[Analysis of gene characteristics and core genome characteristics of carbapenem-resistant Enterobacter cloacae in rural residents of Weifang City, Shandong Province].

Objective: To investigate the drug-resistant gene characteristics and core genome characteristics of carbapenem-resistant Enterobacter cloacae (CR-ECL) in rural residents of Weifang City, Shandong Province. Methods: Fecal samples were collected from rural community residents in Weifang City, Shandong Province in 2017. Drug-resistant strains were screened using a carbapenem-resistant enterobacter chromogenic medium. CR-ECL positive strains were acquired via Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry(MALDI-TOFMS) analysis. The antibiotic resistance phenotype of CR-ECL was determined using a microbroth dilution assay. Whole genome sequencing (WGS) and analysis were conducted, along with an examination of the immediate vicinity of the blaNDM gene and phylogenetic analysis of the strains. Results: A total of 628 fecal samples were collected and tested, of which 6 were CR-ECL positive (detection rate 0.96%), all exhibiting multiple drug resistance (MDR) phenotypes. Six CR-ECL strains had four MLST genotypes (ST), all of which carried multiple drug resistance genes (blaNDM-1, blaNDM-5, etc.) and virulence genes (acrA, acrB, entB, fepC, etc.). There were mobile genetic elements ISAba125, TN3-IS3000, TN3 and IS5 in the genetic environment surrounding the blaNDM gene. The phylogenetic tree showed that the multi-locus sequence typing of the core genome (cgMLST) was consistent with the single nucleotide polymorphism (SNPs) results. The cgMLST results showed that the allele differences between strains 2BC0101B and 2BC0251B, 2BG0561B and 2BI0221B were 2 and 1, respectively. The SNPs results showed that the above two pairs of bacteria also clustered together. It was found that the strains of chicken fecal samples in the National Center for Biotechnology Information (NCBI) database were located in the center of the evolutionary tree, and the local sequences could be traced back to American human sequences. Conclusion: Multidrug-resistant CR-ECL is detected in rural community residents in Weifang City, Shandong Province.

Effects of PPARγ agonist pioglitazone on cardiac fibrosis in diabetic mice by regulating PTEN/AKT/FAK pathway.

OBJECTIVE: The aim of this study was to explore the role of pioglitazone (PIO), a peroxisome proliferator-activated receptor-gamma (PPARγ) agonist, in cardiac fibrosis of diabetic mice. MATERIALS AND METHODS: A total of 60 adult male C57/B6 mice were divided into 3 groups using a random number table, namely, control group (Sham group, n=20), diabetic cardiomyopathy group (DCM group, n=20), DCM + PIO group (n=20). Streptozocin (STZ) was injected into mice at a dose of 125 mg/Kg to induce the model of diabetes in vivo. After successful induction, mice in DCM + PIO group were intragastrically given PIO at 10 mg/kg/d once a day for 6 weeks. Meanwhile, those in Sham group and DCM group were given the same volume of normal saline. After 6 weeks, ejection fraction % (EF%), fraction shortening % (FS%) and heart rate of mice in each group were examined via echocardiography. Picrosirius red (PSR) staining assay was conducted to detect collagen deposition in myocardial tissues of mice in each group. The protein expression level of PPARγ in mouse myocardial tissues in each group was measured through Western blotting and immunohistochemical staining assays. Hematoxylin-eosin (H&E) staining assay was carried out to evaluate the myocardial hypertrophy of mice in each group. The protein expression level of transforming growth factor-ß (TGF-ß) in mouse myocardial tissues in each group was measured through immunohistochemical staining assay. In addition, Western blotting was employed to detect the expression of proteins related to the phosphate and tension homology deleted on chromsome ten (PTEN)/protein kinase B (AKT)/focal adhesion kinase (FAK) signaling pathway in myocardial tissues of mice in each group. RESULTS: The messenger ribonucleic acid (mRNA) and protein expression levels of PPARγ in mouse myocardial tissues were significantly lower in DCM group than those in Sham group (p<0.05). PPARγ agonist PIO could significantly increase the protein expression of PPARγ in myocardial tissues of DCM mice. The results of cardiac Doppler ultrasound revealed that PIO significantly upregulated EF% and FS% in DCM mice (p<0.05). Besides, PIO remarkably reduced collagen deposition and TGF-ß protein expression in myocardial tissues in DCM mice (p<0.05). H&E staining results showed that PIO notably attenuated myocardial hypertrophy in DCM mice (p<0.05). Furthermore, it was discovered that PIO markedly elevated PTEN protein in myocardial tissues of DCM mice and inhibited the phosphorylation of AKT and FAK proteins (p<0.05). CONCLUSIONS: The protective effect of PIO against cardiac fibrosis in diabetic mice may be related to its regulation on the PTEN/AKT/FAK signaling pathway. Our findings suggest that PIO is expected to become a targeted drug for the treatment of DCM in clinical practice.

Study of expression levels and clinical significance of miR-503 and miR-375 in patients with esophageal squamous cell carcinoma.

OBJECTIVE: This study aims to investigate the expression levels and clinical significance of miR-503 and miR-375 in patients with esophageal squamous cell carcinoma. PATIENTS AND METHODS: 40 cases of cancer tissues and adjacent tissues in patients with esophageal squamous cell carcinoma were collected from March 2017 to September 2017 in the Department of Gastroenterology in the First Affiliated Hospital of Henan Polytechnic University. qRT-PCR was used to analyze the expressions of miR-503 and miR-375 in cancer tissues and adjacent normal tissues. The association between their expressions and the gender, age, degree of tumor differentiation, TNM staging, presence or absence of lymph node metastasis, and other clinicopathological characteristics of patients with esophageal squamous cell carcinoma was analyzed. RESULTS: The results showed that the expression level of miR-503 in esophageal squamous cell carcinoma tissues (6.83±2.14) was significantly higher than that in adjacent tissues (2.45±1.13). The expression level of miR-375 in esophageal squamous cell carcinoma tissues (3.75±1.06) was significantly lower than that in adjacent tissues (7.45±1.13). The expression levels of miR-503 and miR-375 in esophageal squamous cell carcinoma were related to the existence of lymph node metastasis, degree of differentiation of esophageal squamous cell carcinoma and TNM staging (p<0.001). There was no correlation between the expressions of miR-503 and miR-375 and the age, gender and tumor size of patients (p>0.05). CONCLUSIONS: miR-503 was highly expressed in esophageal squamous cell carcinoma and miR-375 was lowly expressed in esophageal squamous cell carcinoma. miR-503 and miR-375 were closely related to the lymphatic metastasis, degree of differentiation and TNM staging of the tumor.

Effect of Selenium Against Lead-Induced Damage on the Gene Expression of Heat Shock Proteins and Inflammatory Cytokines in Peripheral Blood Lymphocytes of Chickens.

The possible beneficial role of selenium (Se) in heat shock proteins (HSPs) and inflammation damage induced by lead (Pb) in chickens is unclear. Therefore, the aim of this study was to investigate the effect of Se against Pb on the messenger RNA (mRNA) expression levels of HSPs (HSP 27, 40, 60, 70, and 90); heme oxygenase-1 (HO-1); and the inflammatory cytokines nuclear factor kappa B (NF-κB), tumor necrosis factor alpha (TNF-α), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) in the peripheral blood lymphocytes of chickens. A total of 360 1-day-old broiler chickens were randomly allocated into four groups (n = 90/group). The control group was fed a basic diet containing 0.2 mg/kg Se and 0.5 mg/kg Pb; the Se supplementation group (+Se group) was fed a Se-adequate (sodium selenite) diet containing 1 mg/kg Se and 0.5 mg/kg Pb; the Pb-supplemented group (+Pb group) was fed a Pb acetate diet containing 0.2 mg/kg Se and 350 mg/kg Pb; and the Se and Pb compound group (Se + Pb group) was fed a diet containing 1 mg/kg Se and 350 mg/kg Pb. The blood was collected and examined for the mRNA levels of HSP and inflammatory cytokine genes at 30 and 60 days old. The results showed that Pb poisoning induced the mRNA expression of HSPs and inflammatory cytokines in the peripheral blood lymphocytes of chickens. In addition, Se alleviated the Pb-induced increase in HSP and inflammatory cytokine mRNA levels in chicken peripheral blood lymphocytes. In conclusion, Se can antagonize the toxic effects of Pb on chickens and protect the chickens' peripheral blood lymphocytes in normal physiological function.

High percentage inorganic arsenic content of mining impacted and nonimpacted Chinese rice.

Two approaches were undertaken to characterize the arsenic (As) content of Chinese rice. First, a national market basket survey (n = 240) was conducted in provincial capitals, sourcing grain from China's premier rice production areas. Second, to reflect rural diets, paddy rice (n = 195) directly from farmers fields were collected from three regions in Hunan, a key rice producing province located in southern China. Two of the sites were within mining and smeltery districts, and the third was devoid of large-scale metal processing industries. Arsenic levels were determined in all the samples while a subset (n = 33) were characterized for As species, using a new simple and rapid extraction method suitable for use with Hamilton PRP-X100 anion exchange columns and HPLC-ICP-MS. The vast majority (85%) of the market rice grains possessed total As levels < 150 ng g(-1). The rice collected from mine-impacted regions, however, were found to be highly enriched in As, reaching concentrations of up to 624 ng g(-1). Inorganic As (As(i)) was the predominant species detected in all of the speciated grain, with As(i) levels in some samples exceeding 300 ng g(-1). The As(i) concentration in polished and unpolished Chinese rice was successfully predicted from total As levels. The mean baseline concentrations for As(i) in Chinese market rice based on this survey were estimated to be 96 ng g(-1) while levels in mine-impacted areas were higher with ca. 50% of the rice in one region predicted to fail the national standard.

[Determination of salicylic acid, resorcinol and chloramphenicol in junlieping tincture by reversed-phase high performance liquid chromatography].

A reversed-phase high performance liquid chromatographic method was developed for the quantitative determination of salicylic acid, resorcinol and chloramphenicol in Junlieping tincture. The operation was carried out on an ODS column with the mobile phase of a mixture of acetonitrile-methanol-water(25:10:65, volume proportion). The eluate was detected at 278 nm wavelength. This method is simple, fast and can be used for the quality control of Junlieping tincture.

[Study of environmental pollution and damage of cytogenetic materials in urban residents].

Recently, epidemiologic data have indicated that the number of deaths caused by malignant tumor presented a significantly rise in Beicun countryside of Datong city. In order to understand the causes of high incidence of cancer, factors were investigated and analysed related to air, river water, drinking water, vegetables and human serum. Results showed that Benzo (a)-pyrene (Bap), total suspended particulates (TSP) in air, nitrate, nitrite in drinking water and vegetables were all significantly higher in Beicun countryside region than those in controlled countryside region, respectively (P < 0.01-P < 0.001). The micronuclei (MN) and berration nucleus (AN) rates of cultured lymphocytes were significantly higher while cpm value of unscheduled DNA synthesis (UDS) test were significantly lower than those in controlled region among residents in Beicun countryside region, respectively (P < 0.01-P < 0.001). The micronuclei, abrerration nucleus rates and cpm value of cultured lymphocytes among residents in Beicun countryside were strongly correlated with Bap, TSP, nitrite and nitrate contents in determination samples. Our results suggest that the air and water source under serious pollution may serve as an important factor causing cytogenetic damage among residents and cancer death rise in Beicun countryside.

[Curative effect of combined traditional Chinese medicine with Western medicine therapy on chronic nephritis and its correlation with hemorheology and microcirculation].

Patients were divided into two groups, 89 cases treated by combined traditional Chinese and Western medicine (TCM-WM) therapy, the other 36 served as control. Each of two groups was further subdivided into 4 clinical types and 5 Syndromes of TCM. Hemorheology and microcirculation were tested before and after treatment, the clinical curative effects of both groups were compared and correlation analysis between clinical curative effect and improvement of hemorheology and microcirculation was made. The results showed that the remission rate and total effective rate were 79.8% and 92.1% respectively in testing group, significantly higher than those (25% and 50%) of control (P < 0.05-0.01). The curative effect on each clinical type and Syndrome of TCM was better than that on corresponding type and Syndrome of control (P < 0.05-0.01). The improvement of hemorheology and microcirculation was more significant in testing group than that in control. It was very significant in complete and partial remission of both group (P < 0.05-0.01) but not marked in the cases of improved and ineffective cases (P > 0.05). Therefore clinical curative effect was closely related with them. So any treatment which could improve hemorheology and microcirculation would be useful in treating chronic nephritis.

Where does transformation occur in acute leukemia?

We studied 43 consecutive cases of acute leukemia for evidence of hybrid leukemia including biphenotypic or bilineal involvement. Twenty-two were initially diagnosed as acute lymphoblastic leukemia (ALL) and 21 as acute myelogenous leukemia (AML). Techniques included morphology, cytochemistry, immune phenotyping and cytogenetics. Thirty-one cases seemed restricted to one lineage. Twelve cases showed involvement of both lymphoid and myeloid cells. Dual staining immune phenotyping showed coexpression of diverse lineage markers. These data indicate a considerable proportion of unselected cases of acute leukemia are hybrid leukemias. These data are consistent with the notion that transformation frequently occurs in a stem or progenitor cell.

Hepatic function and drug pharmacokinetics after total body irradiation plus bone marrow transplant.

Radiation nephritis is the principle late toxicity seen after total body irradiation in barrier-maintained rats when hematologic toxicity is prevented by bone marrow transplantation. Renal dysfunction is observed for single doses as low as 7.5 Gy. Hepatic blood flow, as measured by indocyanine green clearance, is decreased after 8.5-9.5 Gy single-dose total body irradiation. Serum albumin levels are decreased after 9.5 Gy single-dose total body irradiation. Hypoalbuminemia is a symptom of hepatic damage, but can also be caused by renal damage or edema. No decrease in total serum protein is observed, indicating that proteinuria resulting from renal damage is not the cause of hypoalbuminemia. No edema and some dehydration are observed. These data indicate that hepatic damage as well as renal damage may be occurring after total body irradiation plus bone marrow transplantation. Animals given total body irradiation plus bone marrow transplantation show decreased tolerance to a wide variety of immunosuppressive and cytotoxic drugs, even when these drugs are given months after total body irradiation. Altered drug clearance after total body irradiation plus bone marrow transplantation is observed for cis-platinum, vincristine, and adriamycin. The increase in cis-platinum toxicity after total body irradiation plus bone marrow transplantation is caused by decreased renal drug clearance. The decrease in vincristine tolerance and the alterations in adriamycin and vincristine pharmacokinetics are caused by altered drug distribution after total body irradiation plus bone marrow transplantation. These results indicate that bone marrow transplant survivors may show altered clearance of, and decreased tolerance to, a wide variety of drugs that are used after bone marrow transplantation.

Are some cases of acute leukemia with t(8;21) hybrid leukemias?

Translocations between chromosomes 8 and 21, t(8;21)(q22;q22), occur most commonly in acute myelogenous leukemia (AML) of the M2 FAB type. We studied two cases of acute leukemia with t(8;21) by immune phenotyping and IgH and T-cell receptor beta chain gene rearrangement analyses. These cases had increased blasts in bone marrow (greater than 50%). Auer rods, and evidence of granulocyte maturation. Blasts from both cases expressed CD19(B4), a B-cell antigen, as well as myeloid antigens including CD13(My7) and CD33(My9). HLA-DR, CD34, and TdT were also strongly positive. IgH or TCR beta gene rearrangements were not detected. We suggest that some cases of acute leukemia with t(8;21) may be hybrid leukemias with transformation in a multipotent stem cell.

Use of conditioned media in cell culture can mask cytogenetic abnormalities in acute leukemia.

Conditioned media (CM) from a human lung adenocarcinoma cell line expressing interleukins 1 and 6 (IL-1, IL-6), granulocyte (G), macrophage (M), and GM colony-stimulating factors (G, M, GM-CSF) and transforming growth factor beta (TGF beta) were used to stimulate growth of bone marrow (BM) cells from 18 persons with leukemia, myelodysplastic syndrome, or lymphoma. The objective was to increase numbers of analyzable metaphases and to enhance the likelihood of detecting cytogenetic abnormalities. Although more mitotic cells were observed with CM, the detection rate of cytogenetic abnormalities decreased in 12 of 18 cases. These data indicate that use of CM for cytogenetic analyses may favor growth of normal versus leukemia cells and mask cytogenetic abnormalities.

[Quantitative changes in mitochondrial membrane structure of leukemic cells in children with acute leukemia].

The quantitative changes of mitochondrial membrane of AL cells were studied by morphometric technique under electron microscope. We found that the delta sM was 25.0% more increased and the delta mM was 18.0% more decreased in AL than in the controls. The extent of changed of delta mM in C-ALL and N-ALL was much greater than that in T-ALL. We also found that the changing extent of delta mM in AL is related with long term disease-free survival. During the period of inductive therapy, observation on the dynamic changes of delta mM may provide a useful information for predicting the therapeutic effects and the therapeutic scheme individually.

Characteristic of marrow culture and clinical relevance in leukemia with 8/21 translocation.

Karyotypes and in vitro cultures of bone marrow cells from eleven patients with acute myelogenous leukemia M2 type were performed. 8/21 translocation was found in 5 patients. In all of them the cultures in vitro shared one characteristic, i.e., clusters were small and less in number. These patients could relatively easily obtain complete remission. The similarity of CFU-GM in patients with t (8; 21) may be another important biological phenotype.