Objective To evaluate the effects of butorphanol pretreatment on myocardial damage induced by limb ischemia/reperfusion (I/R) in rats.Methods One hundred and eight male Sprague-Dawley rats,weighing 180-220 g,were randomly divided into 3 groups (n =36 each) using a random number table:sham operation group (group S),I/R group and butorphanol pretreatment group (group B).Limb ischemia was induced by occlusion of bilateral hind limbs for 2 h followed by reperfusion in I/R and B groups.At 10 min before ischemia,butorphanol 0.2 mg/kg was injected via the right jugular vein in group B,while the equal volume of normal saline was injected in group I/R.Six rats were chosen randomly before ischemia (baseline,T0) and at 2,6,12,24 and 48 h after reperfusion (T1-5) and blood samples were taken from the abdominal aorta for determination of the serum creatine kinase isoenzyme-MB (CK-MB) activity and cardiac troponin Ⅰ (cTnI) concentration.The animals were then sacrificed and myocardial specimens were obtained for determination of myocardial apoptosis (using TUNEL) and the expression of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated protein kinase (ERK) (using Western blot analysis).Apoptosis index was calculated.Results Compared with group S,the serum CK-MB and cTnI levels were significantly increased at T1-5,and the apoptosis index and expression of JNK and ERK were increased at T2-5 in group I/R (P ＜ 0.05).Compared with group I/R,the pathological changes of myocardium were significantly reduced at T4,the serum CK-MB and cTnI levels were decreased at T2-5,the apoptosis index was decreased at T4,5,the expression of ERK was up-regulated at T2-5,and the expression of JNK was down-regulated at T2-4 (P ＜ 0.05).Conclusion Butorphanol pretreatment can reduce apoptosis in myocardial cells through activating MAPK signal transduction pathway,up-regulating ERK expression and down-regulating JNK expression,thus attenuating myocardial damage induced by limb I/R in rats.

Objective To clone,express and characterize a tegument protein gene of Schistosoma japonicum(Sj29),and investigate the immune protection of the recombinant protein against S.japonicum in mice.Methods The gene coding for Sj29 protein was amplified by PCR,and the sequence was analyzed by bioinformatics tools.Partial fragment of Sj29 gene was subcloned into the prokaryotic expression vector pET28c(+).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced the recombinant with IPTG.The recombinant protein(rSj29) was purified by His-binding-resin affinity chromatography and characterized by Western blotting.Three groups each with 10 BALB/c mice were immunized subcutaneously three times(two weeks interval) respectively with 100 ?l recombinant rSj29(0.1 mg/ml),adjuvant or PBS.At the 15th day after the final inoculation,each mouse was challenged by 40 ?2 cercariae of S.japonicum.At the 53th day after infection,the mice were sacrificed to obtain the number of adult worms,number of eggs in liver and feces.Serum samples were collected at pre-immunization and certain time after immuniza-tion,and were analyzed for IgG by ELISA.The localization of rSj29 in worms of different developmental stages was demonstrated by immunofluorescent technique.mRNA expression level of Sj29 gene in worms of different developmental stages and three groups after infection was detected by quantitative real-time PCR.Results A 576 bp Sj29 gene fragment was obtained.The recombinant protein rSj29 with Mr 22 900 was expressed in the form of inclusion body.The recombinant rSj29 can be recognized by sera of mice immunized with rSj29 and sera of infected mice.The number of adult worms(15.4?5.9),number of hepatic eggs(40 143.3?2 995.9) and number of fecal eggs(3 803.9?110.9) in re-combinant protein group were significantly higher than those of PBS control group(20?3.4,49 318.1?6 648.3,5 238.1? 303.5,respectively)(P

Objective: To observe the controlled hypotension effects of nicardipine in 2 different ways for spinal tumor operalion. Methods: Twenty-four adult patients, scheduled for selective spinal tumor operation, were randomly divided into 2 groups. In groupⅠ(n=12), the nicardipine was infused at a rate of 10 μg*kg-1*min-1 and the infusion continued until MAP was at the level of 7.33-8.66 kPa, and then the rate was decreased to 1 μg*kg-1*min-1. In Group Ⅱ(n=12), nicardipine was given 0.01-0.02 mg/kg as the load dose, then infused at 1-2 μg*kg-1*min-1. Results: During the period of controlled hypotension, cardiac index(CI) increased significantly, other hemodynamic variables were stable and no hypertension rebound occurred in both groups. Reaching time of target blood pressure in groupⅡ was shorter than that in groupⅠ（P＜0.05）. The dose required to obtain target blood pressure in group Ⅱwas less than that in group Ⅰ（P＜0.05）. BP recovery time from discontinuing nicardipine infusion to pre-hypotension level,bleeding volume and transfusion volume were similar between 2 groups（P＞0.05）.During mass bleeding,　serious arrhythmia and oliguria did not occur in any case. Conclusion: Controlled hypotension with nicardipine is rapid, stable and easily controlled without hypertension rebound. Nicardipine has considerable protective effects on heart and kidney during mass bleeding. The method of bolus injection followed with intravenous infusion is more suitable to clinical application.

OBJECTIVETo study the effect and mechanism of action of norcantharidin on proliferation and invasion of GBC-SD cells.

METHODSGBC-SD cells of human gallbladder carcinoma were cultured by cell culture technique. The tetrazolium-based colorimetric assay was used to evaluate cell growth. The Matrigel experiment and the crossing-river test were used to examine the invasiveness of GBC-SD cells. Expression of MMP(2), TIMP(2), PCNA and Ki-67 proteins of GBC-SD cells was determined by streptavidin-biotin complex method.

RESULTSNorcantharidin inhibited the growth and proliferation of GBC-SD cells in a dose and time dependent manner, with an IC(50) value of 56.18 micro g/ml at 48 h. The Matrigel experiment showed that norcantharidin began to inhibit the in vitro invasion of GBC-SD cells at the concentration of 5 micro g/ml. At 40 micro g/ml, the invasive action of GBC-SD cells was inhibited completely and their crossing-river time was prolonged significantly. After treatment with norcantharidin, the expression of PCNA, Ki-67, MMP(2) was significantly decreased. With the increase in TIMP(2) expression, the MMP(2) to TIMP(2) ratio was decreased significantly (P < 0.05).

CONCLUSIONNorcantharidin inhibits the in vitro proliferation and growth of human gallbladder carcinoma cells at relatively low concentrations by inhibiting PCNA and Ki-67 expression. Its anti-invasive activity may be the results of decrease in MMP(2) to TIMP(2) ratio and reduced cell motility.

Antineoplastic Agents ; pharmacology ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Gallbladder Neoplasms ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Neoplasm Invasiveness ; Proliferating Cell Nuclear Antigen ; metabolism ; Time Factors ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

BACKGROUNDOn May 12, 2008, a major earthquake hit Wenchuan County in Sichuan Province of China. The number of cases of crush injury following this event was high. Ultrasonic appearance of rhabdomyolysis (RM) caused by crush injury in the Wenchuan earthquake was observed to evaluate the diagnostic value of ultrasound for detection of rhabdomyolysis.

METHODSWe analyzed clinical and ultrasonic manifestations of 50 cases of RM and 18 cases of RM with osteofascial compartment syndrome (OCS). All cases were caused by crush injury in the Wenchuan earthquake. For these RM patients, we also evaluated the correlations between creatine kinase (CK) and the scope of the muscle lesions as observed by ultrasound.

RESULTSThere were differences in clinical symptoms, physical signs and ultrasonic appearance between the two groups of patients. The ultrasonic characteristics of the RM were as follows: the striated muscle in the lesions thickened with good overall continuity, and the muscle texture was vague; the strength of the echo was uneven and the echo was cloudy or ground glass-like. Liquid dark zones appeared between muscles and were spindle-like or irregular in shape. There were no blood flow signals in the liquid dark areas. The volume of the striated muscle increased in patients with OCS; the fascia wrapping the muscle showed arched protrusions and significant displacement. The flow velocity of the distal arteries decreased and the spectrum was abnormal. The muscle lesion scope of RM group and RM and OCS group was (7.8 +/- 2.0) cm and (13.6 +/- 3.1) cm, respectively. The correlation coefficient (r) between the muscle lesion scope and the CK was 0.681 for the RM group (P < 0.05) and 0.516 for the RM and OCS group (P < 0.05).

CONCLUSIONSThe ultrasonogram of RM has characteristic manifestations and can provide important information for clinical diagnosis and treatment of rhabdomyolysis.

Adolescent ; Adult ; China ; Compartment Syndromes ; diagnostic imaging ; Crush Syndrome ; diagnostic imaging ; Earthquakes ; Female ; Humans ; Male ; Rhabdomyolysis ; diagnostic imaging ; Ultrasonography ; Young Adult

Objective To investigate the efficiency and safety of allogeneic hematopoietic cell transplantation for malignant hematological diseases in patients older than 50 yeas of age. Methods From May 2002 to January 2010, 35 patients (＞ 50 years) with malignant hematological diseases received allogeneic hematopoietic cell transplantation. In 35 patients, 18 patients were conditioned with non-myeloablative regimen and 17 patients with myeloablative regimen. The outcome,engraftment and prognosis of allogeneic hematopoietic cell transplantation were analyzed. Results The hematopoetic reconstitution was achieved in 32 of 35 patients. The median time of granulocyte count exceeding 0. 5 × 109/L was 12 days and the that of platelet count exceeding 20 × 109/L was 17days. The cumulative incidence of aGVHD was 48. 6 %, and 37. 9 % patients developed cGVHD.The estimate probability of cumulative survival at 5 years was 48. 5 %, The estimate probability of cumulative mortality rate was 51.5 %, and the estimated transplant-related mortality was 22. 9 %.The relapse rate was 11.4 %. There was significant difference except for the incidence of cGVHD.Conclusion Allogeneic hematopoietic cell transplantation may be appropriate for older patients with malignant hematological diseases.

OBJECTIVETo explore the anti-tumor mechanism of norcantharidin (NCTD) for the implanted tumors of human gallbladder carcinoma in nude mice in vivo.

METHODSAnimal model of implanted tumors of human gallbladder carcinoma in nude mice was established. Mice were randomly divided into control, 5-FU, NCTD and NCTD + 5-FU groups and were taken different treatment. The expressions of PCNA, Ki-67, cyclin D1, p27, Bcl-2, Bax, Survivin, nm23/nm23-H1, MMP2 and TIMP2 proteins or genes in each tissue section of every group were determined by immunohistochemistry and RT-PCR.

RESULTS(1) On proliferation-related gene proteins, the expression of PCNA, Ki-67, cyclin D1 was significantly decreased, with significantly increased expression of p27 protein, in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of PCNA mRNA, cyclin D1 mRNA was decreased, with significantly increased expression of p27 mRNA in NCTD group. (2) On apoptosis-related gene proteins, the expression of Bcl-2 was significantly decreased in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of Bcl-2 mRNA, Survivin mRNA was significantly decreased, with significantly increased expression of Bax mRNA in NCTD group. (3) There was significant difference on invasion around tumor and lung metastasis in NCTD group when compared with control group (P < 0.01). On metastasis-related gene proteins, the expression of nm23 and TIMP2 was significantly increased, with significantly decreased expression of MMP2 in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of nm23-H1 mRNA, TIMP2 mRNA was significantly increased, with significantly decreased expression of MMP2 mRNA in NCTD group.

CONCLUSIONSThe anti-tumor mechanism of NCTD for human gallbladder carcinoma in nude mice might correlated with inhibition of cell proliferation, blockage of cell cycle, induction of cell apoptosis, reducing of cell motility and invasive capability, alteration of the expression of proliferation-, apoptosis- and metastasis-related gene proteins such as PCNA, Ki-67, cyclin D1, p27, Bcl-2, Bax, Survivin, nm23, MMP2 and TIMP2.

Animals ; Apoptosis ; drug effects ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Proliferation ; drug effects ; Cyclin D1 ; biosynthesis ; genetics ; Gallbladder Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; bcl-2-Associated X Protein ; biosynthesis ; genetics

Objective To analyze the influence of validating the parental origin to the interpretation of clinical pathogenicity of total 54 copy number variations(CNV)with different clinical significance in 46 patients undergo chromosomal microarray analysis(CMA).Methods A retrospective study.This study enrolled 46 patients conducted in Department of Pediatric Endocrinology and Genetics of Shanghai Xinhua Hospital during the period of August 2014 to December 2015,involving 54 different CNVs detected by CMA.The parental origin of CNVs was examined by CMA or quantitative real-time polymerase chain reaction.Results Totally 54 different CNVs were found in 46 patients by CMA.Seventeen out of the 54 CNVs were pathogenic variations.After validating the parental origin,14 CNVs were proved de novo mutation,while 3 CNVs have maternal origin including 1q21.1 deletion syndrome,Xq27.3q28 and Xq22.1q22.3 duplications which inherited from maternal X chromosome.CNVs of 1q21.1 deletion syndrome often inherited from parents,and no phenotype appears on mother which may be due to the deactivation mechanism of duplications on mother′s X chromosome.Therefore,these 17 pathogenic variations were still considered to be clinical pathogenic significance after validating the parental origin.Ten out of 54 CNVs were variants of uncertain significance-likely pathogenic.After parental original validation,3 CNVs were proved de novo mutation considering likely pathogenic significance,while 7 CNVs have parental origin still judged to be unknown clinical pathogenicity.Twenty-seven out of 54 CNVs were variants of uncertain significance.After validating the parental origin,only 1 CNV was proved de novo mutation considering likely pathogenic significance,while all the others had parental origin considered to be variations likely benign.Conclusion CNVs reported as likely pathogenic should be validated the parental origin in order to further study their clinical pathogenicity,while variants of uncertain significance can preliminary clear its nature by validating parental origin.

OBJECTIVETo determine the classification of masticatory myospasm by analyzing characteristics of clinical appearances.

METHODSThirty-six cases of masticatory myospasm from 2000 to 2010 were included. The clinical data of these patients were analyzed, including patient information, patient history, clinical characteristics, severity and the frequency of myospasmodic movement, electromyogram (EMG), and the efficacy of botulinum toxin injection treatment.

RESULTSThere were 11 males and 25 females, aged from 15 to 71. According to the clinical manifestation and EMG findings, patients could be divided into two groups: 18 cases were classified as jaw closing type which involved masseter and/or temporalis muscles presenting as trismus and acute pain, the other 18 cases were jaw opening type which involved lateral pterygoid muscles complaining difficulty in jaw closing and teeth clenching. The jaw closing type was often seen in patients of 20 to 50 years old, the jaw opening was frequently seen in patients over 50 years old. Jaw closing type was attacked intermittently and unilaterally, but jaw opening was often attacked continually and bilaterally. The rating scale of the severity of spasmodic movement was not different between the two types, but the frequency of spasmodic attack was much higher for jaw opening type (P < 0.05). The EMG of jaw closing type was classified into persistent, rhythmic and irregular type. The EMG of jaw opening type was classified into spontaneous and exercise-induced type. Twelve cases were treated by botulinum toxin injection that could significantly relieve symptoms.

CONCLUSIONSMasticatory myospasm can be classified into jaw closing and jaw opening types. Jaw closing type involves masseter and/or temporalis muscles and jaw opening type involves lateral pterygoid muscles. Botulinum toxin injection was the most effective therapy for the masticatory myospasm.

Adolescent ; Adult ; Aged ; Botulinum Toxins, Type A ; therapeutic use ; Electromyography ; Female ; Humans ; Jaw ; physiopathology ; Male ; Masseter Muscle ; physiopathology ; Masticatory Muscles ; physiopathology ; Middle Aged ; Pterygoid Muscles ; physiopathology ; Spasm ; classification ; drug therapy ; physiopathology ; Temporal Muscle ; physiopathology ; Young Adult

[Objective] To investigate the purchase and use of edible oil , as well as recycling of kitchen waste oil in Shanghai catering service units , providing reference for relevant regulatory authorities .
[ Methods] A total of 976 units of various types of catering service in Shanghai were selected via strati-fied sampling methodology , a self-designed questionnaire used to investigate the purchase and use of edi-ble oil, as well as recycling of kitchen waste oil .Meanwhile, an on-site rapid detection technology was used to test items including acid value , peroxide value , polar components of unused pre -packaged edible oil and also the process of using frying oils on site . [ Results] The amount of edible oil used in catering service units of Shanghai was about 1235 tons per day;6.0% of the oil purchased was bulk cooking oil , which accounted for 5 .6%of the total amount consumed;44 .3%of the catering service units signed with professional companies for the recycling of kitchen waste oil;3615 samples were detected using an on -site rapid detection , of which 17 were suspected to be positive .The lab confirmed that test results were quali-fied. [ Conclusion] The edible oil used in Shanghai catering service units is safe in general , while bulk cooking oil may have potential safety hazards .Food practitioners have low awareness of health risk caused
by using edible oil repeatedly and for too long time , which makes some enterprise use “old oil” for too long;There does exist the possibility of kitchen waste oil returning to food chain as its recycling has not been standardized .