Objective To investigate the expression pattern of microRNA (miRNA) in the kidneys of unilateral ureteral obstruction (UUO) rats and to identify specific miRNA related to renal interstitial fibrosis (RIF).Methods Forty-eight male SD rats were divided into two groups:UUO group and sham-operated (Sham) group.Rats were sacrificed at 3,7 and 14 days after operation.Histologic changes were examined by Masson staining.Forty-eight selected miRNAs were examined by stem-loop real-time qPCR.Results At the 3rd day after operation,obstructed kidneys from operation rats showed mild edema in the interstitium and mononuclear cell infiltration.At the 7th day after operation,focal interstitial fibrosis was observed.At the 14th day after operation,fibrosis became more severe.The Sham kidneys showed no pathological changes.At the 3th day after operation,25 miRNAs were differentially expressed.At the 7th day after operation,24 miRNAs were aberrantly expressed,whereas 21 miRNAs were differentially expressed at the 14th day after operation (P＜0.05).Among these miRNAs,miR-132,miR-192,miR-194,miR-29c and miR-203 were consistently up-regulated or down-regulated in a time-dependent manner after operation.There were significantly correlations between the expression of five miRNAs and severity of tubulointerstitial injury (P＜0.05).Conclusions There are at least 20 miRNAs differentially expressed in the process of RIF induced by UUO.There are significantly correlations between the expression of miR-132,miR-192,miR-194,miR-29c and miR-203 and the severity of tubulointerstitial injury.They may be closely related to RIF.A further study is needed.

Objective To explore the effect of Xinkang Tablets on myocardial apoptosis index,collagen volume fraction and sarcoplasmic reticulum Ca2+-ATPase activity of rats with adriamycin-induced heart failure.Methods The chronic heart failure (CHF) SD rat model was established by intraperitoneal injection of doxorubicin.After successful modeling,the rats with CHF were randomly divided into 5 groups,namely model group,western medicine group,and low-,middle-and high-dose of Chinese medicine groups,10 rats in each group.The rats in the above groups were given intragastric administration of distilled water,22.5 μg/kg of Digoxin mixed suspension,9,18,36 g/kg of XinkangTablets,respectively,in the volume of 10 mL/kg of distilled water dilution,once a day,for 5 continuous weeks.Another the same batch of 10 SD rats were randomly allocated to the sham operation group,and were treated with intragastric administration of the same volume of distilled water.And then the apoptotic rate of myocardial cells was measured by TUNEL method,the collagen volume fraction (CVF) was measured after Masson staining,and the sarcoplasmic reticulum Ca2+-ATPase activity was determined by inorganic phosphate assay.Results Compared with the sham operation group,the apoptotic rate of myocardial cells and CVF in the model group were increased(P ＜ 0.01),indicating that the myocardial remodeling occurred in rats with CHF.Compared with the model Group,the apoptotic rate of western medicine group and three Chinese medicine groups was significantly decreased(P ＜ 0.01),suggesting that Digoxin and Xinkang Tablets can relieve apoptosis to certain extent.The CVF in Digoxin group and middle-and high-dose of Chinese medicine groups were lower than those in the model Group (P＜ 0.05 or P＜ 0.01),indicating that Digoxin and Xinkang Tablets can delay the myocardial fibrosis.Last but not least,the SERCA2a activities in the middle-and high-dose of Chinese medicine groups were higher than those in the model group (P ＜ 0.05 or P ＜ 0.01),suggesting that Xinkang Tablets may relieve myocardial remodeling and improve cardiac function through the regulation of SERCA2a activity.Conclusion Xinkang Tablets decrease the apoptotic rate and myocardial cell volume fraction probably through the regulation of SERCA2a activity,which may play a role in counteracting apoptosis and myocardial fibrosis,and ultimately delay the remodeling of the myocardium.

Objective To investigate the changes of A20 expression stimulated by free fatty acids (FFA) and its pathway.Methods HepG2 cells and U937 cells were stimulated by 0.5 mmol/L mixed FFA.The expression of A20,phosphor-p65 and phosphor-IκBα of neclear factor (NF)-κB pathway and phosphor-c-Jun N-terminal kinase (JNK),JNK,phosphor-extracellular signal-regulated kinase (ERK),ERK,phosphor-p38 and p38 of mitogen-activated protein kinase (MAPK) pathway were detected by Western blotting.The level of interleukin (IL)-12p,IL-1β,tumor necrosis factor (TNF)-α,IL-6,IL-10 and IL-8 cytokines in the supernatant of cell culture were detected by flow cytometry.T-test was performed for statistical analysis.Results The level of A20 changed along with the stimulated time of FFA.NF-κB and MAPK pathways were activated after FFA stimulation.The secretion of IL-6 and IL-8 increased after HepG2 cells stimulated by FFA and both reached peak at 24 hour.Compared with control group,the difference in IL-8 was statistically significant ((423.8 ± 8.9) pg/mL vs (12.4 ± 4.5) pg/mL,t=41.28,P＜0.01).The difference in IL-6 was also statistically significant ((4 082±423.6) pg/mL vs (52.9±29.5) pg/mL,t=9.49,P＜0.01).After U937 cells were stimulated by FFA,the secretion of IL-8 increased compared with control group.And in a certain period of time the secretion was time dependence.The maximum secretion of 24 hours was (200.6±5.7) pg/mL vs (5.0±3.9) pg/mL,and the difference was statistically significant (t=28.16,P＜0.01).IL-10,IL-12p,IL-1β and TNF-α were detected.Both NF-κB pathway and MAPK pathway were detected.Conclusions The in vitro FFA mediated steatotic cell model could induce the expression change of A20 and secretion of inflammatory cytokines.NF-κB and MAPK pathways involved in the response to FFA in HepG2 cells and U937 cells.

OBJECTIVE： To study the effects of Sinorhizobium fredii， Rhizobium radiobacter and Azorhizobium caulinodans on germination of Andrographis paniculata seed under drought and salt stress condition. METHODS： Drought and salt stress models of A. paniculata seed were established with PEG-6000 （called “PEG” for short）and NaCl respectively. The effects of different concentrations of PEG （0.05， 0.10， 0.15， 0.20， 0.25 g/mL） and NaCl （50， 100， 150， 200 mmol/L） on the germination indicators （germination rate， germination potential， germination index， vigor index） of A. paniculata seed were investigated by the method of germination on dish paper. The seeds were soaked with S. fredii， R. radiobacter and A. caulinodans， and then germination indicators were investigated under the condition of drought and salt stress. RESULTS： Drought and salt stress models of A. paniculata seed were established with 0.15 g/mL PEG solution and 50 mmol/L NaCl solution， respectively. S. fredii pretreatment could significantly improve all the germination indicators of A. paniculata seed under drought stress condition， as well as germination index and vigor index of it under salt stress condition， but significantly reduce germination rate and germination index of it under salt stress condition （P＜0.05）. R. radiobacter treatment could significantly improve all the germination indicators of A. paniculata seed under drought stress condition， as well as germination potential， germination index and vigor index of it under salt stress condition （P＜0.05）. A. caulinodans pretreatment could significantly improve germination rate， germination index and vigor index of A. paniculata seed under drought stress condition， as well as germination rate and germination potential of it under salt stress condition （P＜0.05）. CONCLUSIONS： Three kinds of Rhizobia can improve germination ability of A. paniculata seed under drought or salt stress condition to different degrees. This research can provide technical support for planting and cultivation of A. paniculata under drought condition， and using saline-alkali soil as A. paniculata plantation area.

Background: Chronic exposure to elevated levels of saturated fatty acids results in pancreatic β-cell senescence. However, targets and effective agents for preventing stearic acid-induced β-cell senescence are still lacking. Although melatonin administration can protect β-cells against lipotoxicity through anti-senescence processes, the precise underlying mechanisms still need to be explored. Therefore, we investigated the anti-senescence effect of melatonin on stearic acid-treated mouse β-cells and elucidated the possible role of microRNAs in this process. Methods: β-Cell senescence was identified by measuring the expression of senescence-related genes and senescence-associated β-galactosidase staining. Gain- and loss-of-function approaches were used to investigate the involvement of microRNAs in stearic acid-evoked β-cell senescence and dysfunction. Bioinformatics analyses and luciferase reporter activity assays were applied to predict the direct targets of microRNAs. Results: Long-term exposure to a high concentration of stearic acid-induced senescence and upregulated miR-146a-5p and miR- 8114 expression in both mouse islets and β-TC6 cell lines. Melatonin effectively suppressed this process and reduced the levels of these two miRNAs. A remarkable reversibility of stearic acid-induced β-cell senescence and dysfunction was observed after silencing miR-146a-5p and miR-8114. Moreover, V-maf musculoaponeurotic fibrosarcoma oncogene homolog A (Mafa) was verified as a direct target of miR-146a-5p and miR-8114. Melatonin also significantly ameliorated senescence and dysfunction in miR-146a-5pand miR-8114-transfected β-cells. Conclusion These data demonstrate that melatonin protects against stearic acid-induced β-cell senescence by inhibiting miR-146a- 5p and miR-8114 and upregulating Mafa expression. This not only provides novel targets for preventing stearic acid-induced β-cell dysfunction, but also points to melatonin as a promising drug to combat type 2 diabetes progression.

Background::Abnormal lipids are strong predictors of cardiovascular disease in type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM). However, the potential associations of insulin resistance (IR) and beta-cell function (BCF) with abnormal lipids in newly diagnosed T1DM or T2DM patients are not fully understood.Methods::A cross-sectional survey of 15,928 participants was conducted. Homeostasis model assessment and postprandial C-peptide levels were used to estimate IR and BCF. A restricted cubic spline (RCS) nested in binary logistic regression was used to examine the associations of IR and BCF with abnormal lipids.Results::High triglyceride (TG), low high-density lipoprotein cholesterol, and high low-density lipoprotein cholesterol (LDL-C) accounted for 49.7%, 47.8%, and 59.2% of the participants, respectively. In multivariable analysis, high IR was associated with an increased risk of high TGs ( P for trend <0.001) in T1DM and is associated with an elevated risk of high TG and low HDL-C (all P for trend <0.01) in T2DM. Low BCF was not associated with risks of dyslipidemia in patients with T1DM or T2DM after adjustment for potential confounders. Conclusion::High IR had different associations with the risk of dyslipidemia in newly diagnosed T1DM and T2DM patients, suggesting that early treatment that improves IR may benefit abnormal lipid metabolism.

OBJECTIVE： To study the effects of 60Co-γ radiation on the botanical traits and property of Andrographis paniculata， and to screen suitable irradiation dose. METHODS： The seeds of A. paniculata were irradiated by 60Co-γ rays with different irradiation doses（0，10，20，50，100，200，300 Gy）. The botanical traits indexes of A. paniculata as seed germination rate， root length， seedling rate， seedling height， leaf area， fresh weight， dry weight， stomata number of lower epidermis of leaf， and its property indexes as the contents of andrographolide， dehydrated andrographolide and chlorophyll， activity of T-SOD enzyme and CuZn-SOD enzyme， were determined after radiation. The coefficient of variation （CV） was calculated. The linear regression analysis was performed for seedling rate， and medial lethal dose was calculated. The correlation analysis was performed between the parameters of botanical trait and quality property with irradiation dose. Cluster analysis was conducted for M1 generation of A. paniculata in different irradiation dose groups by connection method combined with squared euclidean distance. RESULTS： Different irradiation doses showed different effects on botanical traits and property of A. paniculata. According to the average value of CV， the index of botanical traits was ranked as leaf area ＞ fresh weight ＞ dry weight ＞ plant height ＞ root length ＞ stomata number ＞ seedling rate ＞ germination rate； among different irradiation dose groups， the coefficient of variation was ranked as 50 Gy＞200 Gy＞100 Gy＞20 Gy＞10 Gy＞300 Gy＞0 Gy. According to the average value of CV， the index of property was ranked as dehydrated andrographolide content＞andrographolide content＞chlorophyll content＞CuZn-SOD enzyme activity＞T-SOD enzyme activity； among different irradiation dose groups， the coefficient of variation was ranked as 100 Gy＞50 Gy＞200 Gy＞20 Gy＞10 Gy＞300 Gy＞0 Gy. The medial lethal dose was 195.10 Gy. According to the botanical traits， M1 generation of A. paniculata of 7 dose groups could be divided into 4 types. According to the property， M1 generation of A. paniculata of 7 dose groups could be divided into 3 types. CONCLUSIONS： The suitable irradiation dose interval for irradiating A. paniculata is 50-200 Gy.

Objectives:The purpose of this study is to evaluate the impact of the cumulative workload of HUTT testing physicians on diagnostic outcomes. Methods:This study retrospectively and consecutively included the data of testing physicians and patients who underwent HUTT at Fuwai Hospital,Chinese Academy of Medical Sciences,from January 2016 to December 2022.Based on the cumulative workload of physicians during the period from the initiation of tilt tests at the hospital to the end of the study,the physicians were categorized into low (50-100 sases),moderate (100-350 sases),and high (1000-4000 sases) cumulative workload groups,the cumulatie workload of no physician is 351-999 sases.Additionally,physicians were grouped by sex,educational background,and professional title to analyze differences in diagnostic rates of tilt table test reports within and between these groups. Results:The study included 22 testing physicians and 6122 patients.There were statistically significant differences in the rates of positive,suspicious positive,and negative reports among the 22 physicians (P<0.001).The average suspicious positive report rate in the moderate cumulative workload group was significantly higher than in the low and high cumulative workload groups (3.21％ vs.1.09％ vs.1.62％,P=0.001).The suspicious positive report rate was higher in the female physician group compared to the male physician group (2.25％ vs.1.07％,P=0.017),in the undergraduate physician group compared to the postgraduate physician group (2.46％ vs.1.52％,P=0.013),and in the junior title group compared to the intermediate and senior title groups (3.40％ vs.1.75％ vs.2.53％,P=0.024).Multivariate logistic regression analysis showed that a moderate cumulative workload was an influencing factor for suspicious positive reports,regardless of whether negative or positive was used as the reference (all P<0.05). Conclusions:There are certain differences in the diagnostic report rates of HUTT among different individual physicians.Physicians with a moderate cumulative workload are more likely to issue suspicious positive HUTT diagnostic reports.

Objective To explore the efficacy and safety of pars plan vitrectomy(PPV)combined with internal limit-ing membrane(ILM)covering surgery in patients with large-diameter idiopathic macular holes(IMH)using optical coher-ence tomography angiography(OCTA)and microperimetry.Methods Twenty-three eyes of 23 patients with large-diame-ter IMH treated at the Eye Hospital Affiliated with Nanchang University from February 2023 to July 2023 were selected.The patients'best-corrected visual acuity(BCVA),hole closure rate,microcirculation[perfusion density at the superficial cap-illary plexus(PSCP)and vascular density at the retinal superficial capillary plexus(VSCP)]in the macular area(divided into central,inner and outer regions),macular sensitivity(MS),fixation stability,and foveal avascular zone(FAZ)area were observed before surgery and at 1 week,1 month,3 months,and 6 months after surgery.Changes in eye parameters before and after surgery were compared,and the correlation between parameters was analyzed.Results After a 6-month follow-up,all macular holes in the affected eyes were closed,with a closure rate of 100％.Compared to before surgery,the BCVA of the patients improved gradually at 1 week,1 month,3 months,and 6 months after surgery,and the differ-ences between any two time points were statistically significant(all P＜0.05).The FAZ area decreased at 1 week,1 month,3 months,and 6 months after surgery compared to before surgery,and the differences were statistically significant(all P＜0.05).The VSCP in the three regions showed a decrease at 1 week postoperatively and an increase at 6 months postoperatively compared to before surgery,and the differences were statistically significant(both P＜0.05).The PSCP in-creased at 3 months and 6 months postoperatively compared to before surgery,with statistically significant differences(both P＜0.05).The 12° MS increased at 1 month,3 months,and 6 months after surgery compared to before surgery,with statistically significant differences(all P＜0.05).The proportions of fixation points within 2° and 4° increased at 1 month,3 months,and 6 months postoperatively compared to before surgery,with statistically significant differences(all P＜0.05).The proportion of fixation points within 4° increased at 1 week postoperatively compared to before surgery,with a statistically significant difference(P＜0.05).Correlation analysis showed that preoperative MS and the proportion of fixa-tion points within 2° were negatively correlated with BCVA(logMAR)at 6 months postoperatively(r=-0.819,-0.790,both P＜0.05),while preoperative FAZ area and minimum hole diameter were positively correlated with BCVA(logMAR)at 6 months postoperatively(r=0.596,0.853,both P＜0.05).The preoperative VSCP in the inner and outer regions were negatively correlated with BCVA(logMAR)at 6 months postoperatively(r=-0.521,-0.570,both P＜0.05).Conclu-sion The results of OCTA and microperimetry show that after treatment with PPV and ILM covering surgery,the closure rate of large-diameter IMH is good,and all indicators have improved compared to before surgery.

Objective:To observe the effectiveness and safety of pars plana vitrectomy (PPV) combined with inner limiting membrane (ILM) removal and 41G microneedle subretinal injection of balanced salt solution (BSS) in the treatment of refractory macular hole.Methods:A prospective clinical study. From January to June 2023, 20 cases (20 eyes) of refractory macular hole patients diagnosed through examination at The Affiliated Eye Hospital of Nanchang University were included in the study. The basal diameter of the affected eye's basal diameter (BD) was >1 000 μm. Macular hole index (MHI) was <0.5. The affected eye received treatment with 23G PPV combined with ILM removal and 41G microneedle subretinal injection of BSS. Best corrected visual acuity (BCVA), microperimetry, and optical coherence tomography angiography (OCTA) were performed before and 1, 2, 3, and 6 months after surgery for the affected eye. BCVA examination was performed using standard logarithmic visual acuity chart, and convert it to logarithmic minimum resolution angle (logMAR) visual acuity for statistical purposes. MP-3 microperimetry was used for micro view examination, record the mean sensitivity (MS) of the retinal within a 12° range of the fovea. OCTA was used to measure the area of the avascular zone of the macula (FAZ), perimeter of the FAZ (PERIM), retinal vascular length density (VLD), and vascular perfusion density (VPD). The changes in BCVA, MS, FAZ area, PERIM, VLD, VPD before and after surgery were compared and analyzed. After the same time, the closure of macular hole and the occurrence of complications after surgery were observed. Single factor analysis of variance was used to compare the observation indicators at different times before and after surgery. The correlation between various observation indicators and preoperative minimum diameter (MD), BD, and hiatus height at 6 months after surgery were analyzed using Pearson correlation analysis.Results:Among the 20 cases with 20 eyes, there were 2 males with 2 eyes and 18 females with 18 eyes. Age was (61.45±8.56) years old. The logMAR BCVA, MS, FAZ area, PERIM, VLD, and VPD of the affected eye were 1.46±0.21, (16.20±5.81) dB、(0.40±0.17) mm 2, (2.89±0.99) mm, (6.23±3.59) mm -1, (0.17±0.10)%, respectively. Six months after surgery, out of 20 eyes, macular hole closure and incomplete closure were 18 (90.0%, 18/20) and 2 (10.0%, 2/20) eyes, respectively. The logMAR BCVA, MS, FAZ area, PERIM, VLD, and VPD were 0.80±0.20, (22.20±4.60) dB, (0.18±0.10) mm 2, (1.83±0.80) mm, (9.54±2.88) mm -1, (0.31±0.14)%. Compared with before surgery, the differences were statistically significant ( P<0.05). The correlation analysis results showed a positive correlation ( P<0.05) between preoperative BD and postoperative 6-month PERIM and VPD. There was a negative correlation between preoperative MD and postoperative VLD at 6 months ( P<0.05). There was a negative correlation between preoperative MHI and logMAR BCVA and VPD at 6 months after surgery ( P<0.05). No complications such as elevated or decreased intraocular pressure, damage to retinal pigment epithelium, retinal hemorrhage, endophthalmitis, or retinal detachment occurred after surgery in all affected eyes. Conclusion:Minimally invasive PPV combined with ILM removal and 41G microneedle subretinal injection of BSS can effectively improve the closure rate of refractory macular hole patients in the short term, improve vision, and have good safety.