Objective To investigate effect of artesunate on expressions of Toll-like receotor-4 (TLR4) and interleu?kin-8 (IL-8) in renal tissue of diabetic nephropathy rats. Methods Male SD rats of over 200 g in body weight (n=45) were injected with low dosage streptozotocin and fed with high fat and sucrose diets to establish diabetes nephropathy (DN) model. Among all rats of successful model, thirty-two were randomly selected and divided into four groups (n=8 in each group), Mod?el group (Group B), Artesunate at high dose treatment group [30 mg/(kg · d)] (Group C), Artesunate at low dose treatment group [10 mg/(kg·d)] (Group D), Enalapril treatment group [10 mg/(kg·d)] (Group E). Another eight rats without STZ injec?tion whose body weight is under 200 g were assigned as Normal group (Group A). Then, rats in Group A and B were given the same volume of normal saline [10 mg/(kg·d)] while rats in Group C, D and E were given 30 mg/(kg·d) Artesunate, 10 mg/(kg·d) Artesunate and 10 mg/(kg · d) Enalapril respectively intragastrically for consecutive 12 weeks. Fasting blood glucose, body weight, kidney index, 24-hours proteinuria, serum creatinine (Cr) and blood urea nitrogen (BUN) were measured. Expression level of TLR4 in renal tissue of rats were examined by Western blot;ELISA was used to quantify the concentration of IL-8 in serum and renal tissue of rats. Results Compared with rats in Group A, the levels of Fasting blood glucose, kidney index , 24-hours proteinuria, Cr and BUN as well as the level of TLR4 in kidney, levels of IL-8 in serum and kidney all significant?ly increased in rats in Group B, C, D and E(P<0.05). On the other hand, body weight were decreased in rats of Group B, C, D and E compared to rats in Group A(P<0.05). The level of TLR4 in kidney, levels of IL-8 in serum and kidney, 24-hours proteinuria, Cr and BUN in rats of Group C, D and E were significantly lower than those in rats of Group B(P<0.05). Furthermore, compared with rats in Group D, the levels of TLR4 in kidney, IL-8 in serum and kidney, 24-hours proteinuria, Cr and BUN were decreased in rats in Group C and E(P<0.05). Pearson correlation analysis showed that the expression lev?el of TLR4 positively correlated with IL-8 level in serum (r=0.969, P<0.05), IL-8 level in kidney (r=0.972, P<0.05), 24-hours proteinuria(r=0.965, P<0.05), Cr(r=0.944, P<0.05)and BUN(r=0.903, P<0.05). IL-8 level in kidney positively correlated with 24-hours proteinuria(r=0.962, P<0.05), Cr(r=0.929, P<0.05)and BUN(r=0.940, P<0.05). Conclu?sion Artesunate decreases expression of TLR4 and IL-8, reduce 24-hours proteinuria, inhibits the inflammation of the DN,relives the pathological lesions of nephron rats with diabetic nephropathy.

Objective To investigate the effects of artesunate( Art) on renal function and the expression of monocyte chemotactic protein-1(MCP-1)and tumor necrosis factor-α(TNF-α)in rats with type 2 diabetic nephropathy(DN). Methods Thirty six male SD rats were randomly divided into six groups, six rats in each, as the normal control, model control, low-dose Art, middle-dose Art, high-dose Art groups which were intragastrically administrated with Art at the dose of 10, 20 and 30 mg·kg-1 ·d-1 for consecutive 8 weeks, enalapril group, which were intragastrically administrated with enalapril 10 mg·kg-1 ·d-1 for consecutive 8 weeks. Rats were injected with low dosage streptozotocin and fed with high fat and high sugar diets to establish type 2 diabetic nephropathy rat model. After 8 weeks’ treatment, the plasma glucose, 24-hour urine protein, serum creatinine and blood urea nitrogen were measured to evaluate renal function. The pathological morphology of rats was performed. Immunohistochemistry and enzyme linked immunosorbent assay ( ELISA ) were performed to examine the expression levels of MCP-1 and TNF-α,respectively. Results Compared with the model control group, 24-hour urine protein, serum creatinine, blood urea nitrogen and blood glucose were significantly decreased in the treatment groups (P<0. 01). Art significantly decreased the serum expressions of MCP-1 and TNF-α in low-, middle-, high-dose Art group ( 157. 47 ± 38. 53, 138.65±18.03,105.09±12. 64 and 3. 14±0. 38,2. 58±0. 11,2. 25±0. 16) pg·mL-1, compared with model control group (181.71±23.06 and 3. 98±0. 24) pg·mL-1(P<0. 05). Conclusion Art has beneficial effects on type 2 diabetic nephropathy. The mechanism of which may be related to the inhibition of inflammatory factor MCP-1 and TNF-α,further relive the pathological injury of kidney and delay the progress of diabetic nephropathy to some extent.

Objective To investigate the short-term clinical efficacy and complication of esophageal cancer patients with two-field lymph node dissection by thoracolaparoscopic esophagectomy surgery and open surgery. Methods One hundred and fifty esophageal cancer patients with two-field lymph node dissection were selected, and they were divided into control group (using open surgery, 75 cases) and observation group (using thoracolaparoscopic esophagectomy surgery, 75 cases) by random digits table method. The operation time, bleeding amount, hospital staying time, number of lymph node dissection, reoperation rate, intensive care unit (ICU) transferring rate and postoperative complication were compared. Results There was no statistical difference in operation time between 2 groups ( P>0.05). The bleeding amount and hospital staying time in observation group were significantly lower than those in control group: (210.33 ± 30.71) ml vs. (254.59±35.28) ml and (8.45±1.52) d vs. (11.61±2.08) d, there were statistical differences (P<0.05). The number of chest lymph node dissection in observation group were significantly higher than those in control group:(17.20±4.06) pieces vs. (10.44±2.65) pieces, and there was statistical difference (P<0.05). There were no statistical differences in reoperation rate and ICU transferring rate between 2 groups ( P>0.05). There were no statistical differences in incidences of hoarseness and anastomotic stenosis between 2 groups ( P>0.05). The incidences of pulmonary infection and arrhythmia in observation group were significantly lower than those in control group:17.33%(13/75) vs. 30.67%(23/75) and 2.67%(2/75) vs. 14.67%(11/75), and there were statistical differences (P<0.05). Conclusion Compared with open surgery, thoracolaparoscopic esophagectomy surgery with two-field lymph node dissection for esophageal cancer patients can effectively reduce the degree of operative trauma, accelerate postoperative rehabilitation process, improve the effects of lymph node dissection, and reduce postoperative complication risk.

Objective:To observe the effect of liver-soothing and mind-regulating acupuncture method on the resting-state electroencephalography (EEG) in rats with post-traumatic stress disorder (PTSD),and to provide evidence for the effect mechanism study and clinical application of acupuncture intervention for PTSD.Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group,a model group,a grasping group,a paroxetine group and an acupuncture group,with 12 rats in each group.Except for rats in the blank control group,rats in the other groups were subjected to preparing the PTSD models using 'incarceration plus electric shock' method.After interventions,changes in rat behavior of each group were observed;changes in resting-state EEG were collected and analyzed with multichannel EEG acquisition and analysis system,and image analysis and statistical processing were performed.Results:Compared with the blank control group,the average escape latency in the model group was significantly longer (P＜0.05),and the times of crossing the platform and the effective areas were all significantly reduced (P＜0.01).Compared with the grasping group,the average escape latencies in the paroxetine group and acupuncture group were significantly shortened (P＜0.05),and the times of crossing the platform and the effective areas were all significantly increased (P＜0.05).There were no significant differences in the average escape latency,the times of crossing the platform and the effective areas between the acupuncture group and paroxetine group (all P＞0.05).Compared with the blank control group,the α-wave power spectrum value in the model group was significantly decreased,and the power spectrum values of β-wave,δ-wave and a-wave were significantly increased (all P＜0.01);compared with the grasping group,α-wave power spectrum values in the paroxetine group and acupuncture group were significantly increased (both P＜0.01),and the power spectrum values of β-wave,δ-wave and a-wave were decreased significantly (all P＜0.01).The power spectrum values of α-wave,β-wave,δ-wave and (e)-wave of rats in the acupuncture group were not significantly different from those in the paroxetine group (all P＞0.05).Conclusion:Liver-soothing and mind-regulating acupuncture method can significantly improve the abnormal EEG activity in PTSD rats,which may be one mechanism of liver-soothing and mind-regulating acupuncture method in effectively affecting the brain function in PTSD rats.

BACKGROUND:An effective freezing-thawing technique is crucial for the clinical application of human umbilical cord mesenchymal stem cells(UC-MSCs).OBJECTIVE:To investigate biological characteristics of UC-MSCs after cryopreservation.METHODS:UC-MSCs were isolated from human umbilical cord and frozen in liquid nitrogen.The survival rate and the suppressive effect of γ-interferon(IFN-γ)of cryopreserved-thawed and fresh human UC-MSCs were compared.Furthermore,the multiple potentials and phenotype of UC-MSCs were estimated after cryopreservation.RESULTS AND CONCLUSION:There was no significant difference between cryopreserved-thawed and fresh human UC-MSCs on the survival rate and the suppressive effect of IFN-γ of peripheral blood mononuclear cells(PBMCs).After cryopreservation,human UC-MSCs had the potential differentiation and the phenotype of mesenchymal stem cells.

Objective:To observe the effect of acupuncture on blood oxygen concentration in the brain of rats with post-traumatic stress disorder (PTSD) based on functional near-infrared spectroscopy (fNIRS),thus to reveal the mechanisms of acupuncture in intervening the brain function of PTSD rats.Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank group,a model group,a grasping group,a paroxetine group and an acupuncture group,with 12 rats in each group.Except the blank group,rats in the other groups all received incarceration plus electric shock for 7 d to prepare the PTSD animal model.One hour before the stress model was established,rats in each group received the designated intervention:rats in the blank group and the model group did not receive any intervention;rats in the grasping group received grasping and fixation;rats in the paroxetine group received paroxetine hydrochloride solution by intragastric administration;and rats in the acupuncture group received acupuncture.Six-day treatment was a course,with 2 courses of treatment conducted for a total of 12 d.After the modeling,rats in each treatment group received intervention for 5 d,and the fNIRS system was used to collect and record the changes in the concentrations of oxygenated hemoglobin (HbO2),deoxygenated hemoglobin (d-Hb) and total hemoglobin (t-Hb) of the involved rat's brain regions,and also to assess the brain function.Results:Compared with the blank group,the concentration of HbO2 was significantly increased,the concentration of d-Hb was significantly decreased,and the concentration of t-Hb was significantly increased in the model group and the grasping group after the intervention,and the differences were statistically significant (all P＜0.01).Compared with the model group,the concentrations of HbO2,d-Hb and t-Hb in the grasping group did not change significantly (all P＞0.05).Compared with the grasping group,the concentration of HbO2 was significantly decreased,the concentration of d-Hb was significantly increased,and the concentration of t-Hb was significantly decreased in the paroxetine group and the acupuncture group,and the differences were statistically significant (all P＜0.05).There were no significant differences in the concentrations of HbO2,d-Hb and t-Hb between the paroxetine group and the acupuncture group (all P＞0.05).Conclusion:Acupuncture can regulate the blood oxygen concentration in the brain of PTSD model rats,which may be an important mechanism of acupuncture in intervening the brain function in PTSD rats.

Objective To detect the expressions of CD70 mRNA and protein and to determine the methylation status of CD70 gene promoter in T cells from patients with systemic lupus erythematosus (SLE).Methods Peripheral blood CD4+ and CD8+ T cells were isolated from 15 patients with active SLE,15 patients with inactive SLE and 15 healthy control subjects.Real-time quantitative reverse transcription-PCR was carried out to quantify the mRNA expression of CD70,flow cytometry to determine the frequency of CD4+CD70+ and CD8+ CD70+ T cells,and bisulfite sequencing to evaluate the methylation status of CD70 gene promoter in CD4+ and CD8+ T cells.Differences in these parameters among these groups were analyzed by one-factor analysis of variance and SNK-q test.Results Compared with the healthy controls,the patients with active SLE and inactive SLE showed a significant increase in CD70 mRNA expression in CD4+ T cells (0.82 ± 0.12 and 0.73 ± 0.11 vs.0.45 ±0.09,F =53.017,P ＜ 0.01) and in the frequency of CD70+CD4+ T cells (80.30％ ± 11.04％ and 66.80％ ± 3.98％ vs.12.48％ ± 3.45％,F =311.517,P ＜ 0.01).Also,the expression of CD70 mRNA in CD4+ T cells and the frequency of CD70+CD4+ T cells were significantly higher in patients with active SLE than in patients with inactive SLE (both P ＜ 0.05).There was a positive correlation between the frequency of peripheral CD70+CD4+ T cells and disease activity in SLE in these patients (r =0.792,P ＜ 0.01).The average methylation index of the region between-600 bp and-300 bp of CD70 gene promoter in CD4+ T cells was 0.32 ± 0.05 and 0.36 ± 0.05 respectively in the patients with active and inactive SLE,significantly lower than that in the healthy controls (0.62 ± 0.05,F =152.64,P ＜ 0.01),and the patients with active SLE showed a significantly lower methylation index than those with inactive SLE (P ＜ 0.05).Conclusions The CD70 gene promoter in CD4+ T cells is significantly hypomethylated in patients with SLE,which may directly lead to the overexpression of CD70.

A series of novel sorafenib analogues were designed and synthesized. The cytotoxic activities of these compounds were tested in four tumor cell lines. Some of the compounds showed potent antiproliferative activity against the tested cell lines with IC50 = 4-20 micromol x L(-1). Some compounds demonstrated competitive antiproliferative activities to sorafenib against tested cancer cell lines. Among them, compound 7c demonstrated significant inhibitory activities on ACHN, HCT116 and MDA-MB-231 cell lines with IC50 values of 9.01, 4.97, 6.61 micromol x L(-1), respectively.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Inhibitory Concentration 50 ; Molecular Structure ; Niacinamide ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Phenylurea Compounds ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship

Single clones of Leptospirillum ferrooxidans were obtained on bilayer solid medium plate by incorporating Het-erotroph Acidiphilium SJH into the underlayer broth medium. The morphologies of the bacteria were investigated using electronic microscope.

Polymerase chain reaction was employed to amplify the whole HBV CP region from the serum of patients with chronic hepatitis B virus(HBV) infection, and then the PCR products were subcloned into pGEM Teasy vectors. Clones were randomly selected to be sequenced and the selected clones were compared to look for the difference.The sequencing results suggested that each sequence of selected clones was different and there were HBV quasispecies groups in patients. There were hot deletion region and point mutation near the TATA like box of CP gene. To address whether the mutations were responsible for the transcriptional activity, the wild type(wt) and the mutants of HBV CP genes were subcloned into pcDNA3 1( ) vectors, respectively. The reverse oriented clones were digested with KpnI and XhoI, and cloned into the KpnI and XhoI sites of the chloramphenicol acetyltransferase (CAT) expressing vector (pCAT3 basic).The recombinant CAT plasmids were transfected into HepG2 cells using lipofectamine PLUS reagent, and the CAT expression which indirectly represented the transcriptional activity of HBV CP lying upstream of CAT gene was detected with a CAT ELISA kit. The restriction enzyme digesting results indicated that the recombinant CAT plasmids were successfully constructed, and the transfection tests indicated that the transcriptional activity of the mutants with deletion or substitute point mutation of TATA like box were reduced in comparison with that of CPwt. The HBV CP gene heterogeneity downregulated the transcriptional activity to some extent.