BACKGROUND: Dystrophin gene is X-linkage recessive heredity nerve-muscle system disease. Dystrophin gene deletions cluster in two hotspot regions, comprising exons 2-20 and 44-53. The majority of deletions can be detected by examining only a subset of exons. However, little is known regarding systematic detection of 18 common deletion exons of dystrophin gene.OBJECTIVE: To obtain and identify the cloning of 18 deletion-prone exons of dystrophin gene.METHODS: A total of 18 fragments of dystrophin gene were obtained through polymerase chain reaction (PCR) amplification with human genomic DNA as template and 18 pairs of primers respectively. The fragments were connected with pGEM-T Easy vector.The recombinants were transformed into E.coli JM109 competent cells, followed by planted on Luria-Bertani (LB)/ampicillin(Amp)/isopropylthio-β-D-galactoside(IPTG)/X-bromo-4-chloro-3-indolyl-β-D-galactoside (X-Gal) plates and cultured.Positive transformants were selected with blue/white color screening, and the recombinant plasmids DNA was extracted and digested with restriction enzyme Not I. DNA sequences of the fragments were analyzed. Nucleotide analyses were performed through the National Center for Biotechnology Information (NCBI) Basic Local Alighment Search Tool (BLAST) against GenBank.RESULTS AND CONCLUSION: Size of the18 fragments by PCR amplification was in accordance with anticipation. Size of the fragments of recombinant cloning by Not I digestion was in accordance with that of PCR and expectation. Sequence size of the 18cloned fragments was in accordance with expectation. The cloned fragments have high homology with dystrophin gene through NCBI BLAST against GenBank. These cloned fragments were the main deletion-prone exons of dystrophin gene.

OBJECTIVE:To investigate the permeability of lomefloxacin through blood-pancreatic barrier in rats.METHO-DS:Lomefloxacin(20mg/kg body weight) was injected through caudal vein.At the given time points,the samples were collected.The concentrations of lomefloxacin in the serum,pancreatic tissue and liver tissue were measured by HPLC.RESULTS:The concentration-time profiles of lomefloxacin could be described as two-compartment model in rats.The peak concentrations in serum,pancreatic tissue and liver tissue were 65.550?g/ml,48.801?g/g and 84.121?g/g at 5 min post-injection respectively.Then the concentrations decreased quickly in all of them.Concentrations in pancreatic tissue were higher than those in serum at 10 min and even at 480 min post-injection.The permeation ratio (PR) through blood-pancreatic barrier was 0.744 at 5 min and rose to 3.817 at 480min.CONCLUSION:After intravenous injection,lomefloxacin can permeate blood-pancreatic barrier satisfactory,therefore it is worthy of being recommended for prevention and treatment of pancreatic infections.

Objective:To observe immunological indexes,the quantity of cytokine expression and clinical curative effect of umbilical cord mesenchymal stem cells between before and after the treatment of systemic lupus erythematosus patients.Methods:Selected 10 cases of SLE,on the basis of glucocorticoid and immune inhibitor treatment,intravenous injection UC-MSC of cultivating proliferation within 6 generations.Before and after treatment of UC-MSC testing the relative quantity of cytokine of CTLA-4,IL-15,IL-2,CD86,IL-17c,Foxp3,TGF-β2 which were related of immunopathogenesis of SLE.Before and after treatment to determined SLE disease activity index (SLEDAI) score and detection of blood in the urine routine,liver and kidney function,24 hours urinary protein quantitative,immunoglobulin and complement levels.Results:After treatment the relative expression value of IL-15 and IL-2 was decreased,CTLA-4 was risen.There had no significant difference with the relative expression value of CD86,IL-17c,Foxp3,TGF-β2 in before and after treatment of UC-MSC.After treatment serum complement C3 and C4 level,serum albumin,were risen.24 hour proteinuria and SLEDAI were decreased.There was no serious adverse reaction occurred,no complications related to transplantation in 10 cases.Conclusion:UC-MSC can regulate the expression of cytokines of participate in the immune response in the patients with SLE.Treatment of SLE by UC-MSC can elevate serum albumin and C3 and C4 level,reduce the 24 hours urinary protein quantity,relife kidney damage,improve clinical symptoms;UC-MSC transplantation in patients with SLE have good security;UC-MSC transplantation may be a feasible method for the treatment of SLE.

Objective The aim of this study was to establish an acute gastric mucosal injury(AGMI)rat model induced by hydrochloric acid(HCl)at different concentration gradients and to investigate the effects of intravenous injection of evans blue(EB)at various concentrations and dosages on survival rate and superficial extravasation.Methods(1)Wistar rats were randomly assigned to five weight-based groups:150～180 g,180～200 g,200～250 g,300～400 g,400～500 g.Each group was further subdivided into eight subgroups based on HCl concentration,specifically:0.40,0.45,0.50,0.55,0.60,0.65,0.70 mol/L HCl,along with a control group treated with saline.This result ed in a total of 40 subgroups,with three rats per subgroup,summing up to 120 animals in total.The survival rate of rats 24 hours post-modeling was assessed,and the interaction between body weight and HCl concentration on rat survival was analyzed.Following the establishment of five graded HCl concentrations,gastric mucosal pathological changes were observed microscopically using hematoxylin-eosin staining(HE)staining.(2)From these result,the highest suitable concentration of HCl was selected to prepare the AGMI model.Subsequently,rats were randomly allocated into different groups based on the concentration and dose of EB,specifically:EB 1(0.5％,0.4 mL),EB 2(1％,0.1 mL/100 g),EB 3(1％,0.2 mL/100 g),EB 4(2％,0.1 mL/100 g),EB 5(2％,0.2 mL/100 g),and EB 6(5％,0.1 mL).Each group consisted of five rats,totaling 30 animals.The survival rate and extent of dermal exudation were evaluated 24 hours post-injection of EB.Results(1)Post-modeling symptoms in AGMI rats intensified with increasing concentrations of HCl,with the 24-hour survival rate in all weight groups being 0％for both 0.65 mol/L and 0.70 mol/L HCl.Kaplan-Meier survival curves indicated significant differences in survival rates among rats in different HCl concentration groups(P＜0.001).Furthermore,a significant interaction effect between HCl concentration and body weight on rat survival time was observed(P＜0.001).The five gradient HCl concentrations established were from 0.40、0.45、0.50、0.55、0.60 mol/L.Histological observations revealed that the degree of inflammatory cell infiltration in the gastric mucosa of AGMI rats escalated with increasing HCl concentrations.(2)AGMI rats prepared with 0.60 mol/L HCl and injected with 5％EB(0.1 mL)had a 24 h survival rate of only 40％.Kaplan-Meier survival curves showed no significant differences in survival rates among AGMI rats under different concentrations and dosages of EB(P＞0.05).Analysis of superficial extravasation revealed that skin and eye color were lighter in EB 1 group rats,with fewer extravasation points,while rats in EB 4 and EB 5 groups exhibited more apparent skin color changes and extravasation.One-way ANOVA further confirmed that the number of superficial EB extravasation points in the EB 3,EB 4,and EB 5 groups was significantly higher than that in EB 1 group(P＜0.05,P＜0.01).Conclusions HCl modeling successfully achieved a setting of multiple precise concentration gradients.Gastric lavage with HCl concentrations ranging from 0.40 mol/L to 0.60 mol/L in rats with a fasting body weight of 180～200 g was used to successfully prepare an AGMI model.Intravenous injection of 2％EB(0.2 mL/100 g)can facilitate the study and analysis of the distribution characteristics of superficial EB extravasation points in AGMI rats over time and as the condition progresses.

A 69-year old female patient was admitted because of 3 days of worsened chest pain.Coronary angiography showed60% stenosis of distal left main stem,chronic total occlusion of left anterior descending (LAD),70% stenosis at the ostium of a smallleft circumflex,70-90%stenosis at the paroxysmal and middle part of a dominant fight coronary artery (RCA),and a normal left internalmammary artery (LIMA) with normal origination and orientation.Percutaneous intervention was attempted but failed on the occludedlesion of LAD.The patient received minimally invasive direct coronary artery bypass (MIDCAB) with left LIMA isolation by Davincirobot.Eleven days later,the RCA lesion was treated by Sirolimus Rapamicin eluting stents implantation percutaneously.Then thepatient was discharged uneventfully after 3 days hospitalization.Our experience suggests that two stop shops of hybrid technique befeasible and safe in the treatment of elderly patient with multiple coronary diseases.

Hospice care is the product of the development of social civilization. Our country has gradually promoted the development of hospice care from the government, society, education and other levels, but has invested less in systematic education. This article reviews the development status of hospice care education at home and abroad in order to provide a reference for the development of related education in our country.

The existing doctor-patient communication pattern often falls prey to insufficient informed consent and even medical disputes. In the patient centered perspective, Zhejiang Provincial People′s Hospital explored a new communication mode centering on patients. Based on diagnosis-related groups catalogues and high-frequency surgeries catalogues of the departments, multimedia technology was called into play to produce dubbed PPTs and videos that were easy to understand, standardized and homogeneous, which were embedded into medical records system. Following observation of the PPT or video, patients could directly sign an informed consent on the computer. This practice not only deepens patient′s understanding and achieves homogeneous level of the communication, but also elevates doctor′s work efficiency, contributing to building a harmonious doctor-patient relationship.

Objective:To analyze the correlation between nutritional status and acute toxicity induced by concurrent chemoradiotherapy in patients with rectal cancer.Methods:A total of 115 patients with rectal cancer who underwent concurrent chemoradiotherapy in Zhejiang Cancer Hospital from March 2018 to August 2019 were prospectively selected. Nutritional risk was assessed by NRS 2002 and PG-SGA nutritional screening tools before, during and after radiotherapy. The acute toxicity was assessed by RTOG and CTCAE 3.0 scoring criteria. The correlation between nutritional status and the acute toxicity of chemoradiotherapy was analyzed by Spearman′ s correlation analysis. Results:The nutritional risk of the cohort was gradually increased from the beginning of chemoradiotherapy to the fourth week of chemoradiotherapy, and then decreased gradually. Spearman′ s correlation analysis showed that NRS 2002 and PG-SGA scores were positively correlated with acute hematological toxicity ( r=0.26, P<0.05; r=0.31, P<0.01), upper gastrointestinal toxicity ( r=0.51, P<0.01; r=0.63, P<0.01), proctitis ( r=0.23, P<0.05; r=0.45, P<0.01) and fatigue ( r=0.47, P<0.01; r=0.64, P<0.01) in patients with rectal cancer undergoing chemoradiotherapy. The correlation coefficients between PG-SGA and various toxicities were higher than those of NRS 2002. Stratified analysis showed that patients with stage Ⅱ-Ⅲ B, age<65 years and postoperative adjuvant chemoradiotherapy, nutritional status was significantly associated with the severity of toxicity (all P<0.05). Conclusions:Patients with rectal cancer has a high risk of malnutrition during concurrent chemoradiotherapy. The higher the risk of malnutrition, the greater the acute toxicity of chemoradiotherapy. Therefore, dynamic nutrition assessment and nutritional support should be strengthened for rectal cancer patients during chemoradiotherapy.