Objective To evaluate the effect of follicular fluid(FF)exosomal miRNAs on follicular dysplasia in patients with polycystic ovary syndrome(PCOS)mediated by glycolysis pathway of granulosa cells(GCs),and to explore the mechanism. Methods Three PCOS infertile patients and three non-PCOS infertile patients were recruited. The baseline hormone levels of the two groups were measured before ovulation induction. The bilateral FF was obtained by puncture after short-acting and long-term ovulation induction,and the exosomes were collected by ultracentrifugation and identified by transmission electron microscopy. The total exosomal RNA was extracted by Trizol method to construct the library,which was compared to the reference genome GRCh38 for statistical analysis after miRNA sequencing and quality control processing. Clustering Profiler R package was used to implement GO annotation analysis and KEGG pathway analysis of the differentially expressed genes(DEGs),and Omnipath software for miRNAs interaction analysis. A total of 16 miRNA were randomly selected and detected by qPCR to verify the accuracy of the miRNA sequencing results. Results Compared with the non-PCOS group,luteinizing hormone(LH),anti-Muerian hormone(AMH),testosterone and antral follicle counts in PCOS group increased significantly(t = 2. 479 ～ 9. 163,each P < 0. 05). The exosomes of FF in both groups showed the cup-shaped vesicles with clear edge and light staining in the center,with the diameters of 100 — 150 nm and intact structure,and the concentration was about 8 × 1010particles/mL. A total of 928 miRNAs were detected by miRNA sequencing. Compared with the non-PCOS group,59 differentially expressed miRNA(DEmiRNA)were screened out in exosomes of POCS group,of which 31 were up-regulated and 28 were down-regulated. The differential trend of gene expression detected by qPCR was highly similar to that of miRNA sequencing. In FF exosomes of PCOS patients,the glycolysis efficiency and apoptosis of GCs were significantly changed by miRNA regulating mRNA. PKM,PFKL and HK2 were the key target genes for miRNA to regulate GCs glycolysis,and SLC2A1 was the key target gene for miRNA to regulate GCs apoptosis. Conclusion The miRNAs in FF exosomes of PCOS patients can weaken the glycolysis of GCs while accelerate the apoptosis,thus reducing the production of ATP and lactic acid,resulting in follicular dysplasia.

Accumulation and infiltration of polymorphonuclear leukocytes are pivotal factors in accelerating cell apoptosis and death in ischemia/reperfusion(I/R) injury, however, adhesion molecules play an important role in its rolling，latching and infiltrating. The goal of the review is to explore the effect and significance of intercellular adhesion molecule-1(ICAM-1) in ischemia/reperfusion.

Objective To investigate the effect of autologous transplantation of circulating endothelial progenitor cells (EPCs) on oleic acid-induced acute lung injury (ALI) in rabbits. Methods Thirty New Zealand long ear rabbits weighing 1.8-2.0 kg were randomly divided into 2 groups ( n = 15 each): normal saline group (group NS) and EPC group. ALI was induced by iv oleic acid 80 mg/kg. EPC (106/200 μl) or equal volume of normal saline (NS) was administered iv at 4 h after iv oleic acid injection. Arterial blood samples were obtained before (T0) and at 4, 8, 12, 24 and 48 h (T1-5) after oleic acid injection for blood gas analysis and determination of serum vascular endothelial growth factor (VEGF) concentration. The animals were then scrificed (at T5 ) and the lungs were removed for microscopic examination and determination of W/D lung weight ratio, Pulmonary infiltration of PMN and non-PMN was counted and hyaline membrane formation and hemorrhage were examined. Results PaO2/FaO2 ratio and serum VEGF concentration were significantly higher in group EPC than in group NS. Infiltration of leukocytes in the lung was significantly reduced by EPC transplantation. EPC also decreased lung water content, hyaline membrane formation and hemorrhage in the lungs. Conclusion Autologous transplantation of circulating EPC can ameliorate oleic acid-induced acute lung injury in rabbits.

Because of the disunity use of the outcome measures in traditional Chinese medicinal clinical trials, we explain the application of outcome measures in clinical trials on insomnia, provide the international criteria and guideline on outcome reporting. According to analysis of the concrete cases, we in order to improve the misuse of outcome measures of insomnia, to make the selection and reporting of outcome measures follow with the international practices and, to accelerate the development of traditional Chinese medicine.

Objective To study the effect of monosialoteterahexosyl ganglioside (GM1) on neurobehavioral development in premature infants with white matter damage. Methods A total of 636premature infants who were hospitalized in NICU of two hospitals from Jan 2005 to May 2009 received routine bedside cranial sonography detection before 1 week-aged. Forty premature infants were diagnosed as being premature white matter damage and divided into the treatment group (20 cases ) and the control group (20 cases) randomly. The cases in the treatment group accepted GM1 20 mg additional to 5% glucose solutionthe iv drip, one time per day,for a cycle of 14 d. 1 -3 cycles were given in accordance with patient's condition. Other treatments were same to the control group. All cases were evaluated by neonatal behavioral and neurological assessment (NBNA) at the rectified age of 40 gestational weeks and by Children's Developmental Center of China (CDCC) test at 3 months-aged and 12 months-aged. Results The NBNA scores of the treatment group (38.10±0.91) were significantly higher than the control group (36.10±1.59) at the rectified age of 40 gestational weeks (P＜0.01). The indexes of mental development(MDI) and psychomotor performance development (PDI) by the CDCC tests in the treating group (3 months-aged MDI:91.66±6.38;PDI:87.11±5.57; 12 months-aged MDI:104.10±6.45; PDI:100.46±3.87) were significantly higher than those in the control group (3 months-aged MDI:81.07±0.72; PDI:81.90±6.70; 12 months-aged MDI:98.45±8.57; PDI:95.91±6.59) at 3 months-aged and 12 months-aged ( P ＜ 0. 05 ). Conclusion GM1 can accelerate the neurobehavioral development in premature infants with white matter damage.

Objective To clone and express the cDNA encoding Schistosoma japonicum tropomyosin. Methods The cDNA was amplified by reverse transcription polymerase chain reaction (RT PCR). The PCR products were ligated with pGEM T vectors and then for transformations. After characterization of white clones by agarose gel electrophoresis, endonucleases digestion and PCR, some recombinant plasmids with inserts were used for sequencing. Then the gene was subcloned into prokaryotic expression vector pQE30 and expression was induced by IPTG. Results The PCR products was 823 bp judged by agarose gel electrophoresis and sequencing. A cDNA encoding S japonicum tropomyosin, except for 14 amino acids at the amino terminus and 2 at the carboxyl terminus, has been constructed and cloned successfully. The colony, designated pGSjcTM12, was sequenced and shown to be 91 1% identical at the nuclei acid level and 98.1% identical in deduced amino acid sequence to that of S mansoni tropomyosin. The gene was subcloned into pQE30 and an expressed protein of about 32 kDa was obtained.Conclusion The cloning and expression of the gene encoding S japonicum tropomyosin had been successfully made.

Acupuncture Therapy ; Adult ; Aged ; Angina, Unstable ; drug therapy ; physiopathology ; therapy ; Cardiovascular Agents ; administration & dosage ; Combined Modality Therapy ; Electrocardiography ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged

Objective A retrospective analysis was conducted to review the distribution and antibiotic resistance in the Staphy-lococcus aureus strains isolated from sputum and provide evidence for effective prevention and control of infections.Methods The 276 strains of S .aureus isolated during the period from January to September of 2009 were included in this analysis.The isolates were identified and tested for susceptibility using VITEK 2 Compact Automatic Culture and Identification System. Results The prevalence of methicillin-resistant S .aureus (MRSA)was 79.0% (218/276).The S .aureus strains were mainly isolated from Department of Neurology (27.5%,76/276),ICU (25.4%,70/276),Neurosurgery (18.1 %,50/276),and Re-spiratory Medicine (9.1 %,25/276).Vancomycin,teicoplanin and quinupristin-dalfopristin were the most active agents against these S .aureus .The resistant strains were less than 0.5%.MRSA showed lower resistance rate to trimethoprim-sulfamethox-azole than MSSA.MRSA strains were more resistant than MSSA to most other antibacterial agents.Conclusions The MRSA i-solates from sputum are highly prevalent and resistant to antibiotics.Antimicrobial agents should be selected appropriately ac-cording to the results of susceptibility testing.

Objective Pancreatic neuroendocrine tumor( PNET) is a rare kind of tumor with slow growth for which surgery is the main treatment.The histological grading of PNET was closely related to the choice of operation mode and the comprehensive treat-ment after operation.Imaging examinations such as enhanced CT and MRI are of great value in the preoperative diagnosis of PNET. The aim of this study was to investigate the characteristic features of CT and MRI in the PNET. Methods Preoperative imaging data of 80 patients with PNET confirmed by pathology were reviewed, including CT and MRI, and the image features of each PNET were an-alyzed and the statistical analysis was performed. Results There were significant differences in the lesion morphology between the G1 and G2 levels of PNET.The majority of forms of the G1 stage PNET lesions presented round-like regular shape, but the G2 stage did not show regular shape.CT examination showed that regular morphological lesions accounted for 91.30%(21/23) in the G1 level, and 52.00%(13/25)in the G2 level,χ2 =8.857,P=0.003.MRI examination showed that regular morphological lesions accounted for 88.48%(22/27)in the G1 level, and 55%(11/20) in the G2 level, χ2 =3.862,P=0.050.The difference of scan density in PNET lesions was statistically significant.Low density lesions accounted for 73.91%(17/23)in the G1 level, and 32%(8/25)in the G2 level.Equidensity lesions accounted for 26.09%(6/23)in the G1 level, and 68.00%(17/25)in the G2 level,χ2 =8.842,P=0.004.The difference of bile pancreatic duct expansion in the G1 and G2 level lesions was statistically significant by MRI examination. Lesions with expanded pancreatic duct accounted for 7.41%(2/27)in the G1 level,and 40%(8/20) in the G2 level, χ2 =7.287, P =0.007.There was significant difference in the peak time after en-hanced scanning between the G1 and G2 levels.Lesion enhancement peak at the arterial phase by CT examination of accounted for 21.74%(5/23) in the G1 level,and 64%(16/25) in the G2 level,χ2 =8.694,P=0.003.Lesion enhancement peak at the arterial phase by MRI examination accounted for 29.63% (8/27) in the G1 level, and 70% (14/20) in the G2 level, χ2 =7.521,P=0.006. Conclusion The lesion morphology, density of CT plain scan, CT or MRI enhancement peak time, and expansion of the bile duct and pancreatic duct can provide reliable information to distinguish the G1 and G2 stage in PNET.

ObjectiveTo construct and identify a DNA vaccine of Schistosoma japonicum (Chinese strain) thioredoxin. MethodsAccording to a cDNA sequence of S.japonicum (Philippine strain) thioredoxin, a couple of primers was designed with the BamH I restriction endonuclease site introduced in forward primer with ATG as start condon and EcoR I in reverse primer with TCA as termination codon. The gene encoding S.japonicum (Chinese strain) thioredoxin (SjcTrx) was amplified by RT-PCR using total RNA of schistosome as the template. The PCR products and the pcDNA3 plasmids were digested by restriction endonucleases BamH I and EcoR I. The target DNA fragment were purified and cloned properly into the eukaryotic expression vector of pcDNA3. The recombinant plasmids were then transformed into competent E.coli JM109 and identified by endonucleases digestion, PCR, agarose gel electrophoresis and sequencing. ResultsThe RT-PCR product was around 334 bp judged by agarose gel electrophoresis. The same fragments were obtained by restriction enzyme digestion from the recombinant plasmid and PCR with the plasmid DNA as a template. The recombinant plasmid, designated pcDNA3-SjcTrx, was sequenced and shown to be 97% and 43% identical in deduced amino acid sequences to that of S.japonicum (Philippine strain) and S.mansoni thioredoxin, respectively. ConclusionThe S.japonicum thioredoxin DNA vaccine had been constructed successfully, and further studies will be made in different animal models for its immunogenicity.