1.Treatment of diabetic gastroparesis: current status
Chinese Journal of General Practitioners 2021;20(6):700-704
Diabetic gastroparesis (DGP) is a common complication in patients with diabetes mellitus of long duration, presenting with recurrent nausea, vomiting, postprandial fullness and early satiety caused by delayed gastric emptying. The treatments of DGP include dietary therapy, nutritional support, glycemic control, use of prokinetic and antiemetic agents. This review focuses on current status of the drug treatment and the progress of new agents of DGF under the preclinical and clinical trials.
2.Low-density lipoprotein promotes epithelial-mesenchymal transition and extracellular matrix accumulation in human peritoneal mesothelial cells
Yanhui FANG ; Lanping JIANG ; Limeng CHEN ; Xuemei LI ; Xuewang LI
Chinese Journal of Nephrology 2013;(1):44-49
Objective To investingate the effect of low-density lipoprotein (LDL) on epithelial -mesenchymal transition and extracellular matrix (ECM) accumulation in human peritoneal mesothelial cells (HPMCs).Methods (1)HPMCs were randomly divided into control group,LDL group (100 mg/L) and LDL (100 mg/L) + lactoferrin (100 mg/L,LDL receptor blocking agent) group.After co-cultured for 24 h,the expression of LDL receptor in HPMCs was examined by immunofluorescence staining,and the LDL uptake by HPMCs was observed with oil red O staining.(2)HPMCs were cultured with different concentrations of LDL (0,25,50,100 mg/L).After co-cultured for 24 h,the change of cell morphology was observed by inverted phase contrast microscope,and the expression of α-smooth muscle actin (α-SMA) was examined by immunofluorescence.(3) HPMCs were randomly divided into control group (5.6 mmol/L glucose),mannitol group (M,2.18% mannitol),low glucose group (LG,30 mmol/L),high glucose group (HG,120 mmol/L) and HG + LDL group (120 mmol/L glucose + 100 mg/L LDL).Cocultured for 48 h,the mRNA expression of α-SMA,E-cadherin and type 1 plasminogen activator inhibitor (PAI-1) was detected by real-time quantitative PCR,the protein expression of α-SMA was detected by Western blotting,the content of type I collagen (Col I) and PAI-1 in supernatant was detected by ELISA.Results (1) After co-cultured with LDL for 24 h,the expressin of LDL receptor was found on the cell membrane of HPMCs.Oil red staining showed that LDL could be uptaken into the cells and abolished by LDL receptor blocker.(2) HPMCs tended to be loosely intercellular connected to each ofher,and prsesnted significant formation of fibroblast-like spindle morphology.The cytoplasm immunofluorescence intensity of α-SMA gradually increased with the increase of LDL concentration.Compared to the control group,the expressions of α-SMA mRNA and protein were significantly increased,and the expression of E-cadherin mRNA was decreased in HG + LDL group(all P < 0.05).But the expressions of the parameters above-mentioned were not significant different between HG group and HG + LDL group or between HG group and control group.(3) Compared with HG group or control group,the concentrations of Col Ⅰ [(19.27±0.17) μg/L vs (14.09±0.30) μg/L or (14.81±0.91) μg/L,all P < 0.05] and PAI-1 [(498.24±76.91) ng/L vs (342.19±30.43) ng/L or (220.39±33.82) ng/L,all P < 0.05] in supernatant of HPMCs were significantly up-regulated in HG + LDL group,meanwhile the expression of PAI-1 mRNA was significantly higer than that in control group (P =0.022).Conclusions HPMCs uptake LDL into cells via LDL receptors.LDL can induce HPMCs transdifferentiation in the condition of high glucose,increase the secretion of Col Ⅰ,inhibit the degradation of ECM through up-regulating the expression of PAI-1,and lead to ECM accumulation.
3.Effects of nerve growth factor on the neural cells and Egr-1 gene expression in the brain of childhood intractable epileptics
Qiong FANG ; Yanhui CHEN ; Pengfan YANG ; Min LU
Journal of Clinical Pediatrics 2009;(11):1019-1022,1035
Objective To investigate the effect of nerve growth factor (NGF) on the neural cells and expression of early growth response gene-1 (Egr-1 gene) in the hippocampus discharge zone of childhood intractable epileptics. Methods Acutely dissociated cell suspensions of childhood hippocampus discharge zone with non specific pathological changes (n = 16) were exposed to NGF group and control group respectively. There were three subgroups exposed to NGF at the levels of 12.5, 50, 100 ng/ml in NGF group. After 4 hr culturing, the astrocytes and neurons were lablled by Bb immunostain with the specific markers such as GFAP and MAP2. The total number of neural cells was counted under inversion fluorescence microscope and the Egr-1mRNA expression was detected by semiquantitative RT-PCR analysis. Results There were significant differences of the numbers of neural cells survived in hippoeampus region between the NGF group and the non-added NGF group (P < 0.01). Among the different levels of NGF 12.5, 50, 100 ng/ml, the number of the total cultural neurons and GFAP(+) astrocytes, MAP_2(+) neurons were increased with ascending levels of NGF (P < 0.05). At the same time, the expression of Egr-1mRNA (A_(Egr-1)/A_(β-actin)) in the NGF groups was also increased (P < 0.01). There was positive correlation between the number of the survivable neural cells and Egr-1mRNA quantity (r = 0.780, P < 0.01) among the three NGF groups. Conclusions NGF can protect the neural cells of epileptic discharge zone by promoting the expression of Egr-1 gene.
4.Period of Validity for Skin Disinfectant Povidone-iodine
Fang WANG ; Zanmei LU ; Yanhui LIU ; Lihua ZHANG ; Meijuan DENG
Chinese Journal of Nosocomiology 2006;0(05):-
OBJECTIVE To study the period of validity of iodine skin disinfectant povidone-iodine(Iodophor) in different seasons,different hours and different using frequency.METHODS In spring and summer respectively,the iodine was used for 10 days continually after the bottle opened.Then,the sample drawing from bottle was detected for iodine content,pathogenic bacteria and contamination of bacteria quantification.RESULTS The available iodine content was coming down gradually with the increasing in usage time and frequency.And the changing in the content differed in spring and summer.Microorganism was not found in povidone-iodine under using time,different seasons and using frequency.CONCLUSIONS The skin disinfectant can be used for 10 days at least under standard range after begining using.
5.Changes of A20 expression in mesangial cells of LPS-induced diabetic nephropathy rat model
Yaling LIANG ; Jiao CHEN ; Yang LONG ; Yanhui LI ; Qiuhan LI ; Fang FAN ; Yong XU
Tianjin Medical Journal 2016;44(4):444-448
Objective To observe the changes of A20 in mesangial cells of diabetic nephropathy (DN) rat model in?duced by lipopolysaccharide (LPS)-rat, and to explore its possible mechanism. Methods (1)Thirty health male Wistar rats were randomly divided into two group. Model rats were given streptozotocin (STZ) at a dose of 60 mg/kg by intraperitoneal in?jection. Rats in the control group received the same volume of citrate buffer in the same way. Levels of blood glucose and uri?nary microalbumin were detected in two groups at the 6th and the 8th week. Changes of renal pathology were observed by HE staining. Changes of protein A20 were observed by immunohistochemistry. (2) Expression changes of gene and proteins A20, nuclear factor (NF)-κB, IκB, IKKγand MCP-1 in renal cells treated with LPS were determined after treatment with different time points (0, 2, 4, 6, 12, 24, 48 and 72 h) and different concentrations (0.1, 1 and 10μg/L). Results (1) Levels of blood glucose and urinary microalbumin were significantly increased in model group compared with those of control group ( P <0.01). HE stainig showed that hyaline degeneration in tubular epithelial cells was found in model group, especially at the 8th week. Results of immunohistochemistry showed that expression of protein A20 significantly decreased in kidney tubules and nearly disappeared in glomerulus in model group compared with that of control group, which expressed less at the 8th week. (2) There was no significant difference in the expression of IKKγbetween different concentrations and different times. Com?pared with 0 h, the expression of A20 protein was increased at 2 h and 4 h, except that the expression of A20 protein in?creased after 6 h (P<0.05). Meanwhile NF-κB expression increased and IκB expression decreased in different time points (P<0.05). In addition, the expressions of A20 and IκB were decreased concentration-dependently (P<0.05). The expres?sion levels of NF-κB and MCP-1 were increased concentration-dependently (P<0.05). Conclusion A20 may involve in the development of diabetic nephropathy by regulating the NF-κB pathway.
6.High-dose methylprednisolone sodium succinate therapy for acute spinal cord injury in adults:a Meta-analysis
Xiangdong LIU ; Lei WANG ; Zhe DONG ; Yanhui MA ; Fang HU ; Gaiping MA ; Fengfeng WANG ; Shengjun YANG
Chinese Journal of Tissue Engineering Research 2015;(42):6875-6881
BACKGROUND:Some control studies attempt to answer the advantages and disadvantages of high-dose methylprednisolone sodium succinate therapy for acute spinal cord injury in adults, but have arrived at different conclusions. OBJECTIVE:To explore the therapeutic efficacy of high-dose methylprednisolone sodium succinate therapy on acute spinal cord injury in adults by Meta analysis. METHODS:PubMed, Embase, Cochranel Library, CBMdisc, VIP and WanFang Databases were searched by computer, and relevant Chinese and English orthopedic journals were retrieved by hand. Controled trials related to high-dose methylprednisolone sodium succinate therapy of acute spinal cord injury in adults were included. The methodology quality of included trials was criticaly assessed. RevMan 5.0 software was used for data analysis. RESULTS AND CONCLUSION:Nine clinical controled trials were included. Meta-analysis results showed that compared with the conventional therapy, the neurological recovery rate after 24 hours of administration, pneumonia incidence and gastrointestinal reactions increased significantly after high-dose methylprednisolone sodium succinate therapy. However, there were no statistical differences in the rate of urinary tract infection, nonunion rate and stress ulcer incidence between these two therapies. These findings indicate that the high-dose methylprednisolone sodium succinate therapy on acute spinal cord injury in adults has better outcomes in neurological function recovery, but can lead to higher incidence of lung infection and gastrointestinal reactions. Therefore, lung infection and gastrointestinal reactions should be avoided as much as possible during the course of treatment.
7.Meta analysis of clinical outcome of intramedullary nails versus locking plates for two-part proximal humerus fracture
Lei WANG ; Fengfeng WANG ; Yanhui MA ; Jie ZHANG ; Fang HU ; Gaiping MA ; Meimei LIU ; Zhangwen MA
Chinese Journal of Tissue Engineering Research 2017;21(3):478-484
BACKGROUND:Intramedul ary nails and locking plates are widely used for two-part proximal humerus fracture. Which is better for two types of implants in patients remains controversial. OBJECTIVE:To determine the clinical outcome of intramedul ary nails versus locking plates for two-part proximal humerus fracture according to Cochrane Meta analysis. METHODS:We searched PubMed, SCI, Embase, the Cochranel Library and CBMdisc, VIP information, Wanfang Database, and CNKI for randomized control ed trials and quasi-randomized control ed trials on intramedul ary nails and locking plates for two-part proximal humerus fracture. RevMan 5.2 software was used to analyze operation time, intraoperative blood loss, fracture healing time, postoperative complications (heterotopic ossification, pain, screw penetration, necrosis of humeral head), and Constant Score. RESULTS AND CONCLUSION:Six articles of clinical control ed trials were included with 259 patients. 131 patients received intramedul ary nails, and 128 patients received locking plates. Meta-analysis displayed that no significant difference in fracture healing time, heterotopic ossification, pain, necrosis of humeral head and Constant Score was detected between intramedul ary nails and locking plates for two-part proximal humerus fracture. Operation time, intraoperative blood loss, and screw penetration were significantly less in the fixation with intramedul ary nails than that in locking plates (P<0.05). These findings suggested that compared with locking plates, intramedul ary nails method for two-part proximal humerus fractures could reduce screw penetration.
8.Recombination of human HER2 gene encoding the extracellular domain and purification of fusion protein.
Xiaorong LIU ; Zhimin HE ; Fang YANG ; Yanhui YU ; Hui LV ; Zhuchu CHEN
Journal of Biomedical Engineering 2006;23(1):136-141
To purify the extracellular domain of HER2 in vitro and improve its prokaryotic expression abundance, the cDNA fragment encoding extracellular domain of HER2 was obtained by PCR and cloned into the expression vector pGEX-6P-1. After transforming it into Escherichia coli BL21, we instituted an investigation of different inducing conditions to try out the optimal condition for expressing soluble fusion protein. As for insoluble inclusion bodies, they were dissolved in 8 M Urea and refolded in refolding buffer. The soluble protein and the refolded protein were purified with Glutathione Sepharose 4B, respectively. The results showed that both the soluble and insoluble protein existed in Escherichia coli, but the majority was insoluble. It is beneficial to the expression of soluble fusion protein by induction at lower temperature (30 degrees C) and higher optical density (A600= 1.8) with the use of certain additive in medium. By purification of the supernatant of the lysate and refolded protein, the yield of the fusion protein was about 1.23 mg per liter culture. As a result, we have obtained the maximum soluble extracellular domain of HER2 protein, and thus have laid a foundation for further work on functional study and antibody preparation for HER2.
Cloning, Molecular
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Escherichia coli
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genetics
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Genes, erbB-2
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genetics
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Humans
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Prokaryotic Cells
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metabolism
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Protein Folding
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Receptor, ErbB-2
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biosynthesis
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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isolation & purification
9.Investigation on improving standard of polypropylene infusion bottles
Xiang LI ; Jinfeng ZHENG ; Fang LIN ; Yanhui XU ; Xia ZHAO
Drug Standards of China 2024;25(2):176-180
Objective:To explore the use of differential scanning calorimetry(DSC)method to identify polypropyl-ene infusion bottles material.Methods:The thermal analysis curve under nitrogen was determined at temperatures between 40 ℃ and 200 ℃ with a heating rate of 20 ℃·min-1,then cooled to 40 ℃ at a rate of 20 ℃·min-1.The sample was heated from 40 ℃ to 200 ℃ at a heating rate of 10 ℃·min-1,held the specimen at 200 ℃ for 10 min,then cooled to 40 ℃ at a rate of 10 ℃·min-1.Results:The melting peak temperature(Tm)of polypropyl-ene infusion bottles was between 145-150 ℃.Conclusion:The proposed method is accurate,sensitive,and sim-ple,and can be used for determination of polypropylene infusion bottles material,which can also provide reference for the standard improvement of polypropylene infusion bottles.
10.Investigation on dissolved substances of polypropylene port for plastic infusion containers
Fang LIN ; Yanhui XU ; Xiang LI ; Jinfeng ZHENG ; Xia ZHAO
Drug Standards of China 2024;25(4):416-422
Objective:To establish a preparation method for dissolved matter test solution of polypropylene port for plastic infusion containers,and to provide a reference for a standard system for pharmaceutical packaging materials of this product.Methods:Determination for results of dissolved matter test solution by specific surface area method and mass method were compared and analyzed in pair.Results:There were differences in sample amount and con-tact area between the two methods,but the results of dissolved matter test solution were generally consistent.Paired sample Wilcoxon signed rank test and paired sample T-test were used in pH,readily oxidizable substances,and non-volatile substance which indicators are quantifiable.Results show that there were no significant differences between the test solution,with Cohen's d values of 0.153,0.218,and 0.296,respectively,which were small in magnitude.Conclusion:In the case of insignificant differences between the two sampling methods,for polypropyl-ene port for plastic infusion containers and other irregular products,it is recommended that the test solution is prepared by leaching by mass,that is,a leaching ratio of 0.2 g·mL-1 by mass or volume,with more reproducible results.This study provides a reference for the establishment of standard for polypropylene port for plastic infusion containers.