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.

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.

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.

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.

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.

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.

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.

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 ; Escherichia coli ; genetics ; Genes, erbB-2 ; genetics ; Humans ; Prokaryotic Cells ; metabolism ; Protein Folding ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Objective:The aims of the study were to investigate the relationship among atherogenic index of plasma (AIP) and inflammatory adipocytokines with the severity of coronary artery calcification (CAC) score in coronary artery disease (CAD). And then we analyzed the diagnostic value of the new markers on CAC.Methods:A total of 241 patients with CAD diagnosed by coronary CT angiography (CTA) and coronary angiography in Baoding First Central Hospital from June 2019 to June 2020 were retrospectively enrolled. According to the presence of calcification in coronary CTA, they were divided into CAC group ( n=63) and non-CAC group ( n=178). The clinical data of the patients were collected, and the levels of serum inflammatory factors were measured by enzyme-linked immunosorbent assay (ELISA). The correlation between CAC score and AIP and inflammatory cytokines was analyzed. The diagnostic value of AIP and inflammatory factors in the formation of CAC in patients with CAD. Results:The levels of AIP, serum osteoprotegerin (OPG) and oligomeric matrix protein (COMP) in CAC group were higher than those in non-CAC group, while the levels of serum fibroblast growth factor 21 (FGF21) were lower than those in non-CAC group, with statistically significant difference (all P<0.01). Correlation analysis showed that CAC score of CAD patients was positively correlated with AIP, OPG and COMP ( r=0.581, 0.451, 0.326, P<0.05), and negatively correlated with FGF21 ( r=-0.294, P<0.05). Receiver operating characteristic (ROC) curve analysis showed that AIP, OPG, COMP and FGF21 had diagnostic value for CAC in CAD patients (all P<0.05). AIP>0.387, OPG>5.150 ng/ml, FGF21>136.35 pg/ml, COMP>733.16 ng/ml were independent factors affecting the formation of CAC (all P<0.05). Conclusions:The increase of AIP and the change of inflammatory factors can be used as markers for the diagnosis of CAC formation in CAD patients.

OBJECTIVE: To analyze the phenotype and genetic variants of a pedigree affected with hereditary protein C (PC) deficiency. METHODS: The protein C activity (PC:A) of the proband and her family members (a four-generation pedigree including 11 individuals) were tested by chromogenic substrate method, and the protein C antigen (PC:Ag) was detected with an enzyme-linked immunosorbent assay(ELISA). The 9 exons and flanking sequences of the protein C (PROC) gene were amplified by PCR and directly sequenced. Suspected mutation was validated by clone sequencing and in other members of the family. MutationTaster and ClustalX-2.1-win was used to analyze the pathogenicity and conservation of the mutation site,respectively. Three-dimentional protein model and amino acids interaction were analyzed with Swiss-PdbViewer software. RESULTS: The PC: A and PC: Ag of the proband were decreased to 46% (reference range: 70%-130%) and 50% (referencerange:70%-140%), respectively. Her grandmother,aunt, cousin and younger brother also showed declined PC:A and PC:Ag by approximately 50%. Genetic analysis revealed that the above individuals have all carried a deletional mutation c.1212-1212delG (p.Met364TrpfsX15) in exon 9 of the PROC gene which can cause replacement of Methionine at position 364 by Tryptophan and alteration of 15 downstream amino acids, and produce a premature stop codon at position 378. The score of MutationTaster was 1.000, indicating that the variant is pathogenic. Conservative analysis showed that the 15 altered amino acids are located in a conserved region across nine homologous species. Protein model analysis showed that the mutation has disrupted a catalytic domain of protein C thereby affected its function. CONCLUSION The heterozygous c.1212-1212delG deletional mutation in exon 9 of the PROC gene, which was unreported previously,probably accounts for the decrease of PC:A and PC:Ag in this pedigree.