Objective To investigate the concentration change of indoxyl sulfate (IS) in blood and the renal expression of renal fibrosis-related factors (transforming growth factor-beta 1, TGF-β1;fibronectin, FN) after administration of mutagenic lactobacilli by oral.Methods A total of 60 male Sprague-Dawley (SD) rats aged 6 weeks was divided randomly into 3 groups.The normal control group (Sham group, n =20) received Sham operation of just incision of skin without kidney removed.The other two groups of rats were renal failure models selected from survivals of the other 40 rats who received real operation with 5/6 of kidney removed.Finally, 35 survived renal-failure rats were divided randomly into 2 groups : pathological control group(Model group, n =17) who were administrated of 2ml sterile saline solution once a day by gavage, and experimental group (lactobacillus bulgaricus (LB) group, n =18) who were administrated of 2 ml mutagenic lactobacilli (1.5 × 108 cfu/ml) once a day by gavage.Eight weeks later, blood specimens were taken to test the concentration of IS with high performance liquid chromatography-fluorescence detection (HPLC-FLU), and urea and creatinine by automatic biochemical analyzer;moreover, the rats were killed to get kidney tissues for pathological examination.Results The levels of serum IS, urea, and creatinine were statistically significantly different between two groups (P ＜ 0.05).Both the levels of renal tubular damage and renal interstitial fibrosis were both lessen in the experimental group compared to the model group (P ＜0.05).TGF-β1 and FN expressions in renal tissues were significantly decreased (P ＜0.05).Conclusions Mutagenic lactobacilli not only reduces serum concentration of IS, urea and creatinine in renal failure rats but lowers the expressions of TGF-β1 and FN in renal tissues.

Objective In this study, the glucose and energy metabolism-related genes (PGC-1α, Glut-4, ERRα, NRF-1, TFAM and mtDNA gene) were detected in type 2 diabetes mellitus ( T2DM) and non-T2DM minipigs, and the gene function was explored for T2DM pathogenesis.Methods The longissimus muscle of T2DM and non-T2DM Guangxi Bama mini-pigs was used as experiment material.The expression of glucose and energy metabolism-related genes was detec-ted by QRT-PCR.Results The expressions of PGC-1α, Glut-4, ERRαand NRF-1 genes were significantly higher than that of non-T2DM group, the expressions of TFAM and mtDNA gene were lower than that of non-T2DM group.Conclu-sions The upregulated expression of PGC-1αgene and its downstream genes Glut-4, ERRα, NRF-1 may improve the glu-cose metabolic functions in skeletal muscle in the Bama minipigs, whereas insufficient mitochondrial synthesis may induce decreasing ATP synthesis, and results in skeletal muscle insulin resistance, finally leading to the T2DM occurrence.

Objective To clone the coding sequence of Guangxi Bama mini-pig PGC-1αgene, and to analyze the expression of PGC-1αgene in various tissues of mini-pigs using RT-PCR and QRT-PCR techniques.Methods The PGC-1αgene coding sequence ( CDS) was amplified by PCR from the cDNA of longissimus muscle of Guangxi Bama mini-pig. The PCR products were inserted into pEASY-T5 vector, transfected E.coli, identified and sequenced.The PGC-1αgene expression in different tissues of the Bama mini-pigs was detected by RT-PCR and QRT-PCR assays.Results The PGC-1αgene CDS of Guangxi Bama mini-pig was cloned.It was 2391 bp in length.It had 99.9%homology with the reference sequence, and had two synonymous mutations that were C-A1105 and G-A1524.The expression level of PGC-1αgene was higher in the heart and kidney, followed by liver, subcutaneous fat and longissimus muscle, but the expression was not de-tected in pancreas of Guangxi Bama mini-pig.Conclusions We have successfully cloned the PGC-1αgene of Guangxi Bama mini-pig, and detected this gene expression in six tissues.The results of this study will provide a basis for studying the effect of PGC-1αon type 2 diabetes mellitus (T2DM) in Bama mini-pigs.

Objective To observe the influence on early blood glucose by 3 L fluid replacementafter kidney transplantation. Methods Patients(60 cases) after kidney transplantation were randomly di-vidod into two groups.Group A used circular fluid replacement and Group B used 3L fluid replacement. Thelevel of blood glucose was detected before operation,after operation ,once every 8 hours at the first day,thenonce a day for the following six days. Results The level of blood glucose with 3 L fluid replacement waslower than that with circular fluid replacement,especially from the fast day to the fourth day after operation(P < 0.05). Conclusions The method with 3 L fluid replacement surpasses obviously circular fluid re-placement in blood glucose control after kidney transplantation.

Objective By detecting the mutations spectrum of phenylalanine hydroxylase(PAH)gene in phe-nylketonuria(PKU)patients and their parents. The researchers analyzed the gene mutation features and high - frequency mutations and determined the relationship between the genotype and the phenotype,which would provide a theoretical basis for the early diagnosis and genetic consultation of PKU children in the region. Methods In this study,13 exons and their flanking introns of the PAH gene in 32 PKU patients and their parents from Wuxi and Suqian in Jiangsu province were sequenced by using the next - generation sequencing(NGS)technology. Results Sixty - one mutant sites and 32 mutant genes were detected in 32 PKU patients,and the mutation detection rate was 95. 31%(61 / 64 cases). The variants at c. 721C ﹥ T,c. 1068C ﹥ A,c. 611A ﹥ G,c. 1197A ﹥ T,c. 728G ﹥ A,c. 331C ﹥ T and c. 442 -1G ﹥ A were common mutations in the region with mutation frequency over 5% . What's more,4 novel variants of c. 699C ﹥ G,c. 265C ﹥ T,c. 722G ﹥ A and c. 1194A ﹥ G were found. Of those,c. 699C ﹥ G was not recorded in the PAH variant database and HGMD database and. c. 265C ﹥ T,c. 722G ﹥ A,and c. 1194A ﹥ G were first reported in the Chi-nese population. Genotype - accurate biochemical phenotype correlation by using the Guldberg AV system revealed con-sistency rate of 38. 0%(8 / 21 cases),which the consistency rate between accurate biochemical phenotype and predic-tive phenotype of moderate to severe genotype was 92. 3%(12 / 13 cases),and mild genotype was 50. 0%(4 / 8 cases). Conclusions The PAH gene variants of PKU patients in Jiangsu province are distributed mainly in exons 7,of which the highest frequency gene mutation is c. 721 c ﹥ T. Moreover,one novel variant c. 699C ﹥ G was reported for the first time. The PKU children inherit the PAH mutation gene mainly from both parents. There are definite correlation between the genotypes and phenotypes.

Objective To construct a nomogram to predict the prognosis of patients with gallbladder cancer(GBC).Methods The clinicopathological data of GBC patients were extracted from the SEER database,and the independent prognostic factors of GBC patients were analyzed by Cox regression,and a nomogram was constructed.Finally,the column diagrams in the training queue and validation queue are verified.Results Age,T stage,M stage,histological grade,radiotherapy,surgery and tumor size were independent prognostic factors in GBC patients,and the differences were statistically significant(P＜0.05).In the training cohort,the C index was 0.735(95％CI=0.721～0.749),and the AUC values at 1,3 and 5 years were 0.821,0.820 and 0.833,respectively.In the verification group,the C index was 0.733(95％CI=0.711～0.755),and the AUC values for 1,3 and 5 years were 0.816,0.807 and 0.827,respectively.The calibration curve shows that the predicted values of the nomogram are in good agreement with the observed values.The decision curve shows that the nomogram model has better prediction ability than TNM staging system.Conclusion The constructed dynamic prognosis nomogram of GBC patients has high accuracy and reliability.

Objective To study the application of walking combined with Buerger exercise in the treatment of grade 0 diabetic foot.Methods Eighty patients with grade 0 diabetic foot were randomly divided into observation group and control group.The control group was trained by walking exercise .The observation group was trained by walking combined with Buerger exercise through 12 months of training.The changes of self-sensory symptoms,ABI and DAWV were observed.Results The self-sensory symptoms of the two groups were significantly decreased (observation group: markedly effective 15 cases,effective 19 cases;control group: markedly effective 5 cases,effective 23 cases).The effective rate of the observation group was higher than that of the control group (85% vs.70%,χ2 =7.831,P <0.05).The ABI ratio of the two groups were higher than those before treatment ,and the ABI ratio of the observation group was higher than that of the control group [(0.889 ±0.113) vs.(0.842 ±0.124),t =1.772 P <0.05].The ratio of DAWV in the observation group was higher than that in the control group [(6.772 ±0.435)cm/s vs.(6.543 ±0.552)cm/s,t =2.061,P <0.05].Conclusion Walking combined with Buerger exercise can improve the blood flow velocity and blood flow of the collateral circulation of the lower limbs of patients with grade 0 diabetic foot,and improve the self-sensory symptoms of peripheral neuropathy .

Objective:To explore the predictive value of complement and coagulation indicators in sepsis related acute kidney injury (AKI).Methods:Clinical data of 217 patients with sepsis admitted to the Department of Internal Medicine and Intensive Care Unit of Affiliated Hospital of Jiangnan University from January 2018 to June 2019 were retrospectively analyzed. All patients were divided into sepsis with AKI group and without AKI group. Laboratory indicators of all patients were collected, including complement C 3, complement C 4, activated partial thrombin time (APTT), prothrombin time (PT), international normalized ratio (INR), D-dimer, procalcitonin(PCT), etc. logistic regression analysis was used to explore the risk factors of sepsis related AKI. Receiver operating characteristic curve (ROC) was used to evaluate the predictive value of independent risk factors. Results:Among 217 patients, 120 patients developed sepsis related AKI and 97 patients didn′t. PCT, lactic acid, PT, APTT, INR and D-dimer in AKI patients were significantly higher than those without AKI ( P<0.01). Complement C 3 and complement C 4 were significantly lower in AKI group ( P<0.01). Multivariate logistic regression analysis suggested that blood pressure<90/60 mmHg (1 mmHg=0.133 kPa)( OR=3.705, 95% CI 1.536-8.934, P=0.004), increased lactic acid ( OR=1.479, 95% CI 1.089-2.008, P=0.012), decreased complement C 3 ( OR=0.027, 95% CI 0.005-0.152, P<0.001) and prolonged APTT ( OR=1.090, 95% CI 1.047-1.137, P<0.001)were independent risk factors predicting AKI. The area under the ROC curve (AUC) of these multivariates were 0.741 (95% CI 0.675-0.807), 0.798 (95% CI 0.732-0.864), 0.712 (95% CI 0.643-0.781) and 0.716 (95% CI 0.648-0.783) respectively. The relevant sensitivity was 57.5%, 80.8%, 87.5%, 59.2%, and the specificity was 90.7%, 75.3%, 51.5%, 77.3%, respectively. The AUC of the combined four indicators was 0.880 (95 %CI 0.835-0.926) with the sensitivity 75.0% and the specificity 90.7%. Conclusion:The low level of complement C 3 and prolonged APTT predict sepsis related AKI, and the predictive value can be enhanced if hypotension and hyperlactacidemia are added.

Objective To investigate the effect ofDihuangyinzi (DHYZ)-medicated serum on receptor for advanced glycation end product (RAGE)/p38 miotgen-activated protein kinase (MAPK) /nuclear factor (NF)-κB pathway in SH-SY5Y cells induced by Aβ1-42.Methods Male SD rats were randomly divided into normal control group and experimental group (n=20);natural sera medium and DHYZ sera medium were prepared.(1) SH-SY5Y cells were divided into control group,model group and DHYZ treatment group;natural sera medium,natural sera medium+Aβ1-42 oligomer,and DHYZ sera medium+Aβ1-42 oligomer were given to the cells,respectively.Westem blotting was used to detect the protein expressions of NF-κB p65,p38 and phosphorylate (p)-p38.(2) SH-SY5Y cells were given DHYZ sera medium+Aβ1-42 oligomer treatment,and at different time points of Aβ1-42 oligomer treatment (15 min,30 min,60 min,12 h,24 h,48 h and 72 h),Western blotting was used to detect the protein expressions of p38 and p-p38.(3) SH-SY5Y cells were divided into 6 groups:mock-transfected RAGE blank group,transfected RAGE blank group,mock-transfected RAGE model group,transfected RAGE model group,mock-transfected RAGE herb group and transfected RAGE herb group;herb groups were given DHYZ-medicated serum;inflammatory factors,interleukin (IL)-1β,IL-6,and tumor necrosis factor (TNF)-a,were measured by ELISA and cytometric bead array.Results (1) As compared with model group,DHYZ treatment group had significantly decreased NF-κB p65 and p-p38/p38 protein expression.(2) The p-p38 protein expression began to increase 30 min after Aβ1-42 treatment,reached to its peak level 24 h after Aβ1-42 treatment,and began to decrease 48 h after Aβ1-42 treatment.(3) The IL-1β,IL-6 and TNF-α levels were increased significantly in the transfected RAGE model group as compared with those in the mock-transfected RAGE model group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were increased significantly in the transfected RAGE herb group as compared with those in the mock-transfected RAGE herb group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were decreased significantly in the mock-transfected RAGE herb group as compared with those in the mock-transfected RAGE model group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were decreased significantly in the transfected RAGE herb group as compared with those in the transfected RAGE model group (P＜0.05).Conclusion DHYZ-medicated serum could inhibit the RAGE-p38 pathway and improve the inflammatory reaction in Aβ1-42-induced SH-SY5Ycells transfected with RAGE gene to protect the SH-SY5Y cells.

Objective:To investigate the effects of extracorporeal carbon dioxide removal (ECCO 2R) combined with continuous renal replacement therapy (CRRT) on respiratory efficiency and diaphragm function in patients with acute respiratory distress syndrome (ARDS) received mechanical ventilation. Methods:A prospective randomized controlled study was conducted. Sixty patients with mild to moderate ARDS admitted to the department of respiratory and critical care medicine of Henan Provincial People's Hospital from January 2019 to January 2021 were enrolled, and they were divided into observation group and control group according to the random number table method, with 30 cases in each group. All patients received antibiotics, anti-inflammatory, and mechanical ventilation therapy. On this basis, the observation group received ECCO 2R and CRRT, while the control group received bedside CRRT. Baseline data including gender, age, etiology, acute physiology and chronic health evaluationⅡ(APACHEⅡ), etc., were recorded. Arterial blood gas analysis [including arterial partial pressure of oxygen (PaO 2), arterial partial pressure of carbon dioxide (PaCO 2), and oxygenation index (PaO 2/FiO 2)] was performed at 12 hours and 24 hours during the treatment, and respiratory mechanics parameters [including tidal volume, respiratory rate, maximum expiratory pressure (MEP), and maximum inspiratory pressure (MIP)] were recorded, and rapid shallow breathing index (RSBI) was calculated. The levels of glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and superoxide dismutase (SOD) in serum were detected by enzyme-linked immunosorbent assay (ELISA). Diaphragm thickness and diaphragm activity were measured by ultrasonography at 24 hours during the treatment. Results:There were no significantly differences in age, gender, etiology, and APACHEⅡ score between the two groups, indicating that the baseline data of the two groups were balanced and comparable. Compared with the 12 hours after treatment, the PaO 2 and PaO 2/FiO 2 in the observation group significantly increased, PaCO 2 significantly decreased, RSBI significantly decreased, MEP and MIP significantly increased, and serum GSH-Px and MDA significantly decreased, while SOD significantly increased at 24 hours during the treatment. In the control group, only PaCO 2 significantly decreased. Compared with the control group, the PaCO 2 significantly decreased in the observation group at 12 hours and 24 hours [mmHg (1 mmHg≈0.133 kPa): 55.05±7.57 vs. 59.49±6.95, 52.77±7.88 vs. 58.25±6.92, both P < 0.05], but no significantly differences in PaO 2 and PaO 2/FiO 2. Compared with the control group, the observation group showed significant decreases in RSBI at 12 hours and 24 hours (times·min -1·L -1: 85.92±8.83 vs. 90.38±3.78, 75.73±3.86 vs. 90.05±3.66, both P < 0.05), significant increases in MEP and MIP [MEP (mmH 2O, 1 mmH 2O≈0.01 kPa): 86.64±5.99 vs. 83.88±4.18, 93.70±5.59 vs. 85.04±3.73; MIP (mmH 2O): 44.19±6.66 vs. 41.17±3.13, 57.52±5.28 vs. 42.34±5.39, all P < 0.05], and significant decreases in serum GSH-Px and MDA [GSH-Px (mg/L): 78.52±8.72 vs. 82.10±3.37, 57.11±4.67 vs. 81.17±5.13; MDA (μmol/L): 7.84±1.97 vs. 8.71±0.83, 3.67±0.78 vs. 8.41±1.09, all P < 0.05], as well as a significant increase in SOD (U/L: 681.85±49.24 vs. 659.40±26.47, 782.32±40.56 vs. 676.65±51.97, both P < 0.05). Compared with the control group, the observation group showed significant increases in diaphragm thickness and diaphragm activity at 24 hours of treatment [diaphragm thickness (cm): 1.93±0.28 vs. 1.40±0.24, diaphragmatic thickening fraction: (0.22±0.04)% vs. (0.19±0.02)%, quiet breathing diaphragm displacement (cm): 1.42±0.13 vs. 1.36±0.06, deep breathing diaphragm displacement (cm): 5.11±0.75 vs. 2.64±0.59, all P < 0.05]. Conclusion:ECCO 2R combined with CRRT can reduce work of breathing and oxidative stress levels in ARDS patients receiving non-invasive ventilation, and protect diaphragm function.