Objective To investigate the effect of CaMK Ⅱ expression on apoptosis of rat hepatocytes BRL-3A.Methods Rat BRL-3A cells were stable passage were cultured.The CaMK Ⅱ γ protein (LV-CaMK Ⅱ γ group) and CaMK Ⅱ γshRNA (shRNA group) lentiviral expression systems were constructed.The corresponding blank vectors (LV-NC group and shRNA-NC group) and normal saline (CON group) were perfused into the control groups.The expression levels of CaMK Ⅱ,Cyt C and MF proteins were detected by Western blotting,and the apoptosis rate of BRL-3A cells was measured by Tunel method.Results The protein expression of CaMK Ⅱ,Cyt C and AIF in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).The protein expression of CaMK Ⅱ,Cyt C and AIF in shRNA group was significantly lower than that in CON group (P＜ 0.05).There was no significant difference among CON group,LV-NC group and shRNA-NC group (P＞0.05).At the same time point,the apoptosis rate of hepatocytes in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).At the same time point,the apoptosis rate of hepatocytes in shRNA group was significantly higher than in CON group (P＜0.05).There was no significant difference in the apoptosis of hepatocytes among CON group,LV-NC group and shRNA-NC group (P＞0.05).Conclusion The specific CaMK Ⅱ signaling pathway can inhibit the apoptosis of BRL-3A cells,while the enhanced CaMK Ⅱ signaling pathway promotes the apoptosis of BRL-3A cells.

OBJECTIVETo analyse the relationship between the electrical activity changes of nucleus accumbens (NAc) shell and the drug-seeking behavior by recording NAc shell electrical activity in conditioned place preference (CPP) rats induced by morphine.

METHODSForty SD rats were randomly divided into operation-only control group and the morphine-induced CPP group after stereotaxic electrode was buried on rats NAc shell and the latter group was used to establish the morphine CPP model(n = 20). A CPP video system combining with the technique of electrical activity wireless telemetry was used in the study. The NAc electrical activity from each group of rats was recorded by wireless telemetry respectively, which included staying in black or white chamber of video box, shuttling between black-white chambers and between white-black chambers. The electrical activity differences were analyzed by the percentage of each wave.

RESULTSWhen the morphine-induced rats staying in black chamber, compared with the operation-only control group, the NAc shell electrical activity showed that the percentage of 0 - 10 Hz was increased(P < 0.05), meanwhile, those of 10 - 20 Hz and 30 - 40 Hz were reduced(P < 0.05, P < 0.01); when the morphine-induced rats staying in white chamber, the NAc shell electrical activity showed that the percentage of 0 - 10 Hz and 30 - 40 Hz were increased(P < 0.05 , P < 0.01) , that of 10 - 20 Hz was reduced(P < 0.05 , P < 0. 01); when the morphine-induced rats in black- white shuttling status, the NAc shell electrical activity showed that the percentage of 0 - 10 Hz was increased(P <0.05, P <0.01), that of 10- 30 Hz was reduced( P <0.05); and in the white-black shuttling status, the electrical activity showed that the percentage of 0 - 10 Hz was reduced(P <0.05), that of 10 - 30 Hz was increased(P < 0.05) ; the electrical activity was further compared between staying status and shuttling status in the morphine-induced CPP group. There was no significant difference of electrical activity between the rats in white-black shuttling status and staying in white chamber. However, when rats in black-white shuttling status, compared with staying in black chamber, the electrical activity showed that the percentage of 0 - 10 Hz and 40 - 50 Hz were increased(P < 0.05), meanwhile, those of 10 - 20 Hz and 30 - 40 Hz were reduced(P <0.05).

CONCLUSIONThe electrical activity changes of NAc shell in morphine-induced CPP rats were different from those of the operation-only control group, and these changes might be associated to the rat's drug-seeking behavior.

Animals ; Conditioning (Psychology) ; Drug-Seeking Behavior ; Morphine ; pharmacology ; Nucleus Accumbens ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Telemetry

Objective To approach a semi-quantitative diagnostic criteria of neonatal hypoxic-ischemic encephalopathy(HIE) by comprehensive scoring, including history, clinical manifestation and laboratory data,which can be used for early diagnosis,severity grading and the institution of therapy.Methods We analyzed history, clinical manifestation and some parameters of laboratory data,and established criteria for scoring and grading,and compared this grading with the grading determined by clinical data and follow-up,which brought forward a semi-quantitative diagnostic criteria, and applied this criteria to other 41 patients with HIE to test its reliability.Results The grading criteria were established as the following:if total score ≥20.0,the patient was graded as severe one;total score between 10.5-19.5, moderate one; between 5.5-10.0,mild one;≤5.0,non-HIE one. The scoring system was used in other 41 patients with HIE,the specificity and sensitivity were 100% and 96 .9%,97.2% and 100%,100% and 100%, respectively.Conclusions For severe HIE cases who are not permitted to receive compated tomography(CT) examination or no CT can be used, this scoring system can be used to diagnoze HIE and grade the severity. This method is simple and easy to perform.Both specificity and sensitivity are high for diagnosis and severity judgment of HIE.

Until the recent emergence/re-emergence of human-pathogenic viruses in ticks, tick-borne viruses have been neglected as causative agents of human disease (particularly in China). To gain insight into the diversity of tick-borne viruses in Xinjiang Uygur Autonomous Region (northwestern China), we conducted illumina deep sequencing-based screening for virus-derived small RNAs in field-collected Ixodes persulcatus ticks. We found 32, 631 unique virus-matched reads. In particular, 77 reads mapped to the tick-borne group within the genus of Flavivirus, and covered 3.8%-2.4% viral genomes. In addition, 32 unique reads were specific to the Siberian subtype of tick-borne encephalitis viruses (TBEV-Sib) which have never been reported in Chinese TBE loci. We confirmed the potential existence of TBEV-Sib by amplification (using reverse transcription-polymerase chain reaction) of genomic fragments from the envelope gene or 3' genomic terminus from the pools of examined ticks. Both sequences demonstrated high homology to TBEV-Sib strains attached geographically to southern Siberia with nucleotide identity of 97.2%-95.5% and aminoacid identity of 99.4%-98.3%, respectively. In conclusion, we report, for the first time, detection of TBEV-Sib in the natural TBE loci of China. These novel data may provide genetic information for further isolation and epidemiologic investigation of TBEV-Sib.
Animals ; Arachnid Vectors ; virology ; China ; Encephalitis Viruses, Tick-Borne ; classification ; genetics ; isolation & purification ; Encephalitis, Tick-Borne ; transmission ; virology ; Genome, Viral ; Humans ; Ixodes ; virology ; Molecular Sequence Data ; Phylogeny

Objective To investigate the trend of liver function changes in untreated adult patients with Graves’ disease in China, and to analyze the associated factors. Methods Patients with newly diagnosed as well as recurrent Graves’ disease from January 2006 to August 2014 were enrolled. They were over 18 years old and did not receive any treatments, Examination of liver function, thyroid function, and thyroid related antibodies as well as tests regarding virus hepatitis were performed. Results A total of 1 254 patients were enrolled. 33 patients with virus hepatitis were ruled out. Ultimately, 1 221 patients matched the criteria of our trial, with 347 males and 874 females [(39. 3 ± 9. 5) year old]. After inclusion, they were assigned to 2 groups according to their liver function results(605 in normal group and 616 in abnormal group). Compared to normal group, patients in the abnormal group were older [(40. 1 ± 9. 2 vs 38. 5 ± 8. 7) year old, P<0. 05] and with higher proportion of females(81. 8% vs 61. 2% , P<0. 05). Regarding the thyroid function and related antibody tests, some patients yielded results that were extremely high so as to exceed the upper limit of the normal range. These patients were more frequently seen in the group with abnormal liver function. The patients whose thyroid function parameters exceeded the upper limit had higher level of alanine aminotransferase[ALT,(37. 69 ± 7. 51 vs 31. 90 ± 5. 95) U/ L, P<0. 05], aspartate aminotransferase[AST, (31. 97 ± 5. 09 vs 27. 88 ± 3. 82) U/ L, P<0. 05], direct bilirubin[DBiL, (5. 58 ± 0. 77 vs 4. 54 ± 0. 71) μmol/ L, P<0. 05]than the group whose thyroid function on the detected range. In the patients with all results detected, patients in abnormal liver function group had higher level of triiodthyronine[T3 , (5. 42 ± 0. 29 vs 4. 94 ± 0. 33) nmol/ L, P<0. 05], thyroxin[T4 ,(217. 53 ±14. 32 vs 204. 22 ±13. 54) nmol/ L, P<0. 05], free triiodthyronine[FT3 ,(15. 88 ± 2. 86 vs 14. 48 ±4. 83) pmol/ L, P<0. 05], free thyroxin[FT4 ,(48. 91 ±8. 45 vs 42. 95 ±6. 14) pmol/ L, P<0. 05], thyroid peroxidase antibody[ TPOAb, (402. 75 ± 89. 99 vs 210. 70 ± 44. 63) IU/ ml, P < 0. 05] and thyrotrophin receptor antibody[TRAb,(14. 08 ± 5. 24 vs 9. 04 ± 2. 58) IU/ L, P<0. 05]. Multivariate logistic regression analysis revealed that patients’ age(OR=0. 98, 95% CI 0. 97-0. 99), gender(OR=0. 94, 95% CI 0. 91-0. 97), level of FT4 (OR=3. 08, 95% CI 2. 19-4. 32), TPOAb(OR = 0. 98, 95% CI 0. 97-0. 99), and TRAb(OR = 1. 07, 95% CI 1. 01-1. 12) were independent risk factors of their liver dysfunction. Conclusion Graves’ disease may lead to liver dysfunction, which is much more common and severe in elder and female patients, as well as patients who are suffering from hyperthyroidism and raised level of thyroid related antibodies.

ObjectiveTo assess the efficacy and safety of Xuebijing injection for the treatment of severe acute pancreatitis (SAP).Methods An extensive search of related literatures from the Cochrane Library, EMBASE, China Biology Medicine (CBM), CNKI, VIP and Wanfang data up to March 2014 was performed. Randomized controlled trials (RCTs) regarding Xuebijing injection for the treatment of SAP were collected regardless of languages. Jadad scale was taken for quality evaluation of the included studies by two researchers. The patients in control group were given conventional treatment, and those of the Xuebijing group were given Xuebijing injection on the top of conventional treatment. The Cochrane Collaboration RevMan 5.2 software was used for data analysis regarding the effect of Xuebijing injection on the mortality, incidence of complication, effective rate, the length of stay in hospital, and the safety of the drug in patients with SAP.Results A total of 15 published reports meeting the inclusion criteria were enrolled. The methodological quality of the trials was low. Meta analysis showed that the mortality in Xuebijing group was significantly lower [odds ratio (OR) = 0.37, 95% confidence interval (95%CI) =0.17 - 0.77,P = 0.008], and the incidence of complication was also significantly decreased (OR = 0.26, 95%CI =0.14 - 0.45,P< 0.000 01) as compared with those of control group. The effective rate in Xuebijing group was significantly higher than that of the control group [relative risk (RR) = 0.85, 95%CI = 0.80-0.91,P< 0.000 01]. The length of stay in hospital in Xuebijing group was significantly shorter than that of the control group [mean difference (MD) = -5.28, 95%CI = -6.69 to -3.86,P< 0.000 01]. Adverse reactions of Xuebijing injection were reported in 2 studies. The adverse reaction in one study was headache and nausea, which were relieved by adjusting the speed of intravenous infusion, and mild rash was reported in another case, and it disappeared after the withdrawal of Xuebijing. Conclusions The currently available evidence shows that Xuebijing injection may have some therapeutic effect on SAP. Because of the low methodological quality of the included trials, multi-center and high-quality RCTs with large sample sizes are needed to provide stronger evidence.

AIM:To explore the protective effects of exosome secreted by human umbilical mesenchymal stem cells on cardiac fibrosis in diabetic mouse model.METHODS:Male C57BL/6 mice at 6～8 weeks of age were divided in-to 3 groups randomly:control group ,diabetes mellitus(DM)group and DM+exosome group.To develop mouse DM mo-del,the mice were fed with high-fat diet for 5 weeks,followed by intraperitoneal injection of 45 mg/kg streptozocin once a week for 5 weeks.It was considered as a successful DM model that the blood glucose of the mice was ≥16.7 mmol/L.The mice in DM+exosome group were injected with exosome via tail vein.The mice in other 2 groups were injected with saline at the same volume.The heart function was evaluated by color Doppler echocardiography for small animals.The blood sam-ples were collected from abdominal aortas.The blood glucose and non-esterified fatty acids were measured by biochemical colorimetric assay.HE staining was performed to observe the structural changes of myocardial fibers ,and Masson staining was used to observe the cardiac fibrosis.RESULTS:The results of echocardiography showed that left ventricular end-dias-tolic dimension(LVIDd)and left ventricular end-systolic dimension(LVIDs)of diabetic mice were larger than those incontrol mice(P<0.05 and P<0.01,respectively).The ejection fraction(EF)and fractional shortening(FS)decreased in the diabetic mice(P<0.01).Exosome treatment significant decreased the LVIDs(P<0.01),but increased the EF and FS(P<0.01).The blood glucose and non-esterified fatty acids were significantly increased in the diabetic mice.The injection of the stem cell exosome significantly decreased the blood glucose and non -esterified fatty acids(P<0.01).HE staining observation showed that cardiomyocyte hypertrophy and fragmentation of cardiomyocyte in DM group were more se -rious than those in control group.Masson staining showed that the area of fibrosis in DM group was larger than that in con-trol group(P<0.01),but that in DM+exosome group was reduced(P<0.01).CONCLUSION:Exosome secreted by human umbilical mesenchymal stem cells protects the DM model mice from cardiac fibrosis.

China ; epidemiology ; Coronavirus Infections ; epidemiology ; prevention & control ; transmission ; Humans ; International Cooperation

Objective To observe whether the transplanted dermal multipotent stem cells(dMSCs)transfected by adenovirus vector of CXCR4(Adv-CXCR4)can distribute more frequently to the wound of rats with combined wound and irradiation injury.Methods dMSCs transfected by Adv-CXCR4(group A),or transfected by adenovirus vector of green fluorescent protein(group B),and non-transfected dMSCs were labeled with 3H-TdR and then transplanted into combine-injured rats.The amount of dMSCs in wound were determined by liquid scintillation,and wounds healing process was observed by measuring the remaining wound area.Results From the 5th day after transplantation,the amount of dMSCs in the wound of group A accounted for 1.95%-3.85% of the total transplanted dMSCs,significantly greater than those in group B and group C,which accounted for 1.07%-1.86% of the total transplanted dMSCs.The remaining wound area in group A was smaller than those in group B and group C from day 12 after injury,and the healing time of group A was 1.5 day ahead than group B and group C.Conclusions dMSCs transfected by Adv-CXCR4 distributes more frequently to the wound of combine-injured rats and could accelerate wound healing.

OBJECTIVETo investigate the effects of hydrogen sulfide preconditioning on myocardial ischemia reperfusion injury in rats.

METHODSSprague-Dawley male rats were divided into 4 groups with 10 in each group: in S group rats received sham operation; in IR group rats were given with NS (1.0 ml/kg iv) 24 h before ischemia; in H group rats were treated with NaHS (0.05 mg/kg iv) 24 h before ischemia; and in D group, NaHS-treated rats received 5-hydroxydecanoate (5-HD) 15 min before ischemia. Rats in IR group,H group and D group were subjected to ischemia by occlusion of coronary artery for 30 min followed by 2 h of reperfusion. At the end of the reperfusion,myocardial infarct size was measured. SAM-s was measured by Western blotting. Plasma SOD activity and MDA were determined at the end of reperfusion.

RESULTSThe infarct size was significantly lesser in H group (25.40 % ± 3.54%) than that in IR group (38.27% ±5.64%,P<0.05). The SAM-s protein expression in myocardium was significantly lower in H group than that in IR group. The plasma MDA content was significantly lower and SOD activity was higher in H group than those in IR group,but there was no difference between IR group and D group.

CONCLUSIONThe hydrogen sulfide preconditioning attenuates myocardial IR injury possibly through down-regulating SAM-s expression,reducing the production of oxygen free radicals and enhancing anti-oxidize effect in rats.

Animals ; Disease Models, Animal ; Hydrogen Sulfide ; pharmacology ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley