Background Research demonstrated that mitochondrial pathway plays a key role in cell apoptosis.Purendan supermicropowder(PRD),a traditional Chinese medicine,may be a potentially effective therapy for neuron apoptosis in diabetic retina.Objective This study was carried out to investigate the effects of PRD on aldose reductase(AR)activity,neuron apoptosis and mitochondrial pathway in retina of diabetic rat.Methods Thirty-six clean male Wistar rats were randomly divided into normal control group,diabetes model group,PRD treatment group randomly and 12 rats for each group.The diabetes models were established by intraperitoneal injection of 25 mg/(kg · d)streptozotocin(STZ)for 3 consecutive days,and blood glucose ≥ 16.7 mmol/L was taken as the standard.PRD solution of 1.8 g/(kg · d)was lavaged in 12 models for 3 months.The eyeballs were enucleated for the preparation of retinal tissue homogenate and slice.AR activity in the retina was detected by ultraviolet spectrophotometry,and neuron apoptosis in retina was assayed by TUNEL staining.Western blot was used to assess the expressions of bcl-2,bax,cyt-c and caspase-3 protein in the retina.The use of animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Committee(Version 1988).Results Statistically significant differences were found in AR activity and AI among the normal control group,diabetic group and PRD groups(F=90.115,165.540,P＜0.01),and those of diabetic group were evidently higher than the normal control group and PRD group(P＜0.01,P＜0.01).The positive TUNEL cells mainly located in inner nuclear layer and retinal ganglion cell layer.The expressions of bax,cyt-c,caspase-3,bcl-2 and bcl-2/bax in retina were obviously different among these three groups(F =51.332,41.262,25.888,38.564,47.870,P＜0.01),and the expression of bax,cyt-c and caspase-3 protein in diabetic group evidently elevated in comparison with the normal control group and PRD group(t ＝ 10.32,11.04,6.91,P ＜ 0.01)and the expressions of bcl-2 protein and bcl-2/bax value were significantly lower in diabetic rats than in the normal control rats(t =18.05,12.23,P＜0.01).AR activity by AI of retina,the expressions of bax,cyt-c and caspase-3 proteins in retina were obviously lower in PRD group than in diabetes model rats(P ＜ 0.01),and the expression of bcl-2 protein and bcl-2/bax value were significantly higher in PRD group than in diabetes group(P＜0.01).Conclusions PRD can protect retina against the damage caused by high glucose by suppressing AR activity by downregulating the expressions of bax,cyt-c,caspase-3 proteins,increasing the expressions of bcl-2 protein in retina of diabetic rats and further inhibiting the mitochondrial pathway and reducing cell apoptosis in retina of diabetic rats.

OBJECTIVETo investigate the relationship between penA/ponA and penicillin resistance of Neisseria gonorrhoeae.

METHODSAgar dilution method was used to determine the minimum inhibitory concentrations (MICs) of the strains. Polymerase chain reaction-single stand conformation polymerphism (PCR-SSCP) and PCR- restriction fragment length polymorphism (RFLP) were used to detect the mutations in ponA and penA genes, which encoding the penicillin binding protein-1 and -2 (PBP1 and PBP2), respectively.

RESULTSAll the 80 N. gonorrhoeae isolates had a D345 insertion detected in penA while 93.7% of N. gonorrhoeae isolates having a point mutation Leu421 --> Pro in ponA. Most of the penicillinase producing N. gonorrhoeae (PPNG) strains possessed the mutations in ponA and penA.

CONCLUSIONOur data suggested that the plasmid and chromosome mediated penicillin-resistance conjugately increased the level of resistance.

Amino Acid Sequence ; Bacterial Proteins ; genetics ; Base Sequence ; DNA Mutational Analysis ; DNA, Bacterial ; Genes, Bacterial ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Mutagenesis, Insertional ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; Penicillin Resistance ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational

In the late part of the 20th century, due to the change of disease spectra, rapid increase of health expenditure and imbalanced distribution of health resources, the health management sectors and doctors increasingly demand the scientific decision-making. With the development of clinical evidences and research, evidence-based medicine (EBM) became emerged in the early 1990's. EBM, the medical science in compliance with the best available clinical evidence, integrates clinical epidemiology (CE), systematic reviews (SRs) and health technology assessment (HTA) as the major technology, to emphasize that the clinical practice should be based on the combination of the best available clinical evidence, medical experiences and patient desire to cope with the demand of medical decision-making. Some governments and international organizations like WHO, and Australia have now adopted the high-quality evidence of EBM for decision-making. As the largest developing country, China is challenged by variety of health problems. The introduction, adaptation and implementation of EBM will improve the decision-making and health care services, which should enhance the competitive capacity of medical and pharmaceutical manufactures, as well as promote the popularization of EBM, and protect the doctors and patients rights.
Epidemiologic Methods ; Evidence-Based Medicine ; Forecasting ; Health Policy ; trends ; Humans ; Meta-Analysis as Topic ; Policy Making

Objective From May 2009-January 2010,a total of 3768 biosamples were tested for influenza A ( H1N1 ) infection at Zhengzhou center for disease control and prevention,China.1452 cases were laboratory confirmed H1N1 infection and 2316 were considered suspected victims.To evaluate the current protocol of influenza A (H1N1) based on the epidemic situations of Zhengzhou,relationships among features were explored and whether additional clinical characteristics should be part of H1N1 diagnosis protocols were determined.Methods Both clinical and epidemiologic findings as well as statistical analyses were described in this article.Test for independence between features related to the disease diagnosis has been proposed.Furthermore,logistic regression was carried out to measure the association among features and latent class analysis was performed to identify additional crucial features in laboratory confirmed H1N1 by building various latent models with different combinatorial features.Results The mean generation time for H1N1 was estimated as 3.59 ± 1.41 days (range =2.01 -7.26).The estimated infection rate was 0.258 ±0.088 3,and reproduction number was 1.94 (95％ CI =1.12 - 3.18 ).Our results revealed that the six features,including molecular detections using three separate primer/probe sets,gender,age and temperature,are all associated with clinical diagnosis of H1N1,and that three separate primer/probe sets for laboratory confirmed H1N1,age and temperature are associated with each other.Conclusion Additional clinical features applied into the H1N1 diagnosis with current three primers/probe sets can increase the diagnostic efficiency.

OBJECTIVEUsing molecular epidemiology methods to investigate relationship between genotypes and drug-resistance of neisseria (N.) gonorrhoeae in Shanghai area.

METHODSA random amplified polymorphic DNA (RAPD) fingerprint method at the molecular level was used to differentiate the strains which were isolated from the outpatients of sexually transmitted disease clinics. The sensitivity to antibiotic of the 78 N. gonorrhoeae strains on 9 different antibiotics was tested and the relationship between different genotypes and phenotypes was studied.

RESULTSSelected RAPD primer could give out a group of amplification polymerase chain reaction bands with some main segments common to all the N. gonorrhoeae strains tested and some segments were different among the N. gonorrhoeae strains. All the 78 N. gonorrhoeae strains could be classified as three different groups (I, II and III). The strains could also be distinguished as four types (A, B, C and D) according to drug-resistance status. Using correspondence analysis method, the relationship between the three genotypes and four resistance types could be identified.

CONCLUSIONRAPD fingerprint seemed a useful genotyping method and could be used for molecular epidemiological studies.

Adolescent ; Adult ; Anti-Bacterial Agents ; pharmacology ; China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Female ; Genotype ; Gonorrhea ; epidemiology ; microbiology ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Molecular Epidemiology ; Neisseria gonorrhoeae ; classification ; drug effects ; genetics ; Random Amplified Polymorphic DNA Technique

BACKGROUNDThe pathogenesis of acute pancreatitis is complex and largely unclear. The aim of this study was to explore the relationship between modes of cell death in pancreatic acinar cells, the release of cell contents and the inflammatory response of macrophages.

METHODSOur experiment included four groups: group A (the control group), group B (AR42J cells overstimulated by caerulein), group C (AR42J cells treated with lipopolysaccharide and caerulein), and group D (AR42J cells treated with octreotide and caerulein). Apoptosis and oncosis, and the release of amylase and lactate dehydrogenase (LDH) from AR42J cells were detected. Rat macrophages were stimulated by 1 ml supernatant of culture medium of AR42J cells. Finally, NF-kappaB activation and TNF-alpha and IL-1beta secretion by macrophages were detected.

RESULTSOncotic cells in group C increased while apoptotic cells decreased (P < 0.05); cells in group D had the inverse reaction. The release of amylase and LDH changed directly with the occurrence of oncosis. The transcription factor NF-kappaB was activated and secretion of TNF-alpha and IL-1beta were significantly higher in group C than in group B (P < 0.05); in group D, these actions were significantly lower than in group B (P < 0.05). This trend was in line with changes in amylase and LDH production.

CONCLUSIONThere is a close relationship between modes of pancreatic acinar cell death, the release of cell contents and the inflammatory reaction of macrophages.

Amylases ; secretion ; Animals ; Apoptosis ; Interleukin-1beta ; secretion ; L-Lactate Dehydrogenase ; secretion ; Macrophage Activation ; Male ; NF-kappa B ; metabolism ; Pancreas ; pathology ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; secretion

Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blue populations and the correlation with gastromedicine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.
Base Sequence ; Benzyl Alcohols ; analysis ; Cloning, Molecular ; Computational Biology ; DNA, Plant ; chemistry ; Gastrodia ; genetics ; Glucosides ; analysis ; Plant Tubers ; genetics

OBJECTIVETo evaluate the safety and efficacy of needlescopic thoracic sympathectomy for palmar hyperhidrosis.

METHODSFrom March 2004 to April 2005, 62 patients, including 23 men and 39 women with a mean age of 23 years (ranged from 12 to 53 years), underwent bilateral needlescopic thoracic sympathectomy. Among all the patients 8 cases had accompanied axillary hyperhidrosis. The degree of palmar sweating was moderate in 16 cases and severe in 46 cases. The sympathetic chain on the body of the second and third ribs in all patients was cauterized and divided; the chain on the fourth rib in those with axillary hyperhidrosis was also severed. Intraoperative changes in palmar temperature and blood flow were recorded.

RESULTSSympathectomies were successful, and dry limbs were immediately achieved in all patients after surgery. There were no mortality or life-threatening complication, however 1 patient developed moderate pneumothorax which resolved soon after chest drainage. After all procedures, palmar blood perfusion increased significantly and mean palmar temperature elevated by 2.4 degrees C. The mean operative duration was 65 min, and the mean postoperative hospital stay was 1.2 days. No recurrence of palmar hyperhidrosis occurred after a mean follow-up of 6.3 months (ranged from 1 to 13 months). Compensatory sweating was found in 26 patients, but the symptoms were mostly tolerable and required no further treatment.

CONCLUSIONNeedlescopic thoracic sympathectomy is a safe and effective technique for palmar hyperhidrosis, which is less invasive than conventional video-assisted thoracic surgery.

Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Hand ; Humans ; Hyperhidrosis ; surgery ; Male ; Middle Aged ; Sympathectomy ; methods ; Thoracic Surgery, Video-Assisted ; Treatment Outcome

OBJECTIVETo analyze the clinical features and prognosis of the primary myelodysplastic syndrome with myelofibrosis (MDS-MF) patients and to improve the cognition of MDS-MF.

METHODSFour hundred and sixty-six primary MDS patients with bone marrow (BM) biopsy were divided into two groups according to whether BM associated with fibrosis, the clinical features and prognosis of the two groups were analyzed retrospectively.

RESULTS167 (35.8%) MDS cases revealed myelofibrosis, of which MF-1 123 cases (26.4%), MF-2 40 cases (8.6%), MF-3 4 cases (0.9%). The proportion of hepatosplenomegaly in MDS-MF group was significantly higher than in MDS without MF group, the difference had statistical significance (P = 0.031). The proliferation of BM biopsy in MDS-MF group was significantly more active than in MDS without MF group. The number of blasts, megakaryocytes and abnormal megakaryocytes in MDS-MF group were significantly higher than in MDS without MF group, the differences had statistical significance (P < 0.05). Among the 345 patients who had available results of cytogenetic analysis, 121 cases were MDS-MF patients, the proportion of middle and high-risk prognostic group according to IPSS karyotype prognosis groups in MDS-MF group were significantly higher than in MDS without MF group, the differences had statistical significance (P = 0.047). The median survival was 17 (1 - 60) months in MDS-MF group, and was 32 (1 - 62) months in MDS without MF group. The difference had statistical significance (P = 0.001). Myelofibrosis had independent prognostic significance by multi-variable analysis (P = 0.019).

CONCLUSIONThe myelofibrosis in MDS is main the proliferation of reticular fiber. The proliferation of reticular fiber is closely related with the number of blast cells, the proliferation and developmental abnormalities of megakaryocytes and the karyotype. The prognosis of MDS-MF patients is poor.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; diagnosis ; pathology ; Primary Myelofibrosis ; complications ; diagnosis ; pathology ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo explore the frequencies of heterozygosity in X-linked G6PD, P55, BTK, and FHL-1 gene exonic polymorphic loci among Chinese females and the value of determination of hematopoietic clonality by detection of these X-chromosome exonic polymorphisms based on X-chromosome inactivation patterns (XCIP)-transcription-based clonality assays (TCA).

METHODSGenomic DNA was extracted from peripheral blood of 446 Chinese healthy females. Allele-specific PCR (ASPCR) or PCR-restriction enzyme digestion method was applied for detecting G6PD, P55, BTK and FHL-1 polymorphisms. Those heterozygotic loci were used as markers to examine the hematopoietic clonality of bone marrow mononuclear cells by TCA from essential thrombocythemia (ET) patients with JAK2V617F mutation and myelodysplastic syndrome (MDS) patients with abnormal karyotype.

RESULTSAmong the total 446 genomic DNA samples, the frequencies of heterozygosity in G6PD, P55, BTK and FHL-1 loci were 12.8%, 29.4%, 52.0% and 46.4%, respectively. About 81.4% of females were heterozygous at one or more loci. All 10 ET patients with JAK2V617F mutation and 2 MDS patients with abnormal karyotype, which were heterozygotic in either locus, had monoclonal/oligoclonal hematopoiesis.

CONCLUSIONClonality detection based on X chromosome inactivation patterns-transcription based clonality assays is applicable to about 80% of Chinese females.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Chromosomes, Human, X ; Exons ; Female ; Genes, X-Linked ; Genetic Carrier Screening ; Genetic Linkage ; Glucosephosphate Dehydrogenase ; genetics ; Hematopoiesis ; genetics ; Humans ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; X Chromosome Inactivation ; Young Adult