AIM:To observe the effect of levcromakalim(Lev) on endthelin-1(ET-1) induced proliferation and the expression of protein kinase C(PKC) in cultured rat vascular smooth muscle cells(VSMCs).METHODS: VSMCs were isolated from rat aorta and were cultured with different concentrations of Lev in vitro after stimulated to proliferation by ET-1(10~(-8)(mol?L~(-1))).Vitality of VSMCs was detected by MTT;DNA replication was evaluated by 3H-TdR method.Flow cytometry was used to analyze cell cycle and apoptosis.The expression of PKC? protein and mRNA were determined by Western blot and reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Vitality of VSMCs and 3H-TdR intermingle rate were decreased(P

Objective To investigate characteristics of free fatty acid (FFA) and high-sensitivity Creactive protein (hs-CRP) in patients with type 2 diabetes mellitus and metabolic syndrome (MS) compared to those with type 2 diabetes mellitus without MS.Methods Totally 120 patients with type 2 diabetes mellitus who received blood glucose control in Shaoxing People's Hospital from January to June 2013 were recruited and divided into MS group (n =56) and non-MS group (n =64).The serum FFA profile of the patients was measured by enzymatic assay, and the serum hs-CRP level measured by particle-enhanced immuno-precipitation assay.Results The level of FFA was higher in the MS group than that in the non-MS group [(0.60 ±0.25) mmoL/L vs.(0.45 ±0.21) mmol/L, P =0.033].The level of hs-CRP was higher in the MS group than that in the non-MS group [5.29 (4.69-5.82) mg/L vs.0.73 (0.42-1.26) mg/L, P =0.000].The level of hs-CRP was higher in the patients with central obesity than that in those without central obesity [4.34 (0.91-5.46) mg/L vs.1.80 (0.82-3.27) mg/L, P=0.014], also higher in hypertensive patients than in non-hypertensive patients [5.21 (3.18-5.96) mg/L vs.2.93 (0.89-4.98) mg/L, P =0.012].Conclusions Serum FFA and hs-CRP concentrations may be significantly higher in type 2 diabetes mellitus patients with MS.The increased levels of FFA and hs-CRP may play an important role in the pathogenesis of type 2 diabetes mellitus and MS.

Objective To investigate the relationship between triglyceride and serum polyunsaturated fatty acid (PUFA) in patients with metabolic syndrome (MS).Methods Totally 74 MS patients (MS group) and 62 healthy subjects (normal control group,NC group) were recruited from June 2011 to June 2012 in Shaoxing People's Hospital.Serum phospholipid fatty acid profiles were analyzed with capillary gas chromatography.Serum lipids were measured by enzymatic assay.Results The triglyceride concentration was significantly higher in MS group than that in NC group [(2.0 ±0.8) mmol/L vs.(1.3 ±0.5) mmol/L,P=0.000].The levels of n-3 PUFA,n-6 PUFA,and PUFA were significantly lower in MS group than those in NC group (9.8％ ±2.2％ vs.11.1％ ±2.4％,P=0.002; 35.4％ ±6.7％ vs.39.5％ ±7.8％,P=0.009; 45.2％ ±8.9％ vs.50.6％ ±10.1％,P =0.000).Triglyceride concentration was inversely correlated with n-3 PUFA and PUFA levels (r =-0.42,P =0.008 ; r =-0.23,P =0.013).Conclusions n-3 PUFA,n-6 PUFA,and PUFA decrease in MS patients.Triglyceride concentration is inversely correlated with PUFA.

Objective To investigate the relationship between serum phospholipid polyunsaturated fatty acids (PUFA) and interleukin-6 (IL-6) in patients with metabolic syndrome.Methods Serum phospholipid PUFA profiles of 61 patients with metabolic syndrome and 55 healthy controls was analyzed with high performance gas chromatography.Enzyme-linked immunosorbent assay was used to measure serum IL-6 levels.Results The IL-6 concentration was significantly higher in the MS group than in the control group [(16.0 ± 4.7) pg/ml vs.(9.2 ± 2.6) pg/ml,P =0.000].The total PUFA percentage was lower in the MS group than in the control group [(47.2±2.4)％ vs.(50.8 ±2.5)％,P=0.001].The serum IL-6 concentration was negatively correlated with n-3 PUFA,n-6 PUFA,and total PUFA (r =-0.51,P ＜0.01;r=-0.27,P＜0.01;r=-0.38,P＜0.01).Conclusions Serum PUFA in patients with metabolic syndrome is lower than in healthy people.Serum IL-6 concentration may be negatively correlated with total PUFA concentration.

Objective: To explore the clinicopathological features and the renal biopsy process of a case of IgG4-related chronic interstitial nephritis with perirenal capsule involved and review associated literature to improve the clinician's understanding for this disease and to perform a better renal biopsy. Methods: The onset, diagnosis and treatment course of the disease were described and associated literature were reviewed to summary the clinicopathologic features and key points in renal biopsy. Results: The data of the patient showed that the urine specific gravity was 1.011, with urine protein ± and urine sugar 3+. The concentration of hemoglobin was 53 g/L, serum creatinine was 1665 μmol/L, and IgG4 was 9.39 g/L. Computed tomography showed that both kidneys enlarged slightly with decreased density and low density shadow around the kidneys. On contrast-enhanced scan, irregular low-density enhancement areas were found in both kidneys, and the edge of the boundary was not clear. For the first renal biopsy, no renal parenchyma was found except mainly hyaline collagen fibrils. At the second time, 3 pieces of tissues were obtained, which showed chronic interstitial glomerulonephritis. The IgG4 positive plasma cells were about 60/HPF and the IgG4⁺/IgG⁺cells ratio was more than 40%. The diagnosis of IgG4-related chronic interstitial glomerulonephritis was confirmed. After corticosteroid treatment, the serum creatinine decreased to 502 μmol/L after the patient got rid of dialysis. Conclusions There are various manifestations of renal damage caused by IgG4-related disease. It is necessary to pay attention to the involvement of the perirenal capsule, and to balance the risk of bleeding and poor sampling in renal biopsy.

Objective:To explore the relationship between carotid atherosclerosis (CAS) and bone metabolism marker osteocalcin (OC) in patients with type 2 diabetes (T2DM).Methods:A total of 100 patients with T2DM admitted to Shaoxing People's Hospital (Shaoxing Hospital, Zhejiang University) from January 2018 to August 2018 were selected as study subjects, and the carotid intima-media thickness (IMT) was detected.The patients were divided into CAS group and normal carotid IMT group (NC group), with 50 cases in each group.The levels of OC, fasting plasma glucose(FPG), fasting insulin(FINS), glycosylated hemoglobin(HbA1c), total cholesterol (TC), triglyceride (TG), high density lipoprotein(HDL-C), low density lipoprotein (LDL-C), insulin resistance index (HOMA-IR) were compared between the two groups.Results:The OC level of the CAS group[(11.86±4.46)ng/mL] was significantly lower than that of the NC group[(23.94±4.52)ng/mL] ( t=-9.640, P=0.001). The LDL-C level of the CAS group[(2.89±0.82)mmol/L] was significantly higher than that of the NC group[(2.55±1.16)mmol/L]( t=2.03, P=0.049). Pearson correlation analysis showed that IMT was positively correlated with age, LDL-C, HbA1c ( r=0.285, 0.190, 0.173; P=0.000, 0.020, 0.035), and negatively correlated with OC ( r=-0.603, P=0.000). Conclusion:CAS in patients with T2DM is closely related to OC, and the reduction of OC levels may be a risk factor for CAS in T2DM patients.

Objective:To assess the bacterial profiles and antimicrobial susceptibility patterns in uropathogens, and help to inform the empiric treatment decisions for urinary tract infection in outpatient settings.Methods:A single institutional retrospective analysis was performed on positive urine cultures from outpatient settings between January 1998 and December 2018. To analyze changes over time, trends analysis were undertaken on bacterial profiles, antimicrobial susceptibility and resistance.Results:A total of 1.172 pathogenic bacteria were isolated after exclusion of duplicate strains originated from the same patient, including 991(84.6%) Gram-negative bacterial strains and 181(15.4%) Gram-positive strains. The most common Gram-negative uropathogens were Escherichia coli (60.8%) and Klebsiella pneumonia (8.1%). Enterococcus faecalis (4.6%) was the predominant Gram-positive strain. The detection rate of Escherichia coli increased significantly, from 50.8% to 63.2% ( χ2=7.978, P=0.046), and no significant difference was observed in the distribution of major uropathogenic bacteria over the 20 years (all P>0.05). The proportion of extended-spectrum β-lactamase (ESBLs) producing strains increased significantly across the 20 years ( P<0.05). The resistance rates of Escherichia coli to amoxicillin and clavulanate potassium, aztreonam, ceftazidime, ciprofloxacin and sulbactam + cefoperazone increased significantly (all P<0.05). All the isolates sustained high susceptibility to tazobactam + piperacillin, amikacin, imipenem and nitrofurantoin (95.0%, 95.7%, 97.9% and 91.1%). Similar to those of Escherichia coli, Klebsiella pneumoniae remained a high and stable sensitivity to tazobactam+piperacillin, amikacin and imipenem during the 20 years (79.1%, 88.0% and 80.3%). However, the proportion of ESBLs producing strains increased significantly ( P<0.05). Among Gram-positive bacteria isolates, the sensitivity rates of Enterococcus faecalis to ampicillin, nitrofurantoin and penicillin G were 100.0%. No vancomycin resistant strain was detected in Gram-positive bacteria. Conclusions:From 1998 to 2018, Escherichia coli and Klebsiella pneumoniae are the most common Gram-negative bacteria uropathogens obtained in outpatient settings. Significant increases of resistance to some antimicrobial agents such as second- and third-generation cephalosporins and fluoroquinolones are observed during the 20 years and high susceptibilities to tazobactam+piperacillin, amikacin, imipenem and nitrofurantoin sustain over time. Local treatment strategies of urinary tract infections on outpatient basis should be made according to epidemiology of drug resistance and individual characteristics to control the spread and curb the prevalence of drug resistant.

Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.