Objective To investigate the expression of corticotropin-releasing factor (CRF) and its receptors including CRFR1 and CRFR2 on mouse mesenteric lymph nodes dendritic cells (MLNDC), and to analyze their effects on the biological phenotypes of intestinal dendritic cells .Methods The MLNDCs were isolated from C57BL/6 mice by using magnetic bead sorting .The purity of CD11c+DCs was identified by flow cytometry .The double-labeling immunofluorescence and the reverse transcriptase-polymerase chain reaction (RT-PCR) were performed to detect the expression of CRF , CRFR1 and CRFR2 on MLNDCs.The MLNDCs were exposed to CRF with or without the interference of CRFR 1 and CRFR2 antagonists .Flow cy-tometry was used to measure the changes of surface molecules ( MHCⅠ and MHCⅡ) and co-stimulatory molecules (CD80 and CD86).Results The CD11c+DCs accounted for (80.12±6.34)% of the isolate cells with a high cell viability of more than 90%.The expression of CRF , CRFR1 and CRFR2 at mRNA lev-el were detected in MLNDCs by RT-PCR.Results of the immunofluorescent staining assay indicated that both CRFR1 and CRFR2 were expressed on the surface of MLNDCs .The expression of CD86 on MLNDCs was inhibited by the treatment of MLNDCs with CRFR 1 antagonist , but enhanced by the treatment with CRFR2 antagonist .Conclusion Both CRF and CRFRs were detected in the MLNDCs isolated from the C57BL/6 mice.The CRF could alter the biological phenotypes of MLNDCs through binding to different CRFRs (CRFR1 and CRFR2), which affected the phenotypes of MLNDCs in opposite ways .

Objective To investigate the effect of local injection of Botulinum toxin type A(BTXA) on spasticity and function of the affected upper limb in stroke patients.Methods A total of 32 stroke patients were re- cruited and randomly divided into two groups:a BTXA group and a control group.All the patients had spasticity of upper limb muscles,which scored grade 2 to 3 with the Modified Ashworth Scale(MAS) ,and decreased elbow joint range of motion.The 16 patients in the BTXA group received BTXA injection in the biceps brachii muscles and flexor muscles of forearm on 10～15 points,while those in the control group did not.All the patients in both groups were treated with rehabilitation training techniques.The MAS,Fugl-Meyer upper limb function assessment and Barthel In- dex were employed to evaluate the changes of muscle tone,upper limb function and activity of living (ADL)perform- ance of the patients before injection and at 1st,2nd,6th 12th weeks after injection.Results The therapeutic effect between the BTXA group anti control group was significantly different in terms of biceps muscle tone,the scores of Fugl-Meyer upper limb function assessment and Barthel Index.Compared with preinjection,muscle tone was de- creased significantly and ADL performance was improved after injection in BTXA group.The effects of BTXA lasted more than 12 weeks.Conclusion Intramuscular muhipoint injection of BTXA was useful in reducing muscle spas- ticity,and was helpful for increasing motor ability of the affected upper limb and ADL performance of the stroke pa- tients.

The effectiveness and safety of acupuncture-moxibustion for the treatment of agrypnia was systematically reviewed. The clinical randomized controlled trial (RCT) of acupuncture-moxibustion for agrypnia were collected. The literature and document on acupuncture-moxibustion RCT for the treatment of agrypnia that published from January of 2001 to March of 2012 was searched with computer in PubMed, CNKI, Wanfang and VIP database. According to Cochrane Handbook 5. 1. 0, the bias risk and quality assessment were performed on every included trial and RevMan 5. 0 software was applied to make the Meta analysis. Totally 14 researches were included, involving 2662 cases. The Meta analysis showed that the differences of total effective rate between acupuncture-moxibustion and clinical routine treatment were significant [RR = 1.16, 95% CI (1.12, 1.19), Z = 9.32, P < 0.000 01]. The results indicate that total effective rate of acupuncture-moxibustion for agrypnia is obviously superior to that of clinical routine treatment, meaning clinical efficacy of acupuncture-moxibustion is served credit.
Acupuncture Therapy ; Amblyopia ; therapy ; Child ; Child, Preschool ; Female ; Humans ; Male ; Moxibustion

Objective To examine how stroke affects muscle coordination and whether muscle function will be improved after rehabilitation.Methods Ten stroke patients with mild hemiparesis and six age-and sex-matched controls were investigated at baseline.Velocity-encoded phase-contrast magnetic resonance imaging (VE-PC MRI) and surface eleetromyography (sEMG) were performed to evaluate muscle coordination of thigh muscles during knee extension and flexion and the effect of rehabilitation. Results Using VE-PC MRI,we found that the peak velocity of rectus femoris was lower in the affected limb (P

Based on the sequence of BAC (Bacterial Artificial Chromosome) along with the Cre/lox P system, the gene-targeting vectors to multiple loci of the repetitive internal transcribed spacers between rDNA genes in Leghorn chicken were constructed. The key material of multiple loci gene targeting in vivo would be obtained. First, the plasmid of pYLSV-TDN with TK, HRDS2, and Neo genes was constructed. The TK-HRDS2-Neo DNA fragment obtained from the plasmid of pYLSV-TDN was digested by Not I/HindIII and inserted into the upstream of the lox P site of BAC plasmid for obtaining the selective vector of BAC-TDN. The expression vector of pYLVS-GID with EGFP, hIFN genes, and HRDS1 was then obtained. The plasmid of BAC-TDN-VS-GID was obtained by cotransformation of the selective vector of BAC-TDN and the expression vector of pYLVS-GID to E. coli NS3529 through the action of Cre/lox P system. The gene-targeting vector of BAC-TDN-GID to multiple loci of the ITS region in Leghorn chicken was obtained by cleaving the sequence of pYLVS with the homing endonuclease of I -Sce I and ligating with the linker of LS. The insertion and the insert direction of DNA fragments were identified by restriction digestion or PCR and sequencing in each clone. The significance of the technique ofgene-targeting vector to multiple loci are shown as follows. First, the targeting loci were increased to 100 - 300. Second, the problems of unstable expression of inserted genes were partially solved. Third, the need for safety against toxicity integration was resolved. Fourth, the forbidden zone of gene integrating on the repetitive DNA sequences was broken through.
Animals ; Attachment Sites, Microbiological ; genetics ; Chromosomes, Artificial, Bacterial ; genetics ; Cloning, Molecular ; DNA ; genetics ; metabolism ; DNA Restriction Enzymes ; metabolism ; DNA, Ribosomal Spacer ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Integrases ; genetics ; Interferon-gamma ; genetics ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; Transformation, Genetic

Female ; Humans ; Manipulation, Orthopedic ; Middle Aged ; Radius Fractures ; surgery ; Ulna Fractures ; surgery ; Ulnar Nerve Compression Syndromes ; surgery

OBJECTIVETo prepare Form A and Form B of benazepril hydrochloride and to compare the differences in spectrums, thermodynamics and crystal structure between two polymorphic forms.

METHODSForm A and Form B of benazepril hydrochloride were characterized by Fourier transform infrared spectroscopy (IR), thermal gravimetric analysis (TG), differential scanning calorimetry (DSC), powder x-ray diffraction (PXRD) and single crystal x-ray diffraction (SCXRD).

RESULTSPreparation method, crystal structure and polymorphic stability of Form A and Form B of benazepril hydrochloride were obtained. Based on the analysis of crystal structure of both polymorphs, Form A belonged to monoclone space group P2(1) with a=7.8655(4)Å, b= 11.7700(6)Å, c= 13.5560(7)Å, β= 102.9470(10)°, V=1223.07 (11)Å(3) and Z=2, while Form B belonged to orthorhombic space group P212121, with a=7.9353(8)Å, b=11.6654(11)Å, c=26.6453(16)Å, V=2466.5(4)Å(3) and Z=4. From the DSC and XRD results, Form B of benazepril hydrochloride could be transformed into Form A after heating treatment.

CONCLUSIONForm A and Form B of benazepril hydrochloride are both anhydrous and displayed different polymorphs due to different molecular configuration. Furthermore, Form A exhibits more stable than Form B at high temperatures.

Benzazepines ; chemistry ; Crystallization ; Drug Stability ; Molecular Conformation

OBJECTIVETo summarize the phenotypes and identify SCN1A mutations in families with generalized epilepsy with febrile seizures plus (GEFS(+)), and analyze the genotype- phenotype correlations in GEFS(+) families.

METHODGenomic DNA was extracted from peripheral blood lymphocytes of the proband and other available members in the GEFS(+) families. The phenotypes of the affected members were analyzed. The coding regions and flanking intronic regions of the SCN1A gene were screened for mutations using PCR and direct DNA sequencing.

RESULTIn 39 GEFS(+) families, there were 196 affected members, ranging from 2 to 22 affected members in each family. Their phenotypes included febrile seizures (FS) in 92(46.9%), febrile seizures plus (FS(+)) in 62(31.6%), FS or FS(+) with partial seizures in 12(6.1%), afebrile generalized tonic-clonic seizures (AGTCS) in 11(5.6%), myoclonic atonic epilepsy in 8(4.1%), Dravet syndrome in 2(1.0%), childhood absence epilepsy in 1 (0.5%), FS(+) with myoclonic seizures in 1(0.5%), AGTCS and myoclonic seizures in 1 (0.5%), partial seizures in 1 (0.5%), unclassified seizures in 5 (2.6%). Four families were found with SCN1A mutations, including three families with missense mutation (N935H, R101Q, G1382R) and one family with truncation mutation (C373fsx378). In three families with missense mutations, the phenotypes include FS, FS(+), FS(+) with partial seizures, and AGTCS. In one family with truncation mutation, the phenotypes included FS, FS(+), and Dravet syndrome. The mother of proband in the family with missense mutation (R101Q) and the father of proband in the family with truncation mutation (C373fsx378) were both somatic mosaicism. Both of their phenotypes were FS(+).

CONCLUSIONThe most common phenotypes of GEFS(+) were FS and FS(+), followed by the FS/FS(+) with partial seizures and AGTCS. The most severe phenotype was Dravet syndrome. SCN1A mutation rate in GEFS(+) was about 10%. Missense mutation was common in GEFS(+) families, few with truncation mutation. Few members of GEFS(+) families had somatic mosaicism of SCN1A mutations and their phenotypes were relatively mild.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Epilepsies, Myoclonic ; genetics ; Epilepsy, Generalized ; genetics ; Female ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; genetics ; Mutation, Missense ; NAV1.1 Voltage-Gated Sodium Channel ; genetics ; Pedigree ; Phenotype ; Seizures, Febrile ; genetics

Objective To investigate the application value of lymph node-targeted chemotherapy in resection of esophageal carcinoma.Methods The prospective study was conducted.The clinical data of 117 patients with middle and low esophageal carcinoma (without involving esophagogastric junction) who underwent left transthoracic esophagectomy and regional lymph node dissection in the West China Hospital of Sichuan University between January 2013 and December 2013 were collected.All patients were allocated into the 4 groups by semi-randomized control method:CPL group received intraoperatively carbon nanoparticles-paclitaxel for lymph node-targeted chemotherapy,CFL group received intraoperatively carbon nanoparticles-fluorouracil for lymph nodetargeted chemotherapy,FV group received preoperative fluorouracil intravenous chemotherapy,and control group underwent left transthoracic esophagectomy and regional lymph node dissection.CPL and CFL groups:suspensions of carbon nanoparticles and chemotherapy drugs were preoperatively prepared,and were intraoperatively injected under mucosa of lower edge of thoracic esophageal tumor using 0.1 mL syringes.FV group:fluorouracil with 100 mL of saline were mixed,and then were preoperatively injected by intravenous drip within 30 minutes.After cutting tissues of esophageal carcinoma,lymph nodes of left gastric arteria were removed,and drug level in lymph nodes was measured.At the beginning of esophagogastrostomy in the CPL,CFL and FV groups,3 mL peripheral venous blood were collected and measured for serum drug level.Control group:patients underwent left transthoractic esophagectomy and regional lymph node dissection (no blood sample and esophageal specimen).Observation indicators:(1) comparison of drug levels in lymph node and serum of patients with chemotherapy;(2) follow-up and survival:4-year cumulative survival rate in 4 groups.Follow-up using outpatient examination and telephone interview was performed to detect patients' survival up to December 2017.Measurement data with normal distribution and homogeneity of variance were represented as (x)±s,and comparisons among groups were analyzed using the ANOVA.Measurement data with skewed distribution were described as M (P25,P75),and comparisons among groups and between groups were respectively analyzed using the Kruskal-Wallis rank test and Mann-Whitney U test or Wilcoxon signed rank test.Comparisons of count data were analyzed using chi-square test or Fisher exact probability.Ordinal data were compared by the Kruskal-Wallis test.The survival curve was drawn by the Kaplan-Meier method,and Log-rank test was used for survival analysis.Results One hundred and seventeen patients were screened for eligibility,including 90 males and 27 females,and age was 37-84 years old,with an average age of 62 years old.Of 117 patients,41,41,9 and 26 were respectively allocated into the CPL,CFL,FV and control groups.Eligible patients recovered and were discharged from hospital,without bone marrow depression,severe diarrhea,anastomotic leakage and severe pneumonia.(1) Comparison of drug levels in lymph node and serum of patients with chemotherapy:drug levels in the CPL,CFL and FV groups were respectively 2.16 μg/g (1.14 μg/g,4.39 μg/g),0.44 μg/g (0.11 μg/g,1.18 μg/g),0.11 μg/g (0,0.28 μg/g) in lymph nodes and 0 (0,0),0 (0,0.31 μg/mL),0 (0,0.30 μg/mL) in serum.Drug levels of lymph node in the CPL and CFL groups were higher than those of serum,with statistically significant differences (Z=-5.579,-3.069,P＜0.05).There was no statistically significant difference in drug levels of lymph node and serum of FV group (Z =-0.365,P＞0.05).There was a statistically significant difference in drug levels of lymph node among CPL,CFL and FV groups (H=33.458,P＜0.05),and in drug levels of serum among CPL,CFL and FV groups (H=10.356,P＜0.05).Further analysis showed that fluorouracil level of lymph node in the CFL group was higher than that in the FV group,with a statistically significant difference (Z =82.500,P＜ 0.05),and there was no statistically significant difference in fluorouracil level of serum between CFL group and FV group (Z =160.500,P＞0.05).Paclitaxel level of lymph node in the CPL group was higher than fluorouracil level of lymph node in the CFL group,with a statistically significant difference (Z =351.000,P＜0.05),and paclitaxel level of serum in the CPL group was lower than fluorouracil level of serum in the CFL group,showing a statistically significant difference (Z=577.000,P＜0.05).(2) Follow-up and survival:of 117 patients,21 lost follow-up,and 96 were followed up for 6.0-58.0 months,with a median time of 20.0 months.The 4-year cumulative survival rate in the CPL,CFL,FV and control groups was respectively 46.2％,27.8％,33.3％ and 17.1％.There was no statistically significant difference in the survival of 4 groups (x2 =5.166,P＞0.05).Conclusions The lymph node-targeted chemotherapy can promote chemotherapy drugs to aggregate in the lymph nodes during resection of esophageal carcinoma.The affinity of chemotherapy drugs on carrier is involved in clinical effects,and single use of chemotherapy drug cannot improve postoperative survival rate of patients.