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.

Objective:Effects of Qingsui Xiaoyan Decoction on osteoclast differentiation of RAW264.7 cells.Methods:Receptor activator of nuclear factor kappa B ligand (RANKL) was used to induce differentiation of RAW264.7 cells into osteoclasts. CCK-8 method was used to screen experimental drug concentrations and analyze the cytotoxicity of drug intervention on RAW264.7 cells. TRAP staining method was used to quantitatively detect the effects of Qingsui Xiaoyan Decoction on osteoclast differentiation ability. RT-PCR technology was used to detect the mRNA expressions of osteoclast differentiation genes TRAP, MMP-9, CK, CTR. Western blot was used to detect osteoclast-related protein expressions of NF-κBP65, IκB-αTRAP, and TRAP.Results:After 72 hours of intervention, 0.800, 4.000, 20.000, and 100.000 mg/ml of Qingsui Xiaoyan Decoction could inhibit cell proliferation ( P<0.05). Compared with the model group, the number of TRAP-positive cells decreased ( P<0.05) in the 5, 20, 80, and 160 μg/ml Qingsui Xiaoyan Decoction groups. Additionally, the mRNA levels of TRAP, MMP-9, CK, and CTR decreased ( P<0.05), the expression of NF-κB p65 and TRAP proteins decreased ( P<0.05), and the expression of IκB-α protein increased ( P<0.05). Conclusions:Qingsui Xiaoyan Decoction can inhibit the expressions of TRAP, MMP-9, CK, CTR genes and TRAP protein, reduce the degradation of I κ B - α protein by inhibiting the NF - κ B signaling pathway, inhibit the expression of NF - κ B p65 protein, and thus inhibit the differentiation of RAW264.7 into osteoclasts.

Objective To investigate and validate the expression profiles of Ppp3cb and Ppm1g through differential proteomic analysis of hippocampal tissue in NHE1 gene knockout mice with proteomic analysis.Method ① Six 2-week-old NHE1 knockout mice were selected as the model group,and 6 wild-type mice of the same age served as the control group,and their genotypes were detected by agar-gel electrophoresis.Open field test and forced swimming test were used to evaluate the behaviors of mice in the model group and control group,and epileptic seizure was graded according to Racine scoring.② Tandem mass spectrometry was employed to screen the differential proteins in the hippocampus tissues from the model group and the control group.Then the obtained differential proteins were annotated and enriched in the Gene Ontology(GO)database.Search tool for the retrieval of interesting genes(STRING)database was used to analyze protein-protein interaction(PPI)among different proteins.③ The transcriptional and translational levels of Ppp3cb and Ppm1g were detected by qPCR and Western blotting,respectively,and their expression levels in the tissues were observed with immunohistochemistry.Results ① NHE1 was not expressed in the model group.The mice of the model group had shorter total movement distance(P=0.007 3)and less crossing cells(P＜0.000 1)in open field test,and longer period of immobility in forced swimming test(P＜0.000 1)when compared with those from the control group.② When fold change ≥1.2 times and P＜0.05 were set as the significant threshold for differential expression,845 differentially expressed protein sites were detected in the hippocampus,among which 9 proteins(including Ppm1g)were up-regulated and 7 ones(including Ppp3cb)were down-regulated.Gene Ontology(GO)functional analysis showed that after NHE1 knockout,the most significant differences were observed in the concentration of molecular function(MF)related to protein serine/threonine phosphatase activity,concentration of cellular component(CC)related to the plasma membrane,and concentration of biological process(BP)related to negative regulation of biological processes and immune system processes.STRING analysis indicated that the differential proteins Ppp3cb and Slc9a1 directly acted,Ppm1g indirectly acted through Ppp3cb and Slc9a1,and Ppp3cb and Ppm1g interacted.③The transcriptional and translational levels of Ppp3cb were decreased,and its expression level was reduced in the tissues,while those of Ppm1g were increased,and its expression was elevated in the tissues(P＜0.05).Conclusion In the hippocampus of NHE1 gene knockout mice,the expression of differential protein Ppp3cb is down-regulated and that of Ppm1g is up-regulated,which provide a basis for further study on their involvement in the pathogenesis of epilepsy.

【Objective】 To compare the difference in the detection rate of microorganisms in cord blood between BACTEC FX and BacT/ALERT 3D automated blood culture systems, and to compare the influence of incubation time and different types of culture sample on the detection rate of microorganisms in cord blood. 【Methods】 Cord blood samples prepared from April to August 2020 in Sichuan Cord Blood Bank(n=4 358) were selected, and 20 mL of plasma was used as culture samples for microbial detection. In addition, cord blood samples prepared in the same months of 2021(n=4 057) were selected, and 19 mL of plasma plus 1 mL of final product was used as culture samples for microbial detection. The total sample size was 8 415, of which 4 849 samples(2 458 in plasma group and 2 391 in plasma plus final product group) were assigned to the BACTEC FX system, and 3 566 samples(1 900 in the plasma group and 1 666 in the plasma plus final product group) to the BacT/ALERT 3D system. All samples were cultured for 7 days, and culture data were recorded on day 5 and day 7. Positive results were confirmed by Gram staining. 【Results】 The positive rate detected by the BACTEC FX system was higher than that of the BacT/ALERT 3D system(4.08% vs 2.69%), with statistically significant difference(P<0.05). The positive rates of BACTEC FX and BacT / ALERT 3D on day 7 increased by 8% and 4.26% respectively compared with on day 5. When the culture sample types were concerned, the BACTEC FX showed statistically significant difference in positive rate between plasma group and plasma plus final product group(P<0.05), while no significant difference was found in positive rate(P>0.05) detected by the BacT/ALERT 3D system. With quality control strains, there were significant differences in TTP between these two systems for Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Clostridium sporogenes, and Bacillus subtilis(P<0.05), but no significant difference for Aspergillus brasiliensis and Candida albicans(P>0.05). 【Conclusion】 This study suggests that the selection of BACTEC FX blood culture system with incubation time of not less than 7 days and plasma plus final product as culture samples may improve the detection rate of microorganisms in cord blood.

Background: Dietary fiber is strongly recommended as the basic treatment for functional constipation according to global guidelines. However, a complete evaluation standard for the laxative functional food remains to be improved in China. Aims: To evaluate the efficacy and safety of the laxative function of a compound fructose-oligosaccharide fiber granule, so as to provide evidence-based medical basis for the evaluation of laxative functional food. Methods: In a randomized, double-blind, parallel and controlled trial with placebo as control, 120 subjects with functional constipation were enrolled in 2 clinical research centers in Beijing, and randomly divided into experimental group and control group (60 cases in each group). Subjects in experimental group were given a compound fructose-oligosaccharide fiber granule dissolved in 50 mL water orally, 1 bag (9 g) per day for 2 weeks; while those in control group were given a placebo granule with the same appearance, specification and dosage as the experimental group. The bowel movement frequency per week, defecation status and stool consistency were recorded before and after the test, and the safety tests were completed. Results: After 2 weeks of treatment, the bowel movement frequency in experimental group increased by (1.63±1.57) times per week, the stool consistency assessed by Bristol stool form scale and the difficulty in defecation were also improved as compared with the baseline (all P<0.05). Furthermore, improvements in experimental group were superior to those in control group (all P<0.05). No allergic and other adverse events were reported during the test, and there were no significant changes in blood, urine, stool routine and blood biochemical indices before and after the test. Conclusions: The compound fructose-oligosaccharide fiber granule tested in this study is proved to have laxative effect and is safety for functional constipation. The testing program is scientific and of feasibility, and may provide a methodology basis for human oral administration trials of laxative functional food.

Objectives The aim of the study was to detect the frequency of B regulatory cells (Breg)and the correlation between Breg and T regulatory cells (Treg) in pancreatic cancer patients,and to investigate its role in pathogenesis of pancreatic cancer.Methods 50 pancreatic cancer patients and 21 healthy controls were enrolled.CD19+CD24hiCD38hi Breg and CD4+CD25high Treg was also determined via flow cytometry.The correlation between Breg and Treg was analyzed by Spearman correlation test.Results Upregulation of CD19+CD24hiCD381hi Breg was associated with pancreatic cancer progression.Furthermore,this B cell subset was positively correlated with the frequency of CD4 + CD25high Treg cells.Conclusions Together,CD19 + CD24hiCD38hi Breg cells are significantly elevated in pancreatic cancer patients,indicating this B cell subset might play an vital role in clinical progression of pancreatic cancer.The significant positive correlation between Breg and Treg may suggest CD 1TCD24hiCD38hi Breg are affecting tumor progression through Treg cells.

Objective To investigate the clinical effect of insulin on gestational diabetes mellitus(GDM).Methods GDM patients were randomly divided into group A and group B,group B were treated with short term intensive insulin pump therapy,and group A were treated with insulin pump in short term combined with biosynthetic human insulin therapy.Results The body mass index and diabetes family history rates pre-pregnancy in group B were significantly higher than group A(P<0.05).Logistic analysis showed that family history of BMI pre-pregnancy and diabetes family history were independent risk factors for GDM drug treatment(P<0.05).Compared with the group B,the improvement of blood glucose levels and the incidence of complications in patients with group A were better than the former(P<0.05).Conclusion Rational application of insulin can improve the efficacy of clinical treatment of GDM, reduce the incidence of complications, with high safety.

[ ABSTRACT] AIM:To investigate the effect of Ginkgo biloba extract ( GBE) on diabetic retinopathy ( DR) and its possible mechanism in rats .METHODS:Goto-Kakizaki ( GK) rats were used as a DR model, and were treated with different doses of GBE.Normal Wistar rats were used as the control.Blood glucose and retina barrier injury were analyzed, respectively.Ganglion cell apoptosis was detected by TUNEL staining.Moreover, the protein expression of nuclear factor E2-related factor 2( Nrf2) , ERK, Bcl-2 and P53, and ERK phosphorylation were examined by Western blot.RESULTS:GBE reduced blood glucose in the DR rats, attenuated retina barrier injury, and decreased the apoptosis of ganglion cells. Furthermore, the expression of Nrf2 and Bcl-2, and phosphorylation of ERK were increased after GBE treatment, whereas P53 expression was decreased.CONCLUSION:GBE protects ganglion cells against apoptosis in DR rats, which may be through activation of Nrf2/ERK pathway and regulating Bcl-2 and P53 expression.

OBJECTIVETo elucidate the epidemic condition and molecular subtyping of ciprofloxacin and cefotaxime co-resistant Salmonella Indiana (S. Indiana) isolated from retail chicken carcasses in six provinces of China.

METHODSA total of 2 647 Salmonella strains isolated from retail chicken carcasses collected from six provinces of China were subjected to antimicrobial susceptibility testing. All Salmonella isolates co-resistant to ciprofloxacin and cefotaxime were further characterized by serotyping, extended-spectrum beta-lactamases (ESBLs) producing strains screening and pulsed field gel electrophoresis (PFGE) typing.

RESULTSAmong 2 629 Salmonella isolates tested, 227 (8.52%) isolates were co-resistant to ciprofloxacin and ceftazidime/cefotaxime (Beijing: 11.67% (99/874), Jilin: 8.20% (60/726), Guangdong: 1.39% (7/502), Jiangsu: 15.61% (42/260), Shaanxi: 8.56% (16/186), Inner Mongolia: 0 (0/81)), and 224 of them were identified as S. Indiana. 213 (95.10%) isolates of S. Indiana were ESBLs producing strains. All ciprofloxacin and cefotaxime co-resistant S. Indiana isolates developed a multi-drug resistant profile and 17.86% (40/224) of them were resistant to all antibiotics tested except carbapenems, and 50.89% (114/224) of them resistant to 9 antibiotics, additionally, 25.45% (57/224) of them showed multi-drug resistance to 8 antibiotics. All ciprofloxacin and cefotaxime co-resistant S. Indiana isolates were divided into 32 PFGE clusters and 150 PFGE patterns. Strains of S. Indiana from same or different sampling site and time seemed to either share the same PFGE patterns or be differential to each other in different regions.

CONCLUSIONThe results indicated that chicken carcasses collected from parts of China were heavily contaminated by ciprofloxacin and cefotaxime co-resistant S. Indiana and could serve as an important reservoir of ciprofloxacin and cefotaxime co-resistant Salmonella. Molecular subtyping results indicated that cross contamination or common pollution source might be in these strains.

Animals ; Anti-Bacterial Agents ; pharmacology ; Cefotaxime ; pharmacology ; Chickens ; microbiology ; China ; Ciprofloxacin ; pharmacology ; Drug Resistance, Multiple, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Food Contamination ; Food Microbiology ; Meat ; microbiology ; Salmonella ; classification ; isolation & purification ; Serotyping ; beta-Lactamases

Objective To observe the clinical effects of self-prescribed Anshen pills in the treatment of female patients with menopausal syndrome . Methods From August 2012 to September 2014, 100 female patients with menopausal syndrome in Tianjin Beichen District Chinese Medicine were randomly divided into control group (50 cases) and observation group (50 cases).The control group received tibolone orally, and the observation group received self-prescribed Anshen pills orally.All treated for 3 months.Modified Kupperman score, self-rating anxiety scale ( SAS) score, self-rating depression scale (SDS) score, Pittsburgh sleep quality index (PSQI) score, menopause-specific quality of life (MENQOL) score, the main clinical symptoms and hormone levels in serum were compared; the adverse reactions were recorded.Results Compared with control group post-treatment, hot flashes and perspiration, insominia improved greatly in observation group.The serum levels of luteinizing hormone ( LH ) in observation group was (28.04 ±3.37)IU/L, which was higher than (24.48 ±6.63)IU/L in control group.PSQI score in observation group was (6.57 ±1.23), which was lower than (8.18 ±1.70) in control group.The incidence of adverse reactions in observation group was 4.0%, which was lower than 18.0% in control group, and there were significant differences between two groups ( P<0.05 ) .Conclusion Self-prescribed Anshen pills is effective and safe for menopausal syndrome.