BACKGROUND:Traditional Chinese medicine has rich experience and unique advantages in the empirical treatment of phlegm-heat and Fu-organs excess syndrome of ischemic stroke.In order to further explore the therapeutic targets and mechanisms of traditional Chinese medicine for this disease,it is crucial to establish a stable and reliable animal model of phlegm-heat and Fu-organs excess syndrome combined with empirical symptoms of ischemic stroke. OBJECTIVE:To explore the establishment method and evaluation system of the rat model of ischemic stroke with phlegm-heat and Fu-organ excess syndrome. METHODS:Sixty male Sprague-Dawley rats were randomly divided into four groups:blank control group(n=12),ischemic stroke group(n=18),disease+syndrome group(n=18),phlegm-heat and Fu-organ excess syndrome group(n=12),all of which were given high-fat diet for 25 days.On the 26th day,the rats in the blank control group and ischemic stroke group were intragastrically given normal saline and high fat diet,while those in the other two groups were intragastrically given autologous feces suspension and high fat diet for 3 continuous days.After gavage,ischemic stroke models were established using the suture method in the ischemic stroke group and disease+syndrome group.The changes in diet,water intake,body mass,body temperature,fecal traits,nasal secretions,sputum in the throat,and tongue image were recorded.Neurological deficits,tongue image,blood lipid levels,morphological changes of brain tissue and carotid artery,and the serum levels of motilin and somatostatin were detected. RESULTS AND CONCLUSION:Compared with the control group,the rats in the disease+syndrome group had shortness of breath,listlessness,irritability,bradykinesia,a large number of secretions around the nose,audible and heavy sputum in the throat,decreased diet and water intake,increased body mass,body temperature,and slingual vein score,decreased fecal pellet count,Bristol score and fecal moisture content,increased serum total cholesterol,triglyceride,low-density lipoprotein and somatostatin levels,decreased motilin level,increased neurological deficit score,significant pathological changes of the carotid artery,and significant morphological changes of the brain tissue.The ischemic stroke group only showed pathological changes of ischemic brain tissue,without the characteristics of phlegm-heat and Fu-organ excess syndrome.The phlegm-heat and Fu-organ excess syndrome group could present with the typical characteristics of traditional Chinese medicine syndromes,without the pathological changes of brain tissue with ischemic stroke.To conclude,the compound modeling method of high-fat induction combined with suture method and autologous feces gavage can establish an animal model of ischemic stroke with phlegm-heat and Fu-organ excess syndrome.

The formation of most proteins consists of two steps: the synthesis of precursor proteins and the synthesis of functional proteins. In these processes, propeptides play important roles in assisting protein folding or inhibiting its activity. As an important polypeptide chain coded by a gene sequence in lipase gene, propeptide usually functions as an intramolecular chaperone, assisting enzyme molecule folding. Meanwhile, some specific sites on propeptide such as glycosylated sites, have important effect on the activity, stability in extreme environment, methanol resistance and the substrate specificity of the lipase. Studying the mechanism of propeptide-mediated protein folding, as well as the influence of propeptide on lipases, will allow to regulate lipase by alternating the propeptide folding behavior and in turn pave new ways for protein engineering research.
Lipase/metabolism* ; Molecular Chaperones/metabolism* ; Protein Folding ; Protein Precursors ; Substrate Specificity

Objective:To investigate the relationship between peritoneal thickness and baseline solute transport function in peritoneal dialysis (PD) patients, and analyze the factors affecting the function of peritoneal transport.Methods:Non-diabetic end-stage renal disease (ESRD) patients admitted to the Second Hospital of Longyan City from January 2017 to June 2019 were enrolled in this study. The thickness of the peritoneal membrane was measured by color ultrasound instrument before the peritoneal catheterization. Standard peritoneal equilibration test (PET) was performed after one month of peritoneal dialysis. The ratio of corrected creatine in 4 h dialysate to 2 h serum creatine (D/Pcr) was used as a solute baseline transport index, and according to the D/Pcr evaluation results, the patients were divided into high/high average transfer (H) group (D/Pcr≥0.65) and low/low average transfer (L) group (D/Pcr<0.65). The clinical data, peritoneal thickness and peritoneal dialysis related indicators between the two groups of patients were compared. Binary logistic regression was used to analyze the factors affecting the function of peritoneal transport.Results:The amount of peritoneal ultrafiltration in H group was significantly lower than that in L group, intraperitoneal creatinine clearance (Ccr) and peritoneal thickness were significantly higher than those in L group (both P<0.05). Pearson and Spearman correlation results showed that the thickness of peritoneal membrane positively correlated with D/Pcr ( r=0.673, P<0.05), peritoneal Ccr ( r=0.261, P<0.05), and negatively correlated with ultrafiltration of peritoneal dialysis ( r=-0.365, P<0.05). Partial correlation analysis showed that the peritoneal thickness was positively correlated with the solute transport index D/Pcr ( r=0.539, P<0.05) and the peritoneal Ccr ( r=0.338, P<0.05). Binary logistic regression results showed that peritoneal thickening was a risk factor affecting peritoneal transport function ( OR=1.175, 95% CI 1.009-1.369, P<0.05). Conclusions:There is a positive correlation between the peritoneal membrane thickness and the baseline solute transport index in patients with non-diabetic peritoneal dialysis. Peritoneal thickening is a risk factor affecting peritoneal transport function.

Objective To analyze the clinical features of patients with acute cerebral infarction combined with periodic limb movements during sleep (PLMS). Methods A total of 170 continuous patients with acute cerebral infarction in the Department of Neurology of the Second Affiliated Hospital of Soochow University from February 2016 to June 2018 was enrolled prospectively, including 102 males (60. 0%) and 68 females(40. 0%). Those patients were divided into non-PLMS group (period limb movement index[PLMI] <5 times/hour) and PLMS group (PLMI ≥5 times/hour) according to the PLMI. Physical activity recorder monitoring and head magnetic resonance imaging were performed within 2 weeks of onset. The baseline National Institutes of Health Stroke Scale(NIHSS) scores, previous medical history (including hypertension, diabetes, atrial fibrillation, coronary heart disease, hyperlipidemia), smoking history, stroke classification of the Oxfordshire Community Stroke Project(OCSP), biochemistry indicators (glycated hemoglobin, creatinine, homocysteine, total cholesterol, low-density lipoprotein), and sleep parameters were recorded and compared between both groups. The modified Rankin Scale (mRS) score was used to evaluate the recovery of neurological function at 3, 6, and 12 months, and the Barthel index was used to assess the self-care ability of daily life at 12 months. PLMI ≥5 times/h was defined as PLMS. Results (1) The proportion of patients with acute cerebral infarction combined with PLMS was 61.2% (104/170). The proportion of previous stroke history in the non-PLMS group was lower than that in the PLMS group, which was statistically significant (24.2% [16/66] vs. 39.4% [41/104], χ2=3.866, P = 0.049). There were insignificant in age, gender, smoking history, hypertension, diabetes, atrial fibrillation and NIHSS scores between two groups (all P > 0. 05). (2) In the non-PLMS group, the proportion of partial anterior circulation infarction was the highest (34. 8%, 23/66), and the proportion of complete anterior circulation infarction was the lowest (10. 6%, 7/66). In the PLMS group, the proportion of lacunar infarction was the highest (36.5 %, 38/104), and the proportion of complete anterior circulation infarction was the lowest (8.7%, 9/104). There was statistically significant in stroke classification of OCSP between the two groups (χ2=12.528, P=0.006), but insignificant in fasting blood glucose, glycated hemoglobin, creatinine, homocysteine, total cholesterol and low-density lipoprotein levels between the two groups (both P > 0. 05). (3) The proportion of awakening, awakening time, and the number of awakenings in the non-PLMS group were all lower than those in the PLMS group, which were statistically significant (6.0[3.0, 8.0] vs. 12.0[7. 0, 19.0], 3.0[1.5, 4.2] min vs. 4.4[3.0, 6.0] min and 18.5[7.0, 33.8] times vs.50.0[28.0, 84.0] times, the Z values were -6.046, -3.922 and -6.8789, all P < 0.01). Sleep efficiency and total sleep time were higher in the non-PLMS group than those in PLMS group, which were statistically significant (96. 6% [93. 6%, 98. 7%] vs. 91. 0% [84. 6%, 93. 9%], 497.0[470.0, 529.8]minus. 489.0[447.0, 507.0]min, and the Z values were -6.586 and -2.503 respectively, all P < 0. 01). (4) There was insignificant in the rate of good prognosis at 3, 6, and 12 months, and Barthel index at 12 months after onset between both groups (all P > 0. 05). Conclusions Patients with acute cerebral infarction have a high proportion of PLMS in the acute phase. There is a high proportion of lacunar infarction and sleep fragmentation in patients with acute cerebral infarction combined with PLMS.

The standardized residency training has become a necessary way to improve the overall professional quality of doctors. At present, a single teaching model can't meet the needs of the standardized residency training. This study explores the application of problem-based learning (PBL) combined with multidisciplinary treatment (MDT) model in the general surgery teaching of standardized residency training, so that the residents can use their theoretical knowledge to think deeply about the problems in the cases. By consulting the literature review, the cases are analyzed from multiple levels and angles, such as pathogenic causes, pathogenesis and clinical manifestations, then the clinic diagnosis and therapeutic schedule can be obtained. This kind of teaching model not only stimulates the group's more interest in learning, and improves the ability of autonomous learning, independent analysis, problem solving and language expression, but also significantly improves teaching satisfaction and has obvious teaching advantages.

OBJECTIVE: To study the effect of aquaporin 4(AQP4) in regulating the permeability of blood-brain barrier(BBB) induced by subacute 1,2-dichloroethane(1,2-DCE) inhalation. METHODS: Specific pathogen free healthy CD-1 male Aqp4 genetically engineered mice(Aqp4~(+/+)and Aqp4~(-/-)) were randomly divided into control and low-, medium-and high-dose groups. The mice were exposed to 1,2-DCE at the dosages of 0.00, 100.00, 350.00 and 700.00 mg/m~3 for 6 hours per day for consecutive 28 days by systemic dynamic inhalation. After the end of 1,2-DCE exposure, the BBB permeability was evaluated by Evans blue staining. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of genes related to BBB tight junction protein(Tjp)1, Tjp2, Tjp3, claudin(Cldn)3, Cldn5, Cldn11, occludin(Ocln), matrix metalloproteinase(Mmp)2, Mmp9 and Na-K-Cl cotransporter-1(Nkcc1). RESULTS: The BBB permeability in mice showed significant change with 1,2-DCE dose and Aqp4 genotype(P<0.01). The BBB permeability of Aqp4~(+/+) genotype mice was higher in low-, medium-and high-dose groups than that of control group(all P values were <0.05). The permeability of BBB was lower in Aqp4~(+/+) genotype mice in the control group than that of Aqp4~(-/-) genotype mice in the same group(P<0.05), but BBB permeability was higher in Aqp4~(+/+) genotype mice in the three dose groups than that of Aqp4~(-/-) genotype mice in the same group(all P values were <0.05). The Cldn3 and Olcn mRNA relative expression in the brain cortex had statistical difference in mice with different genotype(all P values were <0.01). The mRNA relative expressions of Cldn3 and Olcn in the brain cortex were higher in Aqp4~(-/-) genotype mice than that of Aqp4~(+/+) genotype mice(all P values were <0.01). The relative mRNA expression levels of Tjp1, Tjp2, Tjp3, Cldn5, Cldn11, Mmp2, Mmp9 and Nkcc1 in the cerebral cortex of mice were not statistically significant in aspect of 1,2-DCE exposure dose and genotype(all P values were >0.05). CONCLUSION: Exposure to 1,2-DCE can increase BBB permeability in mice, and the mechanism may be associated with 1,2-DCE-induced down-regulation of Aqp4 and up-regulation of mRNA expression of the cerebral cortex TJP-related molecules Cldn3 and Ocln.

Objective To explore the effect of acupuncture plus neuromuscular electrical stimulation and cold stimulation on children with swallowing disorders caused by viral encephalitis.Methods A total of 322 chil dren with dysphagia subsequent to viral encephalitis were randomly divided into an acupuncture + cold stimulation group (group A,n=107),an electrical stimulation + cold stimulation group (group E,n=107) and an acupuncture + electric stimulation + cold stimulation group (group C,n=108) using a random number table.The treatments were applied in two courses each of 14 days with an interval of two days between the courses.Before and after the treatment,the total effectiveness rates and the incidence of aspiration pneumonia and malnutrition were ana lyzed.The children's swallowing function was evaluated using a standardized swallowing assessment (SSA),while the severity of dysphagia was assessed using dysphagia outcome and severity scale (DOSS) based on videofluorescopic imaging.Results There were no significant differences in any of the outcome measures before the treatment.Afterward the total effectiveness rate in group A was 76％,significantly higher than in group E (62％) but significantly lower than in group C (91％).There was significant improvement in the average SSA and DOSS scores of all 3 groups.Compared with group A,the average SSA score of group E was higher while the average DOSS score was lower.For group C the relationship was the opposite,with the average SSA score significantly higher and the average DOSS score significantly lower.After the treatment there were no significant differences between groups A and E in the incidence of aspiration pneumonia or malnutrition,but the incidence of both in group C was significantly lower than in groups A and E.Conclusion Acupuncture administered together with electrical and cold stimulation can significantly improve dysphagia and lower the incidence of aspiration pneumonia and malnutrition for children with viral encephalitis.

Objective: To investigate the effect of ubiquitous mitochondrial creatine kinase 1(CKMT1) on the sensitivity of human nasopharyngeal carcinoma cell line CNE-1 to DDP. Methods: CNE-1 cells were transiently transfected with CKMT1 overexpression (CKMT1) or empty vector (EV). The growth curve and DDP IC50 were developed by MTT assay, plate clone formation assay was performed by gradient concentration of DDP treatment, cell cycle and apoptosis were detected by flow cytometry, levels of apoptosis related protein Bax/Bcl-2/C-PARP and the transcription factor p-STAT3-Tyr705 were detected by Western Blot. Results: The transfection efficiencies of CKMT1 and EV were more than 90% with a higher proliferation rate in the CKMT1-transfected cells. However, the CKMT1-transfected cells had a DDP IC50 of 2.76 μmol/L, which was significantly lower than that of 4.60 μmol/L in the EV-transfected cells (P<0.01). With the treatment of certain concentration of DDP, the CKMT1-transfected cells had a lower clone formation rate, the cell cycle arrested more obviously in G2/M phase, and the apoptosis rate was higher (P<0.01), with higher levels of Bax/C-PARP (P<0.05 or P<0.01), but lower levels of Bcl-2 (P<0.01) and p-STAT3-Tyr705 (P<0.01), compare with the EV-transfected cells. Conclusions CKMT1 may inhibit the activation of STAT3, increasing the sensitivity of CNE-1 to chemotherapeutic drug DDP.

Objective To investigate the distribution of common carbapenem resistance genes and virulence genes in and understand the molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains.Methods A total of 84 non-duplicate CRKP isolates were collected from Renji Hospital,Shanghai Jiao Tong University School of Medicine and Changzheng Hospital,the Second Military Medical University,in 2015.Kirby-Bauer disk diffusion method was used to test their susceptibility to 15 antimicrobial agents.The HM phenotype of K.pneumoniae was determined by string test.Carbapenem-resistant genes and virulence genes were detected by polymerase chain reaction (PCR).The molecular epidemiology of the 84 isolates were further analyzed by multi locus sequence typing (MLST).The population structure of CRKPs was evaluated by eBURST with the results of MLST.Results Antimicrobial susceptibility test showed that 84 isolates were highly resistant to most antimicrobial agents such as carbapenems,penicillins,cephalosporins and aztreonam.More than 90％ of the strains were resistant to most of the antibiotics tested except ciprofloxacin (77.4％,65/84) and amikacin (82.1％,69/84).Two strains showed HM phenotype.PCR results showed that 90.5％ (76/84) of the strains were positive for blaKPC-2,1.2％ (1/84) positive for blaNDM and blaIMP each,but either blaOXA or blaVIM was not identified.The overall prevalence of virulence genes was low except for mrkD (97.6％,82/84),ybtS (92.9％,78/84) and entB (100％,84/84).Eight sequence types (STs) were obtained.The dominant clone was ST11 (84.5％,71/84),and the two strains of HM phenotype were ST11.eBURST analysis identified 2 ST groups among the 84 CRKPs.Each ST group includes 2 ST types (ST11 and ST1869,ST15 and ST709),respectively.The other four ST types were single ST type.In this study,71 strains of ST11,4 ST15 and 4 ST323 belonged to CC258,CC15 and CC163 clones,respectively.Conclusions CRKP is highly resistant to the commonly used antibiotics.The multidrug resistance or pandrug-resistance of K.pneumoniae is mainly associated with the expression of blaKPC-2 gene.Three virulence genes mrkD,ybtS and entB are highly prevalent in the CRKP isolates.The dominant clone of KPC-producing K.pneumoniae is ST11 in both hospitals.

OBJECTIVE: To explore the molecular mechanism underlying 1,2-dichloroethane(1,2-DCE) induced apoptosis by screening differentially expressed proteins in human astrocytes( HAs). METHODS: HAs were cultured in complete medium with 1,2-DCE at various concentrations of 0-80 or 0-40 mmol/L. After 24 hours,apoptosis of HAs was evaluated using flow cytometry and staining with annexin Ⅴ-fluoresce in isothiocyanate and propidium iodide. An AAH-APO-1-2 protein chip was used to screen differentially expressed proteins and quantitative real-time polymease chain reaction(qRT-PCR) was used to verify related differentially expressed genes(DEGs). RESULTS: At 1,2-DCE concentrations of0-80 mmol/L,the total apoptosis rate of HAs increased with 1,2-DCE concentrations in a dose-dependent manner( P <0. 01). Seven different kinds of proteins were screened out by apoptotic protein chip. Among them,the expression of insulin-like growth factor-binding protein( IGFBP)-1,IGFBP-4 and cytochrome C( Cyto C) were up-regulated,while the expression of P27,cysteine aspartic acid specific protease-3( Caspase-3),B-cell lymphoma-2 interacting mediator of cell death( BIM) and BH3 interacting domain death agonist( BID) were down-regulated compared with the control group. The result of DEGs verified by qRT-PCR showed that the expression of mRNA of IGFBP-1,IGFBP-4 and Cyto C at 1,2-DCE concentrations of 40 mmol/L was up-regulated. This result was in consistent with the trend of target expression in the protein chip. The mRNA expression of Caspase-3,BIM and BID was also up-regulated. CONCLUSION: 1,2-DCE induces apoptosis of HAs through mitochondrial pathway.