related to the targeted regulation of miR-320a.

Protective effect of mitochondria-targeted antioxidant peptide SS-31 in sepsis-induced acute kidney injury / 中华危重病急救医学

Objective:To study the effect and mechanism of mitochondria-targeted antioxidant peptide SS-31 on sepsis-induced acute kidney injury (AKI).Methods:Sixty adult male C57BL/6 mice were randomly divided into four groups according to the random number table method: sham group (10 mice), positive control group (10 mice), sepsis model group (20 mice), and SS-31 peptide group (20 mice). The sepsis-induced AKI mouse model was reproduced by cecal ligation and puncture (CLP). The sham group only received laparotomy. SS-31 peptide (5 mg/kg) was intraperitoneally injected in SS-31 peptide group and positive control group 30 minutes after the operation, while an equivalent amount of normal saline was given in sham group and sepsis model group for 7 days. The blood samples were collected 24 hours after the operation from orbit, and the serum was collected to test the serum creatinine (SCr) and blood urea nitrogen (BUN). The mice were sacrificed 7 days after surgery. The kidney tissues were collected to observe the pathologic structure changes under the hematoxylin-eosin (HE) staining by light microscope. And the mitochondrial ultrastructure was checked under the transmission electron microscope. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method (TUNEL). The expression level of peroxisome proliferator-activated receptorγ coactivator-1α (PGC-1α), adenosine monophosphate-activated protein kinase (AMPK), and cleaved caspase-3 protein were tested by Western blotting.Results:Compared with sham group, the levels of SCr and BUN were significantly increased in sepsis model group [SCr (μmol/L): 93.12±11.80 vs. 32.94±3.37, BUN (mmol/L): 41.36±6.48 vs. 9.49±3.58, both P<0.05]. The expression levels of AMPK, PGC-1α and cleaved caspase-3 protein increased (AMPK/β-actin: 0.30±0.02 vs. 0.12±0.01, PGC-1α/β-actin: 0.38±0.03 vs. 0.16±0.02, cleaved caspase-3/β-actin: 0.20±0.01 vs. 0.11±0.02, all P<0.05). HE staining showed that inflammatory cell was infiltrated, glomerular basement membrane was exposed and vacuole-like transparent casts were found in the lumen. Mitochondria were damaged under electron microscope with swelling, ridge disappearance and ruptured membranes, with increasing of apoptotic cells [cells: 24.00 (18.75, 31.00) vs. 2.00 (0.72, 3.25) , P<0.05]. Meanwhile, compared with sepsis model group, the levels of SCr, BUN and the expressions of AMPK, PGC-1α, cleaved caspase-3 protein were significantly decreased in the SS-31 peptide group [SCr (μmol/L): 71.33±10.14 vs. 93.12±11.80, BUN (mmol/L): 27.00±5.52 vs. 41.36±6.48, AMPK/β-actin: 0.23±0.01 vs. 0.30±0.02, PGC-1α/β-actin: 0.27±0.02 vs. 0.38±0.03, cleaved caspase-3/β-actin: 0.13±0.01 vs. 0.20±0.01, all P < 0.05]. HE staining showed that cell swelling reduced, the mitochondrial structure was intact, the ridge swelling was also reduced, and the membrane structure was relatively intact, the number of apoptotic cells was significantly reduced [cells: 13.00 (9.00, 16.50) vs. 24.00 (18.75, 31.00) , P<0.05]. Conclusion:The protective effect of SS-31 peptide on organ dysfunction induced by sepsis-induced AKI is related to maintaining mitochondrial homeostasis and inhibiting cell apoptosis.

Bengalis - ‘Do Business’: A Descriptive Study on Acharya Prafulla Chandra Roy’s Entrepreneurship Development

The main objective of this original research work is to explore the various entrepreneurship activitiesexecuted by Acharya Prafulla Chandra Roy (APC), the father of Indian Chemistry. The study period hasbeen taken from 1950 to 2020 i.e. 7 decades to analyze the sustainable impact of entrepreneurial activitiesof APC. The most relevant and appropriate information has been used in the study by accessing publishedbooks, reports, articles, news papers, video tapes, audio tapes of different reputed private and governmentagencies and institutions. For visual representation, some relevant images have been incorporated in thestudy. The study finally reveals that APC tried to create an immense impact on the lives of Indiansspecially Bengalis to take the route of entrepreneurship as a career option to create jobs and not for takingjobs. To the best of his capabilities and high degree enthusiasm even at the age of 80 he tried to addressIndian youngsters for travelling to the route of innovation & entrepreneurship, setting up factories,building entrepreneurship eco system and taking India forward. In the study it has also been found that,APC was in favour of qualitative education and not at all quantitative education by rote and made asignificant and serious attempt to criticize higher education in the universities. The study also reveals thatwith the help of research and development (R&D), creativity and innovation; APC had established BengalChemicals and Pharmaceuticals Limited (BPCL), having registered office in Kolkata (previously knownas Calcutta) and with that India’s first pharmaceutical company concept originated. With BPCL, APCtried to deliver world class health and hygiene product portfolio as well as innovative drug and health caresolution with most affordable fees across India and made a significant and sustainable impact on Indianconsumers. Finally in the study it has been found that the country’s oldest pharmaceuticals company,Bengal Chemicals & Pharmaceuticals Ltd. (BCPL), set up by APC is abruptly in the spotlight due to thesudden demand for hydroxychloroquine (HCQ), the most sought-after drug in the treatment of Covid-19.