Objective:To explore the effect of cholesterol on the expression of genes for matrix synthesis and degradation of human meniscal fibrochondrocytes and its mechanism.Methods:Meniscal tissue was taken from patients undergoing arthroscopic surgery to extract fibrochondrocytes. The cells were divided into a control group in which the normal cells were not processed, a positive control group in which interleukin-1 β was used to create a degeneration model, and 2 treatment groups which were subjected to treatment with 15 and 30 μg/mL cholesterol respectively. Safranin O staining, β-galactosidase staining and enzymic kits were used to detect the morphology and total cholesterol (TCH) content of meniscal fibrochondrocytes in the 4 groups. Immunofluorescence and western blot were used to detect the protein expression of type Ⅰcollagen precursor α1 (COL1A1) and type Ⅱ collagen precursor α1 (COL2A1). RT-qPCR was used to detect the mRNA expression of COL1A1, COL2A1, matrix metalloproteinase (MMP) 3, MMP9, MMP13, and genes related to cholesterol efflux pathways [like liver X receptor α (LXR α), ATP binding cassette transporter A1 (ABCA1) and ABCG1]. Results:There was no significant difference between the control and the positive control groups in the TCH content in human meniscal fibrochondrocytes ( P>0.05). The treatments with 15 and 30 μg/mL cholesterol resulted in significantly increased TCH contents in human meniscal fibrochondrocytes in the treatment groups ( P<0.05). Compared with the control group, the mRNA expression of LXR α, ABCA1 and ABCG1 was significantly decreased in the treatment groups ( P<0.05), and the meniscal fibrochondrocytes in the positive group and the treatment groups presented with a lower density, chaotic distribution and obvious signs of degradation. Compared with the control groups, the mRNA expression of matrix synthesis genes (COL1A1 and COL2A1) in the meniscal fibrochondrocytes was significantly inhibited while the mRNA expression of matrix degradation metalloenzymes (MMP3, MMP9 and MMP13) was significantly promoted ( P<0.05). Conclusion:Cholesterol may inhibit the cholesterol efflux pathways of meniscal fibrochondrocytes, and thus cause accumulation of cholesterol in the meniscal fibrochondrocytes, eventually leading to degeneration of meniscus.

Objective To investigate the therapeutic method of metabolic acidosis in long-term sur- vival patients undergoing simultaneous pancreas and kidney transplantation.Methods A 45-year-old fe- male patient,who had undergone simultaneous pancreas and kidney transplantation(due to diabetic ne- phropathy and uremia)with bladder drainage 2 years before,developed severe metabolic acidosis,and thus underwent surgical conversion from bladder to ileal drainage.The procedure was as follows.The stoma of duo- denocystostomy was isolated and resected.The site of cystostomy was closed in two layers.The graft duode- num was then anastomosed to a loop of the recipient's ileum,which was proximal 40 cm from the ileocecum in a side-to-side manner.Results The metabolic acidosis resolved postoperatively.The patient received conventional immunosuppressants.The hospital stay was 30d.Follow-up of 4 years showed normal pancreas and kidney functions.Conclusions Conversion from bladder to ileal drainage is safe and effective for metabolic acidosis related to the exocrine secretions of bladder drained pancreas graft in simultaneous pancre- as and kidney transplant recipients.

[Objective]This study was designed to investigate the genetic evolution of the neuraminidase(NA)gene of seasonal A/H1N1 and 2009 novel A/H1N1 inflilenza virus,and discuss the genetic variation of influenza A virus.[Methods]The virus strains were separately isolated from the clinical samples collected in 2006 and 2009,and then identified as seasonal A/H1N1 and novel A/H1N1.The full length of the NA gene of these strains was amplified by RT-PCR.Then the genetic evolution and mutations of important functional sites were analyzed.[Results]The homology of NA gene between the 2009 novel A/H1N1 isolates and 2006 seasonal A/H1N1 isolates was low(77.9%～78.8%),so was the homology of NA gene between the 2009 novel A/H1N1 isolates and representative strains of different periods and 1979-2001 WHO recommended vaccine strains(78.1%～79.3%).But compared with the WHO recommended vaccine strains of 2009 novel A/H1N1,the homology reached more than 99%.The genetic evolution analysis revealed that NA gene of 2009 novel A/H1N1 had the closest genetic relationship with the swine influenza A virus(A/swine/Belgium/1/1983)from Eurasian Iineage,and some of the antigenic sites and neuraminidase active sites of NA gene of seasonal A/H1N1 were mutated after 2005.[Conclusion]The NA gene of 2009 novel A/H1N1 may originate from Eurasian Iineage of swine influenza virus.The variation of NA gene of seasonal A/H1N1 has occurred in a certain degree.Hence,it is very necessary to continuously monitor the variant of influenza A virus.

Objective To investigate the immunization coverage of oral live attenuated rotavirus vaccines and its influencing factors among children under 5 years old in Jinshan District of Shanghai in order to provide references for optimizing vaccination management in this area. Methods A cross-sectional study was conducted in 6 community vaccination clinics randomly selected from 12 vaccination clinics in Jinshan District. In each selected clinic,145 children under 5 years old were continuously recruited. Information on immunization status,demographics of children,family socioeconomic status and parents′knowledge of vacci-nation were collected through questionnaire interview. Logistic regression was used to analyze factors influen-cing immunization status by using SPSS22.0. Results Among the 863 participating children, the immuni-zation coverage rate of oral live attenuated rotavirus vaccines was 61.07%. Multivariate analysis revealed that children having an urban registration status (OR=2.21),better family income (OR=1.47) and par-ents with high recognition of rotavirus diarrhea (OR=8.56) and stronger intention to following "doctor′s suggestions" (OR=1.96) were more likely to accept oral live attenuated rotavirus vaccines, while those from families with a medical professional were less likely to be vaccinated (OR=0.57). Conclusion The immunization coverage rate of oral live attenuated rotavirus vaccines among children in Jinshan District were influenced by various factors such as children′s household registration status, household income, parents′knowledge regarding vaccination and doctors′suggestions. Health education on immunization should be strengthened and tailor-made to improve the coverage of oral rotavirus vaccination.

OBJECTIVETo explore the synthetical index for diagnosing schistosomiasis with ultrasound and to assess the prevalence rate with the index.

METHODSUltrasound indexes of schistosomiasis Japonicum were analyzed by principal component analysis, and the synthetical indexes were assessed by ROC curve.

RESULTSAmong the abnormal rates of the 6 indexes, the lowest was 1.6% comparing with the highest of 59.5%. Significant difference was noficed among the abnormal rates (chi(2) = 631.1, P < 0.01). The individual correlation of the six indexes to each other as will as with age distribution was significant (P < 0.05). The three principal components reflected the degree of pathological changes on liver and spleen. The first principal component was the factor reflecting the degree of liver pathological changes, and the second and third principal components reflected the degree of pathological changes on spleen. The synthetical index D(1) = 0.047X(1) + 0.428X(2) + 1.247X(3) + 0.095X(4) + 0.002X(5) + 0.213X(6) - 12.837 was found by adding the three weight principal components, and it's area under the ROC curve was 0.957. When -1.70 was taken as the critical value, the abnormal rate of population was 66.3%, close to the resident's actual prevalence rate 66.9%.

CONCLUSIONUltrasonography was considered as a method which could rapidly assessing the resident's prevalence rate in the endemic areas of schisitosomiasis Japonicum, and could also provide powerful information for development of strategy on chemotherapy.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Principal Component Analysis ; ROC Curve ; Schistosomiasis japonica ; diagnostic imaging ; epidemiology ; Ultrasonography

OBJECTIVETo study the expression changes of neuroglobin in rats with the model of diffuse traumatic brain injury and explore the relationship between the neuroglobin and neuron apoptosis in traumatic brain injury.

METHODSThe diffuse traumatic brain injury of rats was induced by the Marmarou's 'weight-drop' device. And the immunohistochemical technique was used to detect the expression changes of neuroglobin and neuron apoptosis in rat brain at different time points post-injury.

RESULTSThe expression of neuroglobin increased twice and reached peaks at 2 hours and 72 hours post-injury respectively. And the increased expression of neuroglobin from 30 minutes to 1 hour post-injury and from 48 hours to 72 hours post-injury accompanied with the decreased expression ratio of Bax to Bcl-2.

CONCLUSIONThe increased expression of neuroglobin in traumatic brain injury informed us that neuroglobin had anti-apoptosis action in post-injury neuron. It could protect the neuron from traumatic stress and secondary ischemia and hypoxia insults during ultra-early and acute stages.

Animals ; Apoptosis ; physiology ; Brain ; metabolism ; pathology ; Brain Injuries ; metabolism ; pathology ; Globins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Neurons ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVEThis study is to verify the use of rich BHI medium to substitute synthetic media for gene regulation studies in Yersinia pestis.

METHODSThe transcriptional regulation of rovA by PhoP or via temperature upshift, and that of pla by CRP were investigated when Y. pestis was cultured in BHI. After cultivation under 26 °C, and with temperature shifting from 26 to 37 °C, the wild-type (WT) strain or its phoP or crp null mutant (ΔphoP or Δcrp, respectively) was subject to RNA isolation, and then the promoter activity of rovA or pla in the above strains was detected by the primer extension assay. The rovA promoter-proximal region was cloned into the pRW50 containing a promoterless lacZ gene. The recombinant LacZ reporter plasmid was transformed into WT and ΔphoP to measure the promoter activity of rovA in these two strains with the β-Galactosidase enzyme assay system.

RESULTSWhen Y. pestis was cultured in BHI, the transcription of rovA was inhibited by PhoP and upon temperature upshift while that of pla was stimulated by CRP.

CONCLUSIONThe rich BHI medium without the need for modification to be introduced into the relevant stimulating conditions (which are essential to triggering relevant gene regulatory cascades), can be used in lieu of synthetic TMH media to cultivate Y. pestis for gene regulation studies.

Bacterial Proteins ; genetics ; metabolism ; Bacteriological Techniques ; Culture Media ; pharmacology ; Gene Expression Regulation, Bacterial ; drug effects ; physiology ; Yersinia pestis ; metabolism ; physiology

OBJECTIVETo understand the genetic structures and variations of the superintegron (SI) in Vibrio cholerae isolated in the seventh cholera pandemic.

METHODSPolymerase chain reaction scanning and fragment sequencing were used. Sixty toxigenic V. cholerae O1 El Tor strains isolated between 1961 and 2008 were analyzed.

RESULTSSome variations were found, including insertions, replacements, and deletions. Most of the deletions were probably the result of recombination between V. cholerae repeat sequences. The majority of the variations clustered together. The SIs of the strains isolated in the 1960s and 1970s showed more diversity, whereas SI cassette variations in strains isolated in the 1990s and after were lower, with ∼24 kb signature sequence deletion. This indicates the predominant SI in the host during the epidemic in the 1990s and after. The insertion cassettes suggested the mobilization from the SIs of other V. cholerae serogroups and Vibrio mimicus.

CONCLUSIONThe study revealed that structural variations of SIs were obvious in the strains isolated in epidemics in different decades, whereas the divergence was based on syntenic structure of SIs in these El Tor strains. Also, the continuing cassette flows in the SIs of the host strains during the seventh cholera pandemics were displayed.

Cholera ; epidemiology ; microbiology ; Chromosomes, Bacterial ; genetics ; ultrastructure ; Cluster Analysis ; DNA, Bacterial ; genetics ; Electrophoresis, Gel, Pulsed-Field ; Gene Deletion ; Gene Flow ; Genetic Variation ; Humans ; Integrons ; genetics ; Mutagenesis, Insertional ; Open Reading Frames ; genetics ; Polymerase Chain Reaction ; Tandem Repeat Sequences ; Vibrio cholerae O1 ; classification ; genetics ; isolation & purification ; ultrastructure

Objective To survey the prevalence rates of adult carotid atherosclerosis in water-sourceoriginated high iodine area in Shandong province in 2008 and discuss the relationship between water-sourceoriginated high iodine and carotid atherosclerosis occurrence. Methods Yuncheng county was chosen as observation area which included Dachenlou, Weigai, Liuyi and Ershilipu villages. Juye county was chosen as control that included Zhangxi, Bihainan, Xiaguan and Qiji villages. Two hundred and ninety-nine participants aged over 40 from water-source-originated high iodine areas and 323 residents aged over 40 from normal iodine areas were investigated.Portable-type B mode color Doppler was performed to examine the carotid artery of all participants. The adult carotid atherosclerosis was diagnosed and graded through the ultrasonograms. Results The prevalence rates of adult carotid atherosclerosis in the 4 water-source-originated high iodine villages of Dachenlou, Weigai, Liuyi and Ershilipu were 47.1% (33/70) ,62.2% (51/82) ,67.5% (52/77) and 58.6% (41/70), respectively and the prevalence rates of adult carotid atherosclerosis in the 4 normal iodine villages of Zhangxi, Bihainan, Xiaguan and Qiji were 40.7%(35/86),40.8% (31/76),38.2% (34/89) and 37.5% (27/72), respectively. The prevalence rates of adult carotid atherosclerosis standardized by age in the 4 high iodine villages and the 4 normal iodine villages were 45.81% ,58.18% ,61.63%,55.34% and 34.66%,36.25%,43.01% ,41.30%, respectively. The prevalence rates were higher in high iodine villages than that in control villages(T = 26, P ＜ 0.05 ). There were 136 people scored "0", 120 people scored "1 -3",43 people scored "4 - 7", and total 299 people. There were 201 people scored "0", 87 people scored "1 - 3", 35 people scored "4 - 7", and total 323 people. The expected prevalence rates and severity of adult carotid atherosclerosis was significantly different between high iodine area and normal iodine area (x2 = 17.54,18.42, all P ＜ 0.01).Conclusion The prevalence rate and severity of adult atherosclerosis in water-source-originatod high iodine area is higher than that in normal iodine area.

Objective To explore the efficacy and safety of autologous peripheral blood stem cell transplantation (APBSCT) in the treatment of systemic lupus erythematosus.Methods Nine patients with systemic lupus erythematosus were enrolled in this study.Patients were given cyclophosphamide and granu- locyte colony-stimulating factor(G-CSF)as the mobilization regimen.Urine was alkalinized and hydrolyzed to protect the function of the heart,liver and kidney of the patients.A CS3000 Plus blood cell separator was used to collect peripheral blood stem cells,which were preserved in liquid nitrogen.Two to five days before the administration of the stem cells,the patients were pretreated with intravenous injection of cyclophos- phamide (50 mg?kg~(-1)?day~1) for 4 consecutive days and antithymocyte globulin (ATG,2.5 mg?kg~(-1)?day~1) for 3 consecutive days.Granulocytes were recoverd by G-CSF stimulation.Then,the peripheral blood stem cells were reinfused.Therapeutic effect was evaluated by assessment of alteration of clinical manifestation (skin erythema),levels of proteinuria and antoantibodies,hematopoietic reconstitution and occurrence of transplantation related complications.Results After transplantation,all patients had been successfully en- grafted.The time for peripheral leucocyte count to reach 1.0?10~9/L was 7～15d;the time for platelets to reach 20?10~9/L was 0～21 d.The skin erythema resolved in all patients;proteinuria decreased to normal level and the autoantibodies became negative in most of the patients.Serum sickness-like response occurred in all patients,renal and heart failure in 1 patient,hemorrhagic cystitis in 3 patients,psychiatric disorders in 1 patient,candidal infection in 1 patient.Conclusion One-year follow up suggests that autologous stem cell transplantation is markedly effective and relatively safe for systemic lupus erythematosus.However,the duration of remission remains to be investigated in a long-term follow up study.