Objective To study the effect of CO2 pneumoperitoneum on migration of GFP-labeled living cells into the liver through blood route in rat model.Methods SD rats was inoculated intraportally with high-dose(5?106) GFP-labeled liver cells from C57BL/6 mice after cutting belly open.Pneumoperitoneum was established immediately after closing the abdominal wall.The rats were randomly divided into four groups(n=10 in each) to receive CO2 pneumoperitoneum at 5,10,or 15 mmHg,or no treatment other than cells inoculation(control).The pneumoperitoneum was maintained for 30 min.Afterwards,the rats were euthanized by cervical dislocation,and the liver of the rats was removed for fast frozen section biopsy.The expression of GFP-labeled living cells in rat livers was compared between the groups.Results No significant difference was detected in the positive expression of GFP-labeled cells between the groups(8 rats in the control group,9 in the 5 mm Hg group,9 in the 10 mm Hg group,and 10 in the 15 mm Hg group,?2=2.222,P=0.528).The mean number of GFP-positive cells in the four groups was 6.63?2.45(control),7.67?2.83(5 mm Hg),13.89?4.37(10 mm Hg),and 15.50?6.29(15 mm Hg).There was significant differences between the four groups(F=10.78,P=0.000).In addition,the numbers of GFP-positive cells in the high pressure groups(15 mm Hg and 10 mm Hg) were significantly higher than that in the low pressure group(5 mm Hg) and the control(P

Animals ; Colon ; physiology ; Female ; Indoles ; pharmacology ; Male ; Myoelectric Complex, Migrating ; Rats ; Rats, Wistar ; Viscera ; physiology

Asthenia ; physiopathology ; Cyanosis ; physiopathology ; Expert Testimony ; Fatigue ; physiopathology ; Heart Defects, Congenital ; physiopathology ; Humans ; Walking

The feature of HPM(high-power microwave) and its effect are briefly introduced.The destroying mechanism of HPM to different electronic medical equipment is described in detail.The reinforcement of HPM and effective measures are discussed.It is crucial to study the effects of high-power microwave radiation on military electronic equipment due to wide application of HPM in future military actions.

AIM: To better reveal the changing process of macula area after photodynamic therapy (PDT), the changes of tomographic imaging on macular fovea optical coherence tomography ( OCT ) scan at different follow- up time points after treating age-related macular degeneration ( AMD) by PDT were investigated.
METHODS: This cohort study included 30 eyes of 26 patients diagnosed as AMD. The dosage of verteporfin was calculated according to manufacturer's instruction. All participants got FFA, ICG, OCT scan and best corrected visual acuity before and followed up for 2wk, 1, 2, 3mo after PDT. The standard five lines combined with 6 lines OCT scan covered key parts of lesion and the scan locations before and after were kept the same. The bilaminar foveal thickness (BFT), outer high reflectivity band thickness ( OHRBT ) , and the total area of intraretinal fluid ( IRF ) and subretinal fluid ( SRF ) were measured at different time points. The relationship between the changes and follow up time was analyzed. Statistical analysis of the data was performed using SPSS for windows version 13. 0.
RESULTS: There months after treatment, an improvement of 2 lines or better on the Snellen chart was achieved on 22 eyes, visual acuity of 3 eyes without significant change, 3 eyes got decreased, 2 cases lost follow up. The average macular thickness of improved 22 eyes before treatment was 722. 5±55. 6μm, 2wk after treatment, 708. 3±45. 3μm, 1mo, 584. 4±49. 3μm, 2mo, 430. 7±50. 2μm and 180. 6 ±36. 3μm at 3mo. The OHRBT before and after treatment respectively were 302. 3 ±50-2μm, 277. 5±42. 3μm, 202. 7±40. 1μm, 180. 6±35. 7μm, 100. 8±22. 9μm. The total area of both IRF and SRF was estimated as 0. 34±0. 12mm2 , 0. 25±0. 07mm2 , 0. 10±0-05mm2 , 0. 08±0. 04mm2 , 0. 05±0. 01mm2 .
CONCLUSION:SRF and retinal edema of patients with AMD were absorbed 1mo after PDT, and the BFT, OHRBT were significantly tend to be thin.

Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.

Objective To study skeletal muscle atrophy and the change of autophagy in skeletal muscle of patients with chronic kidney disease.Methods Mean muscle cross sectional area,mRNA and protein expression of autophagy markers Bcl-2-adenovirus E1B interacting protein 3 (LC3B),Bcl-2-adenovirus E1B interacting protein 3 (Bnip3),Beclin-1 were measured in rectus abdominis biopsies obtained from 22 consecutive patients with stage 5 CKD scheduled for peritoneal dialysis from 4 hospitals in Shanghai.Control biopsies were obtained from another 8 healthy subjects during elective surgery for adenomyosis and 6 subjects during elective surgery for abdominal wall hernias.Rectus abdominis muscles were obtained at the beginning of surgery.HE staining was performed and mean cross sectional area (CSA) was calculated.Electron microscopy was used to confirm the changes of autophagy.mRNA levels of LC3B,Beclin-1,Bnip3 were evaluated by RT-PCR and protein levels of those parameters were evaluated by Western blotting.Results Compared with control group,mean CSA of muscle fibers was decreased and the transcript levels of LC3B,Beclin-1,Bnip3 were up-regulated in CKD group.Similarly,protein levels of LC3BⅠ,LC3B Ⅱ,Beclin-1 and Bnip3 were increased in CKD group.Additionally,activation of autophagy was confirmed by the appearance of autophagosomes by electron microscopy.Conclusion Chronic kidney disease may cause skeletal muscle atrophy and lead to activation of autophagy,which may contribute to muscle atrophy.

Objective:To investigate the chemokine receptors expression on regulatory T cells induced by different antigens and chemotaxis of T cells conducted by CCL20 and CCL22.Methods:BALB/c mice were divided into different groups and inoculated with skin-antigen derived from C57BL/6 mice or BCG vaccine respectively.The changes of CCR4 and CCR6 expression on CD4+CD25-T cells and CD4+CD25+CD127-T cells were detected at 1st,2nd,3rd and 4th week using flow cytometer.The chemotactic effects of CCL20 and CCL22 on CD4+CD25-T cells and CD4+CD25+CD127-T cells subsets were assayed by chemotaxis assay.Results: ①In skin-antigen group,the average fluorescence intensity ( MFI) of CCR4 on CD4+CD25-T cells at 4 week was significantly stronger than that at 3 week (P<0.05).There were no significant changes of CCR4 expression in BCG group.②The MFI of CCR6 on CD4+CD25-T cells was strongest at 2 week in skin-antigen group (P<0.05) while in other groups at 4th week (P<0.05).Besides,the expression of CCR6 on CD4+CD25+CD127-T cells was stronger during the first two weeks than the later two weeks ( P<0.05) in skin-antigen group, while in BCG group,the MFI was stronger at 2nd and 4th week than 1st week (P<0.05).③The chemotactic index of CD4+CD25+CD127-T cells was highest at 4th week in BCG group (P<0.05) in CCL20 induced chemotaxis,while in other groups were higher at 2nd and 4th week(P<0.05).In CCL22 induced chemotaxis ,there were no significantly differences of chemotactic index of CD4+CD25+CD127-T cells between skin-antigen group and BCG group.Conclusion:①The expression of chemokine receptor on the surface of Treg was associated with antigenic properties.②CCL22 had a notable chemotactic effect on Treg at the early stage of post-induction, while CCL20 did that at the late stage of post-induction.

Objective To establish a reverse transcription nested polymerase chain reaction ( RT-nPCR ) assay for detection of tree shrews orthoreovirus (TRV).Methods Three strains of TRV were respectively isolated from fresh feces of three tree shrews that came from the same field at different times .We designed and synthesized two pairs of MRV L1 gene nested primers and established the system of RT-nPCR.The TRV RNA was extracted and reversely transcribed to cDNA as a template for nested-PCR amplification.The developed RT-nPCR was optimized.The specificity and sensitivity were tested.Finally, the RT-nPCR was used to detect TRV in 25 tree shrew samples.Results Taking the genomic RNA of TRV as template, the RT-nPCR was able to amplify a specific fragment band targeting the L 1 gene, while there were no target bands in the normal cell control , ( Wa strain rotavirus , hepatitis A virus , and herpes simplex virus ) .The RNA of TRV was diluted by 1:10 to 1:109 .Each dilution sample was analyzed by the RT-nPCR.The minimum detectable concentration of RNA was 0.01 pg/μL.The results of RT-nPCR detection showed that 4 of the 15 tree shrews were TRV-positive in the survival group , and 10 of 10 tree shrews were TRV-positive in the death group . Conclusions The RT-nRCR assay established in this study is accurate , specific and sensitive .Therefore, it can be used for routine detection of TRV in quality assurance testing .

Objective To identity whether there is muscle atrophy phenomenon in end-stage kidney disease patients and to detect the level of transcription factor Foxo1 and the activity of ubiquitin-proteasome system.Methods Twenty-two patients in chronic kidney disease (CKD) stage 5 were selected and their mean muscle cross sectional area was measured.mRNA and protein levels of Foxo1,Atrogin-1,MuRF1 in rectus abdominis biopsies obtained from consecutive patients were detected.Control biopsies were obtained from 8 healthy subjects during elective surgery for abdominal wall hernias and 6 subjects during elective surgery for adenomyosis.Results Compared with the control group,cross sectional area of muscle fibers decreased and the transcription and protein levels of Foxo1,Atrogin-1,MuRF1 were upregulated in CKD group(P＜0.05).Protein level of p-Foxo1 decreased in CKD group(P＜0.05).Conclusion There exist muscle atrophy phenomenon in CKD patients,which may associate with the upregulation of Foxo1 and activation of ubiquitin-proteasome system.