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 effect of losartan on tubulointerstitial fibrosis of rat unilateral ureteral obstruction(UUO) model, and to study whether the use of large dose could be better exert a superior renoprotective effect than conventional dose. Methods Three days after UUO, rats were randomly assigned to conventional dose and large dose of losartan group, shamoperated group and operation group. Two treatment groups were administered orally with a daily dose of losartan 50 mg/kg or 500 mg/kg by gastric gavage. Sham-operated group and operation group received the same volume of physiological saline. Rats were sacrificed 7, 14, 21 days after obstruction. Daily urinary albumin excretion(UalbV), tail-cuff pressure(TCP), the percentage of renal tubular lesions, fractional interstitial area (INT), macrophage infiltration and the expression of transforming growth factor (TGF)-β1 mRNA were assessed and compared. Results The TCP, UalbV, percentage of tubular lesions and interstitial fibrosis, the number of interstitial macrophages and the expression of TGF-β1 mRNA were significantly increased in operation group as compared with other groups (P<0.05). Furthermore, in comparison with conventional dose, the large dose treatment significantly reduced TCP and UalbV, attenuated interstitial fibrosis and tubular lesions, suppressed macrophages infiltration and the expression of TGF-β1 mRNA. Conclusion The large dose of losartan provides superior renoprotection compared to the treatment with conventional dose in UUO model, which interrupts the positive feedback involved in the vicious cycle between inflammatory cell and Ang Ⅱ.

Objective To study the influence of bFGF gene transfected bone marrow-derived mesenchymal stem cells (BMSCs) on the inflammatory cytokines of COPD rat. Methods The BMSCs were separated from SD rat and cultured and then bFGF gene was imported to BMSCs by liposome transfection method. The samples were prepared into six groups: normal control group, COPD group (A), BMSCs group (B), pcDNA3.1-BMSCs group (C), bFGF-pcDNA3.1-BMSCs group (D), and bFGF group (E). The expressions of TNF-α and IL-1β by QRT-PCR were detected. Results Compared with COPD group, TNF-α and IL-1β genes from groups B to D dropped significantly (P < 0.05). The changes of TNF-α and IL-1β genes among groups B to D showed no significant difference (P > 0.05). Conclusion BFGF transfected BMSCs, sample BMSCs and pcDNA3.1 transfected BMSCs can inhibit the expression of inflammatory cytokines of TNF-α and IL-1β, but there is no obvious advantage in comparison to bFGF transfected BMSCs and sample BMSCs in respect of inhibiting the expression of inflammatory cytokines of TNF-α and IL-1β.

Objective To study the related factors of intradiscal injection of collagenase. Methods One hundred and sixty eight patients with lumbar disc herniation were injected with collagenase(600U) under X ray monitoring. Assessment of efficacy of the treatment was made in 1 week, 3 months and 1 year follow up. The parameters such as the size, the type and the number of herniated disc were studied. The effect of stenosis was also analyzed. Results The overall success rate after 1 week, 3 months and 1 year following the therapy was 22.62%, 89.29% and 92.26% , respectively. Seven patients were treated with surgical operation during the follow up, 1 developed cauda equina syndromes at the third day after the injection. It was found that the size of herniation did not affect the efficacy of the treatment. The protruded disc got better result than bulge disc after 1 week following the chemonucleolysis ( P

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.

BACKGROUND:Stable and efficient labeling of dental pulp stem cel s in vitro is most important in tracer technique, which is also the basis of tooth regeneration in vivo.
OBJECTIVE:To determine the optimal condition and method for transfection of stem cel s derived from rat dental pulp with green fluorescent protein infection by lentiviral vector and to determine whether green fluorescent protein-labeled dental pulp stem cel s maintain their stem cel properties.
METHODS:Rat dental pulp stem cel s were obtained by modified enzyme digestion method, to identify the immune phenotype and differentiation potential. Dental pulp stem cel s were infected with green fluorescent protein by lentiviral vector for 24 and 48 hours at different multiplicity of infection (MOI) (5, 10, 25, 50 and 100). The infection efficiency and fluorescence intensity were analyzed by inverted fluorescent microscopy. The clonal and proliferation ability, cel cycle and the mineralization potential were compared before and after transfection. Based on those mentioned above, we could evaluate the influence of infection on their biological characteristics.
RESULTS AND CONCLUSION:Flow cytometry results showed that rat dental pulp stem cel s expressed STRO-1 and CD146 but not CD34 or CD45. The dental pulp stem cel s could differentiate into osteoblasts and
adipocytes when cultured in specific medium for each lineage differentiation. The highest efficiency of infection and strongest fluorescence expression appeared at 48 hours of infection and MOI 50. There were no significant differences in growth ability, cel colony formation rate and cel cycle before and after transfection (P>0.05). And the alkaline phosphatase expressed positively. Infection for 48 hours at MOI 50 is optimal for transfecting dental pulp stem cel s with green fluorescent protein by a lentiviral vector, thereby providing reliable tracer method for the study of rat dental pulp stem cel s in vivo.

Objective To investigate the effects of artesunate (Art) on cell apoptosis, tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8) expression induced by high glucose in rat renal tubular epithelial cells (NRK-52E). Methods NRK-52E cells were cultured and divided into normal control group, high glucose group, high glucose with different concen?trations of Art (10 mg/L, 20 mg/L and 30 mg/L) groups, and high glucose with Ena (5 mg/L) group. MTT assay was used to de?tect the cell proliferation. The apoptotic rate was evaluated by flow cytometry with AnnexinV-FITC/PI double stains. The pro?tein levels of TNF-αand IL-8 in the cell culture supernatant were determined using ELISA. Results High glucose inhibit?ed NRK-52E proliferation, induced its apoptosis, and the expressions of TNF-αand IL-8 in the supernatant. Application of Art obviously abolished the effects of high glucose, and the effects of Art were showed in dose-dependent manner. Conclu?sion Art can suppress high glucose-stimulated cell apoptosis, enhance TNF-αand IL-8 expression in NRK-52E cells. The anti-inflammatory action and immune regulation of Art could be a novel approach of treating diabetic nephropathy.

Objective:To investigate the clinical features of IgD multiple myeloma (MM) and the effect and prognosis of daratumumab-based combination therapy.Methods:The clinicopathological data of a IgD MM patient with disease progression and extramedullary infiltration treated with daratumumab in the Affiliated Hospital of Qingdao University in December 2019 were retrospectively analyzed.Results:The 74-year-old woman was diagnosed as IgD MM by bone marrow aspiration and immunofixation electrophoresis. The patient was given VD (bortezomib, dexamethasone), RD (lenalidomide, dexamethasone) and ID (ixazomib, dexamethasone) regimens. In June 2020, the patient developed multiple subcutaneous nodules, and she was assessed as progressive disease with extensive extramedullary infiltration. After treated with daratumumab-PAD (liposomal doxorubicin, bortezomib, dexamethasone) regimen, the patient's subcutaneous nodules were significantly reduced and partially disappeared, and the general condition was significantly improved. But the patient was in a cachexia state and finally died of the irregular treatment and disease progression.Conclusions:IgD MM has a low incidence and a short survival period, and there is no uniform standard treatment. The early application of daratumumab combined with proteasome inhibitors, immunomodulators, cytotoxic drugs and hematopoietic stem cell transplantation may improve the overall survival of patients.

Objective To evaluate the potential malignancy, prognosis and risk factors for intraductal papillary mucinous neoplasm(IPMN), which were classified into different risk levels based on Fukuoka guideline. Methods A retrospective analysis of patients with IPMN diagnosed at Nanjing Drum Tower Hospital from 2009 to 2016 was conducted. Clinical characteristics,treatment and prognosis of IPMNs were analyzed. Results A total of 94 IPMN patients were included and divided into 3 groups according to Fukuoka guideline,46 patients in high-risk(HR)group,30 in group of worrisome features(WF), and 18 in low-risk(LR)group. For patients undergoing surgery treatment, there were 5 cases(19.2%,5/26)in HR group and 2 cases(12.5%,2/16)in WF group whose postoperative pathological findings were malignant (P=0.690). The 5-year survival rates after operations were 73.9% and 77.0% in HR and WF group, respectively(P=0.830). For patients without surgery treatment, in a 5-year follow-up, there were 6 cases (33.3%,6/18),2 cases(16.7%,2/12)and 0(0.0%,0/18)progressing into pancreatic cancers in HR, WF and LR groups,respectively(P<0.05). In addition,among the three groups,the 5-year survival rates were 49.5%,85.7% and 100.0%(P=0.025). Jaundice was significantly related to prognosis(P<0.01) and the hazard ratio was 8.883(95%CI:2.953-26.721). Conclusion Jaundice is a predictive risk factor for survival of IPMN. As for the treatment to IPMN, patients in HR group should receive surgery treatment while those in LR group can be followed up. For patients in WF group,the treatment should be customized, with evaluation of predictive risk factors,and operations can be performed when needed.