Objective To evaluate the association between cytomegalovirus (HCMV) infection and atherosclerosis (AS) and explore the possible role of endothelin (ET) and serum tumor necrosis factor(TNF ?) in the pathogenesis of AS in diabetic patients and the relationship between HCMV infection and concentrations of ET and TNF ?. Methods Blood samples were collected from 21 patients with AS of type 2 DM, 47 patients with non AS of type 2 DM and 20 controls. Special antibodies to HCMV (IgM,IgG) were measured by enzyme linked immunosorbent assay (ELISA) . The concentrations of serum TNF ? and plasma ET were determined by RIA. Results (1) Type 2 diabetic patients, especially those with established AS, had a higher prevalence of active infection of HCMV. (2) The levels of plasma ET and serum TNF ? in type 2 diabetic patients were higher than those in normal people. In the pathogenesis of AS in diabetic patients, the ET played more important role than the TNF ?.(3) There was a significantly positive correlation between anti IgM to HCMV and the level of plasma ET in type 2 DM patients. It suggested that the injury of endothelium cell was related to the active infection of HCMV. Conclusion The study suggests that HCMV infection may be important in the pathogenesis of AS in type 2 diabetic patients and ET possibly plays an important role on the pathological course of AS.

Objective To improve the quality standard of Qixue Shuangbu Oral Liquid;To provide guarantee for the quality of oral liquids.Methods HPLC was used to simultaneously determine the contents of paeoniflorin and icariin in the oral liquid. The determination was performed on a Agiltnt TC-C18 (2) column (250 mm × 4.6 mm, 5μm) with the mobile phase of acetonitrile and 0.1% H3PO4 gradient mode. The flow rate was 1.0 mL/min;the column temperature was set at 25℃; the detection wavelength was 232 nm.Results The linear ranges of paeoniflorin and icariin were obtained between 0.542-8.677μg (r=0.999 9) and 0.185-2.963μg (r=0.999 9), with the average recoveries (n=6) of 99.29% (RSD=1.06%) and 98.91% (RSD=1.97%), respectively.Conclusion This method is accurate and feasible, which can be used conveniently in quality control ofQixue Shuangbu Oral Liquid.

Objective To investigate the efficacy and safety of citrate anticoagulation for continuous renal placement therapy (CRRT) in patients at high risk of bleeding. Methods Forty-seven patients who needed to CRRT and were admitted into the department of critical care medicine from January 2015 to October 2016 were enrolled in this study. According to the patient′s actual condition, they were divided into citrate groups (24 cases) and saline group (23 cases). Patients in saline group were given saline wash. The efficacy and safety were compared between the two groups. The filter lifetime, after treatment the activated partial thromboplastin time (APTT), hemoglobin (Hb), blood gas indexes were compared between the two groups. Results The citrate group used 76 filters while the 0.9% sodium chloride group used 87 filters. The average filter lifetime in citrate group was (22.4 ± 12.7) h which was longer than (8.6±3.3) h in 0.9%sodium chloride group (t=9.79, P<0.01). The incidence of coagulation in extracorporeal circulation was 3.9%(3/76) which was lower than 16.1%(14/87) in 0.9%sodium chloride group(χ2=5.20, P<0.05). Conclusions Regional citrate anticoagulation is a safe and effective option for CRRT in patients at high risk of bleeding.

BACKGROUND:In recent years, chitosan-based thermosensitive hydrogel, as scaffold materials, have received more and more attentions in the field of tissue repair because of good biocompatibility, biodegradability and drug-sustained release.
OBJECTIVE:To explore the directed differentiation and growth of rat bone marrow mesenchymal stem cells on the quaternary chitosan thermosensitive hydrogel scaffold and to look for more ideal tissue engineering materials for the treatment of nervous system damage.
METHODS:The thermosensitive hygrogel scaffold was prepared using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) andβ-glycerophosphate (β-GP). The spatial structure of scaffold was observed by scanning electronic microscope. Effect of leaching liquor from the HACC/β-GP scaffold on the viability of bone marrow mesenchymal stem cells was detected by (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The albumin from bovine serum was combined with the scaffold, and the slow-release effect of the scaffold was detected by ultraviolet absorption spectrometry. Bone marrow mesenchymal stem cells were incubated onto the compound scaffold at 3 passages. The adhesion, growth and differentiation of bone marrow mesenchymal stem cells on the compound scaffold were observed by the scanning electron microscope. Neuron-specific enolase was detected by immunofluorescence.
RESULTS AND CONCLUSION:The porosity and thermal sensitivity of HACC/β-GP scaffold and slow-release effect of glial cellline-derived neurotrophic factor were apparent. The results of MTT showed that the compound scaffold cannot take apparent negative effects to the proliferation of bone marrow mesenchymal stem cells. After inoculation, bone marrow mesenchymal stem cells permeated the porous structure of the scaffold and adhered to the scaffold. Under the role of glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells showed neuron-like cellmorphology and cells co-cultured with the compound scaffold expressed the marker of neurons, neuron-specific enolase. Under the role of slow-release glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells can grow wel in vitro and differentiate into neuron-like cells on the HACC/β-GP scaffold.

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

Objective:To establish the quality control of Zhiqikang capsules. Methods:TLC was used to identify Gastrodia tuder halimasch, rhubarb and Astragalus mongholicus in the preparations. A spectrophotometry method with 3, 5-dinitrosalicylic acid (DNS) was used to measure the polysaccharide content in Zhiqikang capsules. A spectrophotometry method with Forint phenol method ( Low-ry) was used to measure the peptide content in the capsules. Results:The linear range of polysaccharide was obtained between 6. 412 and 32. 060μg·ml-1(r=0. 999 5), the average recovery was 95. 86% and RSD was 0. 86%. The linear range of peptide was ob-tained between 0.059 7 and 0.298 4 mg·ml-1(r=0.999 0), the average recovery was 100.3% and RSD was 1.88%(n=6). Conclusion:The assay method is simple and accurate in the quality control of the preparations.

BACKGROUND:Scaffolds made of chitosan and its derivatives play an important role in cellmigration and axonal regeneration. Chitosan and its derivatives have good histocompatibility, which is easy to make stem cells grow on the surface, thereby having a more broad application prospect in the nerve tissue engineering.
OBJECTIVE:To fabricate a thermosensitive hydrogel scaffold using chitosan/hydroxypropyltrimethyl ammonium chloride chitosan/sodium glycerophosphate (CS/HACC/GP), which is suitable for cellgrowth, and then, to observe the growth and proliferation of bone marrow mesenchymal stem cells on the scaffold.
METHODS:Chitosan was modified using quaternary ammonium salt and confirmed by Fourier transform infrared spectroscopy. The chitosan and quaternary ammonium salt of chitosan was mixed at a ratio of 8:1 to successful y prepare stable CS/HACC/GP thermosensitive hydrogel scaffold. Then, the gel ing was observed, and biosafety test was conducted.
RESULTS AND CONCLUSION:Fourier transform infrared spectroscopy showed the characteristic peak of quaternary ammonium groups. Cytotoxicity test showed that rat bone marrow mesenchymal stem cells cultured in hydrogel extracts had no toxicity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test showed that hydrogel extracts exerted no significant effect on the increase in body weight, and the biological safety of the scaffold was good. Under the scanning electron microscopy, bone marrow-derived mesenchymal stem cells grew and proliferated normal y in the scaffold. The results confirmed that the CS/HACC/GP thermosensitive hydrogel scaffold was successful y prepared in the experiment, which is suitable for the growth and proliferation of bone marrow mesenchymal stem cells.

Objective To investigate the characteristics of ocular surface inflammation index after pterygium excision by using Oculus Keratograph(R) 5M and Visual analogue scale and evaluated the effectiveness of antiinflammatory treatment.Methods A prospective case control study was performed.Eighteen patients (6 males and 12 females) who suffered from primary pterygium were recruited in Zhongshan Ophthalmic Center from June to September 2016.All patients were treated with monocular pterygium excision combined with amniotic membrane transplantation.Anti-inflammatory treatment was given after surgery,and the ocular inflammation index was evaluated at preoperative and 1st,3rd,7th,10th,30th and 60th day postoperative.The temporal conjunctival hyperemia index (TCHI) was assessed by Oculus Keratograph(R) 5M with a red eye index analysis software.Ocular symptom scores (OSS) and visual analogue scale (VAS) were used to analyze the subjective symptoms of the patients.Fluorescein staining was used to detect the epithelization of corneal and scleral wound.The best corrected visual acuity (BCVA),intraocular pressure and complications were evaluated in this study.This study was approved by the Medical Ethics Committee of Zhongshan Ophthalmic Center of Sun Yat-sen University (2016KYPJ024).All patients signed informed consent for clinical research.Results No drug-related ocular and systemic adverse events were found during the follow-up.Corneal epithelial defect was recovered on 10th day,and conjunctival epithelization was observed in sclera exposed area on 30th day.The BCVA on the 60th day was 0.12±0.17,which was significantly lower than 0.34±0.36 preoperatively (t =3.401,P =0.003).Compared with those before surgery,OSS and VAS were significantly increased on 1 st day (OSS:Z =-4.255,P =0.000;VAS:Z =-5.256,P =0.000).The OSS on 7th day was not significantly different from that before surgery (Z=-0.958,P=0.372).VAS decreased to baseline on 30th day.The OSS on 60th day after surgery was significantly lower than that before surgery (Z =-2.397,P =0.037).TCHI was higher than 1.2 preoperatively,and increased to the highest on 1 st day after surgery,with significant difference between them (t=-6.620,P=0.000).The TCHI decreased to baseline on 7th day,no significant difference were obtained when compared with preoperative TCHI (t =-1.050,P =0.310),and TCHI on 60th day after surgery was lower than that before surgery,with significant difference between them (t =2.758,P =0.020).Conclusions The subjective symptoms combined with conjunctival hyperemia can be more accurate assessment of ocular surface inflammation in the perioperative period of pterygium surgery,which can be used as an evaluation index to assess the effectiveness of anti-inflammatory treatment.

Objective:To compare the clinical value of stent assisted intestinal bypass and temporary loop ileostomy in laparoscopic low anterior resection of rectal cancer.Method:In this retrospective analysis, 57 patients undergoing laparoscopic low anterior resection for rectal cancer in the First Affiliated Hospital of Ningbo University from Jan 2020 to Jan 2022 were divided into intestinal bypass group (36 cases) and loop ileostomy group (21 cases).Result:There were no significant differences in postoperative GI function recovery and postoperative complication rate between the two groups (all P>0.05). The levels of albumin, prealbumin and hemoglobin in the intestinal bypass group were better than those in the ileostomy group when evaluated on 3rd months after operation [(40.5±2.3) g/L vs. (38.1±2.6)g/L、(26.4±2.7)mg/dl vs. (24.5±2.0)mg/dl、(137.6±5.9) g/L vs. (134.0±7.0) g/L, t=3.605、2.743、2.085, all P<0.05]. Hospital expenses of the intestinal bypass group was lower [(571 000±7 500) yuan vs. (69 300±9 100) yuan, t=-5.477, P<0.05]. Conclusion:Compared with traditional ileostomy, the stent assisted intestinal bypass reduces trauma with lower expenses and improves patients' status after laparoscopic low anterior resection for rectal cancer.

The axon initial segment (AIS) is a specialized structure that controls neuronal excitability via action potential (AP) generation. Currently, AIS plasticity with regard to changes in length and location in response to neural activity has been extensively investigated, but how AIS diameter is regulated remains elusive. Here we report that COUP-TFI (chicken ovalbumin upstream promotor-transcription factor 1) is an essential regulator of AIS diameter in both developing and adult mouse neocortex. Either embryonic or adult ablation of COUP-TFI results in reduced AIS diameter and impaired AP generation. Although COUP-TFI ablations in sparse single neurons and in populations of neurons have similar impacts on AIS diameter and AP generation, they strengthen and weaken, respectively, the receiving spontaneous network in mutant neurons. In contrast, overexpression of COUP-TFI in sparse single neurons increases the AIS diameter and facilitates AP generation, but decreases the receiving spontaneous network. Our findings demonstrate that COUP-TFI is indispensable for both the expansion and maintenance of AIS diameter and that AIS diameter fine-tunes action potential generation and synaptic inputs in mammalian cortical neurons.
Action Potentials ; Animals ; Axon Initial Segment ; COUP Transcription Factor I ; DNA-Binding Proteins/physiology* ; Mammals ; Mice ; Transcription Factors