Objective:To clone interleukin 4 cDNA of Tibet pig.Methods:To amplify cDNA,using RT PCR from the total RNA from the lymphocyte stimulated by 10 ?g/ml Con A in vitro,to subclone into pMD T vector,and then to determine it by digesting and sequencing.Results:The cloned Tibet pig interleukin 4 was 98% homologus to the necleotide acids of Chenghua pig a special specie of pig in Sichuan province and 99% homologus to that of hybrid of Landrance with Chanbai pig.Conclusion:IL 4 of Tibet pig was successfully cloned from the lymphocyte after stimulating 70 h with ConA.

ObjectiveTo demonstrate the effect of moderate hypothermia and doule carotid ice applied to acute intracerebral hemorrhage (ICH) and the best opportunity of treatment.Methods174 cases with ICH were randomly divided into treated group and control group, 87 cases for each group. Each case of both groups was treated by routine therapy, moreover, all cases in treated group were treated by moderate hypothermia (MHT).The clinical effect and prognosis between treated group and control group were contrasted,so to do between the cases in the treated group who were treated no more than 3 hours and more than 3 hours after attack.ResultsCompared with the control group, the neurological function improved in the treated group(P<0.05), as well as survival rate and recovery(P<0.05).Cases who treated no more than 3 hours after attack was better than that of more than 3 hours(P<0.05).ConclusionModerate hypothermia can effectively ease the neurologic deficits after ICH, decrease mortality and disable rate.The best therapy time is within 3 hours after ICH.

Background Central corneal thickness (CCT)is an important parameter to evaluate corneal healthy status and is crucial for surgical planning.However,clinical study found that the center of cornea does not correspond to the thinnest point of cornea.Thus,it is essential to characterize the minimum corneal thickness(MCT) and its location.Objective Present study was to determine the thickness and location of MCT and its relationship to the fellow eye using Pentacam High Resolution technique.Methods The 564 eyes from 282 Chinese myopic patients were reviewed in this study.The CCT,MCT,pupillary central corneal thickness(PCT) and x-y coordinate of thinnest point were bilaterally measured.Written informed consent was obtained prior to the ocular biomedical measurement.Results CCT was (540.07±31.78) μm in the right eyes and (539.24±31.06) μm in the lefi eyes; PCT was (540.25±30.75)μm in the right eyes and (539.48±31.00)μm in the left eyes.MCT was (537.87± 31.91)μm in the right eyes and (536.35±31.24)μm in the left eyes,showing significant differences in all the parameters between the right eyes and left eyes expect for PCT(CCT:P=0.046;PCT:P=0.065 ;MCT:P=0.000).The C CT,PCT and M CT were significantly correlated between the right eyes and left eyes (r =0.97,0.97,0.98,P＜ 0.01).Bland-Altman plot showed a good consistence between the both eyes.The mean distance from the center was (0.50±0.21) mm in the right eyes and (0.56±0.22)mm in the left eyes,showing a significant difference (P =0.000).The difference between CCT and MCT was approximately (2.20±1.74)μm in the right eyes and (2.88±1.75) μm in the left eyes.The location of MCT in both the right eyes and left eyes tended to symmetry along the vertical midline.The distance between the R (x,y) to transformed L (x,y) was (0.29 ± 0.17)mm and the angular distance was (28.28±28.21)degree.Conclusions This study offers a range of MCT and its location in Chinese myopic patients.The difference exists between the CCT and MCT in bilateral eyes.The location of the thinnest point tends to be symmetrical along the vertical midline between the both eyes.The changes of these parameters may be helpful for the diagnosis of some corneal diseases.

OBJECTIVETo explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.

METHODSForty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.

RESULTSCompared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).

CONCLUSIONSEA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.

Animals ; Disease Models, Animal ; Electroacupuncture ; Ghrelin ; metabolism ; HMGB1 Protein ; metabolism ; Jejunum ; metabolism ; Male ; Rats ; Rats, Wistar ; Sepsis ; metabolism

0.05).There was significant difference in the 3-year survival rate between A and C group. Conclusions The 3-year survival rate was dramatically increased with combined therapy mainly by cisplatin, the dose of 60～80mg is tolerant for the elderly aged above seventy years, and perioperation complications can be cured.

Objective To explore the change of plasma orexin A concentration and the correlation between plasma orexin A concentration and energy intake in obese children.Methods Fasting plasma orexin A concentrations,boaly mass index(BMI) and energy intake were measured in 48 obese children(obese group) and 48 healthy children(healthy control group),and these indexes were compared,the correlation between plasma orexin A concentration and BMI,energy intake were analyzed.Results 1.The plasma orexin A concentration in obese group was significantly lower than that in healthy control group(F=5.632 P=0.008).2.In obsess group,there were negative correlation between plasma orexin A concentration and BMI(r=-0.478 P=0.012),positive correlation between plasma orexin A concentration and total energy intake(r=0.503 P=0.007),fat intake(r=0.659 P=0.006) and protein intake(r=0.381 P=0.026),and there was negative correlation between plasma orexin A concentration and carbohydrate(r=-0.316 P=0.022).3.In healthy control group,there were negative correlation between plasma orexin A concentration and BMI(r=-0.491 P=0.018),positive correlation between plasma orexin A concentration and total energy intake(r=0.512 P=0.009),fat intake(r=0.406 P=0.005),protein intake(r=0.313 P=0.020),and carbohydrate(r=0.432 P=0.025).Conclusions Orexin A may be involved in regulation of energy metabolism in obese children,and the interaction between plasma orexin A and energy intake might be different in different nutritional status in children.

[ABSTRACT]AIM:Toexploretheeffectoftheelasticmodulusandsizesofliquidcrystal(LC)phasesonosteo-genic differentiation based on OPC/PU composite substrate by mimicking the microenvironment in rat bone mesenchymal stem cells (rBMSCs).METHODS: A series of composite substrates with different elastic modulus were constructed via modulation of LC content in the composites .The surface phase structure was observed by polarized microscopy , and the mechanical property was measured by a universal material testing machine .Furthermore, the laser confocal microscope was employed to observe the spreading , polarization and the cytoskeleton arrangement of the rBMSCs .The proliferation of rBM-SCs was evaluated by CCK-8 assay.The specific mRNA expression of osteogenic differentiation such as collagen Ⅰ, and osteopontin on the composite membranes was detected by real-time PCR.RESULTS:The size and number of LC phase in-creased and the elastic modulus of the composite substrates decreased with the increase of the LC content .The rBMSCs ex-hibited better characteristics of initial adhesion , spreading and proliferation on the OPC 10-PU and OPC30-PU in the early and medium culturing .The rBMSCs displayed higher expression of collagen Ⅰ and osteopontin on the OPC10-PU in the early and medium osteogenic induction , while the high expression of these osteogenic genes occured on the OPC 30-PU and OPC50-PU in later osteogenic induction .The emphasis of genetic expression was switched from collagen Ⅰin the early and medium osteogenic induction to osteopontin in the later stage .CONCLUSION:When the content of LC remained low in the composite substrates , rBMSCs mainly responded to the mechanical stimuli induced by substrate stiffness and exhibited distinguished cellular behaviors;with the increase in the LC content , rBMSCs had strong interactions with LC by sensing the viscoelasticity of LC , probably resulted from the contribution of both substrate stiffness and the viscoelasticity of LC phase .

Objective To study the influence of electro-acupuncture(EA)at Zusanli points(足三里穴) on the apoptosis of thymocytes in rats with abdominal infection and its mechanism.Methods A total of 40 Sprague-Dawley(SD)rats were randomly divided into four groups,including normal control group,model group,non-acupoint group and Zusanli group.The abdominal infection model of rat was made by cecal ligation and puncture(CLP).After abdominal cavity infection for 36 hours,the apoptosis of thymocytes was observed under electron microscope and light microscope,and the apoptosis ratio of thymocytes was determined by Annexin V-PI method with flow cytometry technique.The content of Bcl-2 protein of thymocytes and concentration of corticosterone in plasma were determined.Results Abdominal infection resulted from CLP could significantly increase the apoptosis of thymocytes and lead to the typical histopathological changes of apoptosis of thymocytes under electron microscope and light microscope.Apoptosis ratios of thyrnocytes in model group[(44.7?3.3)%],non-acupoint group[(42.7?3.0)%]and Zusanli group[(32.6?3.3)%] were significantly higher than the ratio in the control group[(21.2?2.3)%,all P0.05).Abdominal infection resulted from CLP also could reduce the content of Bcl-2 protein of thymocytes.The content of Bcl-2 protein of thymocytes in model group(71.2?5.6),non-acupoint group(73.5?5.9)and Zusanli group(82.4?6.8) were significantly lower than normal control group(95.3?6.3,all P

OBJECTIVETo investigate the effects and mechanisms of diethylhexylphthalate (DEHP) on morphology and function of progenitor Leydig cells (PLC) in rats.

METHODSTwenty pregnant SD rats were randomly divided into 4 groups ( n = 5): normal control group, DEHP low dose group , middle dose group, and high dose group, which were treated from postnatal day (PND) 1 to PND 21 of the pubs with DEHP at the doses of 0, 10, 100, 750 mg/(kg · d) in 0.5 ml of corn oil by gavage respectively. At the end of the treatment, the male pups were killed and blood samples were collected for determination of serum testosterone concentration by chemiluminescence method. The body weight, testis weight and anogenital distance (AGD) were measured. The morphology of PLC was observed by light and transmission electron microscopy. The protein expression of steroidogenic acute regulatory protein(StAR) in PLC was determined by immunohistochemistry. The mRNA expression of insulin-like growth factor-I (IGF-I) in the testis was assayed by real-time PCR.

RESULTSCompared with normal control group, the serum testosterone and AGD of male pubs from the middle and high dose groups were declined significantly (P < 0.01), the testis weight and body weight from high dose group were decreased significantly (P < 0.01), while the testis weight increased in the low dose group (P < 0.05). Under light microscope, PLC showed hyperplasia and cluster aggregation in the low dose group and focal hyperplasia in the middle and high dose group. The spermatogenic cells in seminiferous tubules showed decrease, apoptosis and unfix in the high dose group. Under transmission electron microscope, the PLC showed decreased lipid droplets, smooth endoplasmic reticulum and mitochondriae in the treated group. The mRNA expression of IGF-I increased in the low dose group, and the protein expression of StAR decreased in the middle and high dose group.

CONCLUSIONLactating exposure to DEHP may interfere with the synthesis of testosterone of PLC in male pubs, the decrease of StAR and the damage of PLC may be involved in it.

Animals ; Body Weight ; Diethylhexyl Phthalate ; adverse effects ; Female ; Germ Cells ; drug effects ; Insulin-Like Growth Factor I ; metabolism ; Lactation ; Leydig Cells ; cytology ; drug effects ; Male ; Organ Size ; Phosphoproteins ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; drug effects ; Testis ; Testosterone ; blood

OBJECTIVETo investigate the etiologic value of diarrheagenic E. coil harboring genomic O island 28(OI-28) containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635), which were related to RTX (Repeat in toxin) toxin family isolated from children with diarrheal disease in Taiyuan.

METHODSIn the study, 257 fecal samples from children with diarrheal disease collected in Shanxi Children's Hospital. Diarrheagenic E. coli and enteropathogenic bacteria were isolated and identified by conventional bacterial culture and typing specific diarrheagenic E. coli (EPEC, EIEC, ETEC and EHEC) diagnostic serum, while diarrheagenic E. coli harboring genomic 01-28 containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635) were detected by PCR and DNA southern blot hybridization.

RESULTS206 strains (80.16%) of enteropathogenic bacteria were detected from 257 children with diarrhea disease, containing 149 strains (57.98%) of diarrheagenic E. coli and 57 strains(22.18%) of other entero-pathogenic bacteria. Among 3 strains (2.01%) of EPEC, 2 strains (1.34%) of ETEC, 2 strains (1.34%) EHEC were detected by typing specific serum, while all of the 142 strains (95.30%) isolated were suspected to be diarrheagenic E. coli. 21 strains (14.09%) of diarrheagenic E. coil harboring genomic O1-28 containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635) were detected by polymerase chain reaction and DNA southen blot hybridization, 8 strains (5.37%) of diarrheagenic E. coli containing only one genomic OI-28 virulence gene, 2 strains (1.34%) of diarrheagenic E. coli containing two genomic OI-28 virulence gene. 21 children with diarrhea diseases caused OI-28-harboring E. coli containing five important putative virulence genes were among 0 to 3 years old (80.95%). These children correlating with OI-28-harboring E. coli did not present special clinical symptoms or signs.

CONCLUSIONThe diarrheagenic E. coil harboring genomic OI-28 was one of the important etiology for children with diarrheal disease in summer season.

Child ; China ; Diarrhea ; microbiology ; Escherichia coli ; genetics ; pathogenicity ; Escherichia coli Infections ; complications ; Genes, Bacterial ; Humans ; Virulence