Objective To investigate the effect of Shenfu injectio on insulin resistance in rabbits undergoing cardiopulmonary bypass (CPB) . Methods Thirty New Zealand white rabbits of both sexes weighing 2.2-2.5 kg were randomly divided into 3 groups (n = 10 each): group I sham operation (group S); group Ⅰ CPB and group Ⅰ CPB + Shenfu injectio (group SFI). In group IE (group SFI) Shenfii injectio 10 ml/kg was injected intraperitoneally (IP) once a day for 2 days and at 30 min before anesthesia. In group Ⅱ and Ⅲ the duration between aortic cross-clamping and unclamping was 60 min. MAP was maintained at 50-60 mm Hg during CPB. The plasma glucose and insulin concentrations were measured immediately after induction of anesthesia (T_1), immediately after aortic cross-clamping (T_2) , and at 5, 35 and 75 min after aortic unclamping (T_3, T_4 , T_5) and insulin resistance index (HOMA-IR) was calculated. The expression of insulin receptor substrate-1 (IRS-1), phosphatidyl-inositol 3 kinase (PI3Kp85) and glucose transporters (CLUT4) in skeletal muscle was detected at 150 min of CPB by immuno-histochemistry. Results Compared with the baseline values at T_1 , plasma glucose and insulin concentrations and HOMA-IR were significantly increased at T_(2-5) in all 3 groups ( P < 0.05) . CPB significantly increased the plasma glucose and insulin concentrations and HOMA-IR at T_(2-5) in group Ⅱ and Ⅲ as compared with group S ( P < 0.05). The expression of PI3Kp85 and GLUT4 in muscle was significantly decreased in group Ⅱ and Ⅲ as compared with group S ( P < 0.05). Shenfu injectio significantly decreased plasma glucose concentration and HOMA-IR and increased plasma insulin concentration in group fD as compared with group Ⅱ . Shenfu injectio significantly attenuated the CPB-induced changes in IRS-1, PI3Kp85 and GLUT4 expression in muscle. Conclusion Shenfu injectio can ameliorate CPB-induced insulin resistance and the mechanism may be related to the up-regulation of the expression of insulin signal transduction molecules in skeletal muscle.

Shanghai University of TCM was the first Chinese medicine university that established field-grade experimental teaching center in China.And both of the Chinese medicine and Chinese herbs experimental teaching centers became national experimental teaching demonstration centers.There is general improvement in laboratories conditions,so the experimental teaching team building is the critical factor of improving experimental teaching quality.The experimental teaching team of Shanghai University of TCM consists of excellent teachers as its backbone,and lecturers and technicians from fields in traditional Chinese medicine Chinese herbs and clinical practices.The team members cooperate with each other by setting up experimental teaching research groups to improve teaching quality,which plays an important role in building experimental teaching demonstration center.

Objective To investigate the role of norepinephrine in the down-regulated visceral sensitivity of rats deprived of rapid eye movement(REM)sleep.Methods Twenty-four male Sprague- Dawley rats were randomly divided into 3 groups:cage-yoked rats as control(YC),rats with REM sleep deprivation(SD)and rats with yohimbine administered intraperitoneally after REM sleep deprivation(YSD).Flower pot technique was employed to make sleep deprivation model.YSD group was given vohimbine intraperitoneally at the 48th hour after REM sleep deprivation.After both SD and YSD groups had completed these processes,rats of all the three groups were given colorectal distension(CRD)and electromyogram (EMG)was recorded at the same time.The number of discharges of EMG and the threshold of Dain perception of the rats were observed to evaluate the visceral sensitivity.The thalamus,rectum and distal colon were taken after CRD;MAO-mRNA and TH-mRNA in three tissues were detected with RT-PCR.Resuits On 48th hour,the number of discharges of EMG in 10 seconds responding to CRD in group SD was significantly less than that in group YC and the threshold of pain perception in group SD was higher than that in group YC(P＜0.05).The number of discharge of EMG in group YSD was significantly more than that in group SD(P＜0.05).The expression of MAO-mRNA in group SD was lower than that in group YC(P＜0.05)and the expression of TH-mRNA in group SD was higher than that in group YC(P＜ 0.05).Conclusions The visceral sensitivity in rats is down-regulated by REM sleep deprivation,which can increase synthesis of norepinephrine.Norepinephrine can modulate visceral sensitivity.

OBJECTIVETo obtain recombinant human CYP2E1 and to determine its activity by using the specific probe substrate.

METHODSCYP2E1 cDNA was obtained by RT-PCR using human liver RNA as template. The cloned CYP2E1 cDNA was ligated with pFastBac vector to generate recombinant pFastBac-CYP2E1, which was then transformed into E. coli DH 10 Bac. Recombinant Bacmid-CYP2E1 was generated by transposition. Then Spodoptera frugiperda (Sf9) insect cells was infected with Bacmid-CYP2E1 to generate recombinant baculoviruses carrying human CYP2E1 cDNA. Finally, Sf9 insect cells were triinfected with recombinant baculoviruses carrying human CYP2E1, CYPOR and CYPb5. The activity of the recombinant enzymes was determined using chlorzoxazone as the substrate.

RESULTThe Kmand Vmaxof recombinant CYP2E1 to chlorzoxazone was (72.4 +/-8.7) micromol. L(-1) and (2.41 +/-0.10) micromol.min(-1)?g(-1)protein, respectively.

CONCLUSIONActive recombinant CYP2E1 has been obtained by bac-to-bac expression system and its activity is similar to previous reports.

Animals ; Baculoviridae ; genetics ; metabolism ; Cytochrome P-450 CYP2E1 ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Spodoptera ; genetics ; metabolism ; Transfection

BACKGROUND & OBJECTIVE: Recent studies showed overexpression of bcl-xL in human nasopharyngeal carcinoma (NPC) cell line CNE-2Z; it may play a pivotal role in tumorigenesis, metastasis, and drug resistance of NPC. This study was to explore inducing effect of bcl-xL short hairpin RNA (shRNA) on apoptosis of CNE-2Z cells. METHODS: After transfection of recombinant plasmid pmU6-RNAi expressing bcl-xL shRNA,apoptotic CNE-2Z cells were detected by fluorescent staining and flow cytometry (FCM). mRNA levels of bcl-xL, bcl-2, survivin, and caspase-3 was detected by reverse transcription-polymerase chain reaction (RT-PCR); while protein levels of Bcl-xL, Caspase-3, and P53 were detected by Western blot.RESULTS: When treated with pmU6-RNAi for 24 h, an obvious apoptotic peak of CNE-2Z cells appeared; cell shrinkage, chromatin condensation, and nuclear fragmentation were observed in most cells under fluorescent microscope. RT-PCR analysis showed that pmU6-RNAi down-regulated mRNA levels of bcl-xL, bcl-2, and caspase-3, but had little or no effect on mRNA level of survivin; Western blot analysis showed an obvious reduction in protein levels of Bcl-xL and Caspase-3, and a great increase in protein level of P53.CONCLUSIONS: bcl-xL shRNA can induce apoptosis of CNE-2Z cells, which may be closely related to down-regulation of bcl-2, caspase-3 and p53. bclxL shRNA may be helpful for developing gene therapy for NPC.

Objective To explore the method of fabricating larynx-shape tissue engineered cartilage by means of filling together with wrapping with pedicle myofascial flap.Methods Serial steps of solution casting,extrusion molding and particulate leaching were used to make larynx-shape[poly(3-hydroxybutyrate-co-3-hydroxyhexanoate),PHBHH]biomaterial models.The chondrocytes were seeded onto PHBHH models to form cell-PHBHH composites for culture in vitro for one week and then to fill and wrap larynx-shape composites with pedicle myofascial flap.After that to implant larynx-shape composites in situ on the back of adult New Zealan white rabbits(experimental groups n =9).Control groups(n =3)were the same measure as experimental groups but without chondrocytes on PHBHH models.Finally,morphological observation,HE & special staining and immunohistochemical test were conducted to evaluate the cartilage regeneration and its shape at 6,8 and 12 weeks following implantation.Results The PHBHH models appeared to be hollow half-trumpet with edges and comers of larynx-shape and its porosity ＞ 90％.Pedicle myofascial flap using fascia as lining presented rich blood supply and had enough to fill and wrap larynxshape composites.Tissue engineered larynx-shape cartilage specimens could be harvested at different period.It was demonstrated that the cartilaginous tissue formed in 6 weeks after implantation through histological and immunohistochemical examination and further maturity in 12 weeks and 18 weeks.But no cartilaginous tissue showed without chondrocytes on PHBHH as control groups to implant at the same time.Conclusion It seems that pedicled myofascial flap showed sufficient blood supply and that the filling together with wrapping method with pedicled myofascial flap is appropriate for fabricating larynx-shape tissue engineered cartilage.

OBJECTIVETo investigate the molecular genetic mechanism of a Chinese patient with mucopolysaccharidosis type I (MPS I).

METHODSPCR-sequencing analysis was applied to detect the mutations in exons in alpha-L-iduronidase gene (IDUA) of the patient. Restriction fragment length polymorphism (RFLP) and allele-specific oligonucleotide hybridization (ASO) were used to confirm the identified mutations. PCR amplified DNA samples from 50 normal individuals were sequenced to demonstrate that the newly identified mutation was not polymorphism.

RESULTSThe patient was compound heterozygous for a previously reported nonsense mutation Q60X (178C > T) in exon 2, inherited from the mother, and a newly detected missense mutation D203N (607G > A) in exon 6 from the father. The newly identified mutation D203N was not found in PCR amplified products from 50 normal individuals, indicating that it was not polymorphism.

CONCLUSIONThe two identified mutations may be the cause resulting in patient's clinical phenotype.

Adolescent ; Base Sequence ; Codon, Nonsense ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Humans ; Iduronidase ; genetics ; Male ; Mucopolysaccharidosis I ; genetics ; Mutation ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

BACKGROUNDThis is a prospective clinical study based on a large sample gathered from multiple centers in China, subordinating to 10th Five-Year Plan of National Science & Technology Progression. We analyzed the high-risk factors inducing hypertensive disorders in pregnancy (HDP) and estimated the potential effect of anti-oxidants administration, including vitamin C (VC), vitamin E (VE) and Salvia Miltiorrhiza L (SML), a Chinese herb medicine, in amelioration of the high-risk factors in pregnancy.

METHODSFrom April 2005 to July 2006, 4814 pregnant women from 24 national wide cooperative hospitals were involved in this prospective research. The participants were randomly divided into two groups: 1607 cases were in anti-oxidants group with administration of vitamins and SML; 3207 cases were in control group without any medicine given. Every participant was under monitoring for the morbidity of HDP and the high-risk factors were investigated in HDP cases in each group.

RESULTS(1) The morbidity of HDP was 3.55% in anti-oxidants group vs. 4.18% in control group. No statistical difference existed between the two groups (P > 0.05). (2) In anti-oxidants group, the HDP morbidities among three subgroups: VC + VE + SML, VC + VE and SML only, were 5.51%, 3.05% and 5% respectively. It showed no statistical difference among three remedies (P > 0.05). (3) The related index of factors affecting HDP showed in intensity sequence as follows: family HDP history > profession > education level > age > body weight. The incidence of HDP in normal population was 3.51%, and the incidence of HDP in high-risk pregnant women (family HDP history, heavy physical labor, low education level (middle school and below), age ≥ 40, body mass index ≥ 24) was 5.84%, which was obviously higher than that in normal population (P < 0.01). In anti-oxidants group, the probability of HDP in women with high-risk factors was 3.81%, which was obviously lower than that in control group with high-risk factors at 7.14% (P < 0.01). (4) In control group, the morbidity of HDP in women with family HDP history (especially with sisters'), heavy physical labor, middle school and below, age ≥ 35 was: 50.00%, 15.22%, 6.33%, 26.28% and 5.75%, respectively, and that in anti-oxidants group was 0, 7.69%, 3.74%, 9.27% and 2.67%, respectively, which was obviously lower than that in control group.

CONCLUSIONSThe high-risk factors prone to induce HDP included: family history of HDP, heavy physical labor, low education level, aging and obesity. No impressive effect of anti-oxidants application was found in preventing HDP in general population but the remedy demonstrated positive effect on preventing HDP in pregnant women with high-risk factors.

Adolescent ; Adult ; Age Factors ; Antioxidants ; therapeutic use ; Body Weight ; Female ; Humans ; Hypertension ; drug therapy ; physiopathology ; Logistic Models ; Pregnancy ; Pregnancy Complications ; drug therapy ; Prospective Studies ; Risk Factors ; Young Adult

Sclerosing angiomatoid nodular transformation (SANT) of the spleen, a newly defined primary lesion of the spleen, is very rare. Immunohistochemistry is the only way to confirm the diagnosis of SANT. We present the clinical characteristics and postoperative outcomes of five SANT cases that underwent splenectomy from January 2007 to October 2010. Although SANT had specific imaging findings, differential diagnosis from other splenic tuomrs or malignant lesions preoperatively was difficult. The hand-assisted laparoscopic splenectomy was a useful and effective technique for the management and postoperative diagnosis of SANT. All SANT patients had good prognosis without recurrence after splenectomy.
Adult ; Angiomatosis ; diagnosis ; pathology ; surgery ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Spleen ; pathology ; surgery ; Splenic Neoplasms ; diagnosis ; pathology ; surgery

Electronic Health Records ; Hospital Information Systems ; Hospitals ; Humans