Designed the degenerate primers of alcohol dehydeogenase and L-lactate dehydrogenase to aug- ment Ethanoligenens harbinense B49 genomic DNA, and obtained about 780 bp and 610 bp PCR product respectively. Augmented flank sequences of the two PCR fragments with the Cassette PCR method. Similar- ity alignment showed that the products of the cloned DNA were very high similar to those of alcohol dehy- drogenase genes and L-lactate dehydrogenase genes respectively. One of the two sequences was 1902 bp long, and the ORF of adh was 1101 bp long and encoded 366 amino acids. Its putative molecular weight was about 39.71 kD, its calculational isoionic point was pH 5.93. The maximal identity and positive was 51% and 73% with Clostridium thermocellum ATCC 27405 adh. The other one was 2490 bp long, and the ORF of adh was 951 bp long and encoded 316 amino acids. Its putative molecular weight was 34.23 kD, its calcula-tional isoionic point was pH 6.09. The maximal identity and positive was 55% and 74% with Bacillus megaterium L-ldh. Successfully cloning these two genes would not only enrich the gene resources of L-lactate dehydrogenase and alcohol dehydrogenase genes, but also give the scientific warrant for the meta- bolic engineering research and the construction of the gene-engineering bacteria.

Objective Despite regular treatment,antineutrophil eytoplasmie antibodies(ANCA)asso- ciated systemic vasculitis(AASV),in which the role of Fc?Rs has been established,are still associated with significant long-term mortality and remain an important cause of end-stage renal failure.ANCA plays an im- portant role in the pathogenisis of primary systemic small vessel vasculitis(PSV)by their potential to activate neutrophils.Because the interaction between ANCA and its receptors on the Fc portion of immunoglobulins (Fc?R)on neutrophils is essential in the activation process,we investigate the inhibitory,effect of tg19320 on ANCA induced activation of neutrophils,which is a tetrameric tripeptide that interferes with IgG/Fe?Rs in- teraction.Methods We prepared tg19320 by solid-phase peptide syntbesis.The binding between tg19320 and human IgG was assessed by enzyme-linked immunosorbent assay.The biological activity of tg19320 to intefere with FcF?receptor recognition was identified by rosette formation assay.ANCA IgG was prepared from the sera of active Wegener's granulomatosis(WG)and microscopic polyangiitis(MPA)patients.Neu- trophils isolated from the blood of healthy volunteers were primed with TNF-?(2 ng/ml)and then incubated with ANCA IgG(200?g/ml),or pretreated with tg19320(2.5 mg/ml)and then added with ANCA IgG.Su- peroxide burst of neutrophils was determined by Ferri-cytochrome reduction assay.Results We found that tg19320 bound tightly to human IgG in a dose dependent manner and the inhibition of the rosette formation between SRBC-IgG and U937 cells was statistically significant(20.3% vs 53.2%,P

Objective To assess the liver histopathological characteristics of chronic hepatitis B (CHB) patients with alanine aminotransferase(ALT)lower than 2 times the upper limit of normal (ULN) through liver biopsy, and try to provide subjective evidence for clinical anti-viral treatment.Methods From October 2005 to August 2010, patients accepted liver biopsy in department of infectious disease, Sichuan provincial people's hospital were enrolled. The criteria for liver biopsy was as follow, (1) HBsAg-positive for more than 6 months, (2) HBeAg-positive patients with HBV DNA ≥103 copies/ml or HBeAg-negative patients with HBV DNA≥ 104copies/ml, (3) ALT was lower than 2 times ULN for more than 6 months,and without any hepatic protectants, (4) never accepted any antiviral treatment before, including IFN or nucleoside analogues, (5) willing to accept liver biopsy. Before liver biopsy, routine blood test, prothrombin time, liver function test, hepatitis B antigen and antibody test, HBV DNA quantification were examined. The biopsy position was located under routine ultrasound, liver biopsy were performed to assess the grading of inflammation and necrosis and the degree of fibrosis. The correlation between all the factors and liver inflammation and fibrosis were analyzed. Results Totally 383 cases (240 males and 143 females) met the diagnostic criteria, aged from 16 to 59 years old and the mean age was 28.0 years old. Cases of liver inflammation in G0, G1, G2, G3andG4 grade was 2 cases (0.5％), 165 cases (43.1％), 191 cases (49.9％), 25 cases (6.5 ％ ) and 0 cases (0 ％ ) respectively, cases≥G2 grade accounted 56.4 ％ of total. Meanwhile,stage of fibrosis in S0, S1, S2, S3 and S4 was 103 cases (26.9％), 265 cases (69. 2％), 13 cases (3.4％), 2 cases (0.5％) and 0 cases (0％) respectively, percentage of liver fibrosis in S2stage and over was only 3.9％. The occurrence of serious liver inflammation was associated with age, ALT levels, HBV DNA levels and HBeAg status (P＜0.05). There was no obvious association between HBV DNA level and liver fibrosis (P＞0.05). Conclusions There were obvious liver inflammation and different degree of liver fibrosis in CHB patients with alanine aminotransferase(ALT)lower than 2 times ULN. The degree of liver injury assessed by liver biopsys is recommended as an evaluation for the necessary of anti-viral therapy.

OBJECTIVETo investigate the value of fractional nitric oxide concentration in exhaled breath (FeNO) in assessing the level of asthma control in children.

METHODSA total of 226 asthmatic children were divided into controlled asthma (n= 86), partially controlled asthma (n=63), and uncontrolled asthma groups (n=77). Ninety healthy children were enrolled as controls. FeNO was measured for both asthmatic and healthy children using the Swedish-designed NIOX system.

RESULTSThe control group had an FeNO of 14±6 ppb, the controlled asthma group had an FeNO of 29±26 ppb, the partially controlled asthma group had an FeNO of 32±30 ppb, and the uncontrolled asthma group had an FeNO of 40±32 ppb. The three asthma groups showed significantly higher FeNO than the control group (P<0.05). The uncontrolled asthma group showed significantly higher FeNO than the controlled asthma group (P<0.05), but there were no significant differences in FeNO between the partially controlled and uncontrolled asthma groups and between the partially controlled and controlled asthma groups (P>0.05).

CONCLUSIONSAsthmatic children have significantly higher FeNO than healthy children, and FeNO is correlated with the level of asthma control.

Adolescent ; Asthma ; diagnosis ; physiopathology ; therapy ; Breath Tests ; Child ; Female ; Forced Expiratory Volume ; Humans ; Male ; Nitric Oxide ; analysis

BACKGROUNDWith economic growth and urbanization there have been significant changes in the life style and diet of urban residents in large cities of China, which is experiencing a rapid increase in the prevalence of diabetes. While high prevalence of diabetes has been reported, little is known of the long-term effects of diabetes in such a large population. The aim of this study was to estimate the morbidity rate of diabetic peripheral neuropathy (DPN) in a Chinese urban diabetic population with more than 10 years' disease duration, and evaluate the relevant risk factors. The clinical manifestation of DPN and pain status was also assessed.

METHODSFive hundred and sixty-five diabetes patients were recruited into the study. Symptoms and examination helped diagnose neuropathy. The clinical manifestation of DPN was assessed with a visual analog pain score (VAS). Diabetic complication status was determined from medical records. Serum lipids and lipoproteins, glycosylated hemoglobin (HbA1c), and the urinary albumin excretion rate were measured.

RESULTSThe morbidity rate of DPN was 46.6%. HbA1c, hyperlipidemia, and retinopathy were significantly associated with neuropathy, and these risk factors were correlated with other diabetic micro and/or macrovascular complications. The average VAS pain score of the DPN patients was 4.12 ± 2.07. Severe and moderate pain was experienced by 11.4% and 40.5% respectively of DPN patients. About 3.7% of diabetic subjects had lower limb ulcer or amputation.

CONCLUSIONSThe morbidity rate of DPN for diabetic patients with > 10 years duration is very high compared to the range reported for other populations in the world. The risk factors for DPN include HbA1c, hyperlipidemia, and retinopathy. In long-standing diabetic patients, DPN was not associated with diabetic duration, and half of the DPN patients experienced considerable daily suffering.

Aged ; China ; Diabetic Neuropathies ; epidemiology ; metabolism ; physiopathology ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Hyperlipidemias ; epidemiology ; metabolism ; physiopathology ; Male ; Middle Aged ; Pain ; etiology ; Risk Factors ; Urban Population

The safety of Chinese patent medicine has become a focus of social. It is necessary to carry out work on post-marketing clinical safety evaluation for Chinese patent medicine. However, there have no criterions to guide the related research, it is urgent to set up a model and method to guide the practice for related research. According to a series of clinical research, we put forward some views, which contained clear and definite the objective and content of clinical safety evaluation, the work flow should be determined, make a list of items for safety evaluation project, and put forward the three level classification of risk control. We set up a model of post-marketing clinical safety evaluation for Chinese patent medicine. Based this model, the list of items can be used for ranking medicine risks, and then take steps for different risks, aims to lower the app:ds:risksrisk level. At last, the medicine can be managed by five steps in sequence. The five steps are, collect risk signal, risk recognition, risk assessment, risk management, and aftereffect assessment. We hope to provide new ideas for the future research.
Clinical Trials as Topic ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; adverse effects ; chemistry ; economics ; therapeutic use ; Herbal Medicine ; economics ; legislation & jurisprudence ; Humans ; Patents as Topic ; Product Surveillance, Postmarketing ; Quality Control

OBJECTIVETo compare the liver pathohistological and clinical features between chronic HBV carriers and chronic hepatitis B patients with mild elevated in ALT.

METHODS128 patients were divided into 3 groups according to the ALT: group A: ALT is less than or equal to 0.5*ULN, group B: 0.5*ULN less than ALT is less than or equal to 1*ULN, group C: 1*ULN less than ALT less than 2*ULN. The age, sex, serum HBV DNA, HBeAg status, expression of HBcAg in liver, thickness of spleen, breadth of portal vein ,blood stream speed of protal vein, right liver obliqua diameter, grade of liver inflammation and stage of liver fibrosis were compared in the three groups.

RESULTSAmong 128 patients, 57(44.5%) patients had G1 hepatitis and 71 (55.5%) had G2 hepatitis, no G0 hepatitis was found in these patients; 72 patients (56.3%) had S1 fibrosis, 30 (23.4%) patients had S2 fibrosis, and 26 (20.3%) patients did not have liver fibrosis. The liver inflammation in group C was more aggravated than that in group A (P less than 0.05). And there were significant differences in thickness of spleen and right liver obliqua diameter between group C and group A, as well as between group C and B (P all less than 0.01). With the aggravating of liver inflammation, the serum ALT, thickness of spleen, breadth of portal vein and expression of HBcAg in liver were increased obviously (P less than 0.05). With the aggravating of liver fibrosis, the thickness of spleen, breadth of portal vein, right liver obliqua diameter and HBeAg negative patients were increased obviously, while the blood stream speed of portal vein was decreased obviously (P less than 0.01).

CONCLUSIONAmong the chronic HBV infection patients whose ALT less than 2*ULN, there were 55.5% patients had G2 of liver inflammation and 23.4% patients had S2 of liver fibrosis. The serum ALT, thickness of spleen, breadth and blood stream speed of portal vein, right liver obliqua diameter and expression of HBcAg in liver are associated with pathohistological changes in these patients.

Adult ; Alanine Transaminase ; blood ; Biopsy, Needle ; Carrier State ; blood ; pathology ; virology ; DNA, Viral ; blood ; Female ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Humans ; Liver ; metabolism ; pathology ; virology ; Liver Cirrhosis ; pathology ; Male ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Virus Replication

Armillifer agkistrodontis (Ichthyostraca: Pantastomida) is a parasitic pathogen, only reported in China, which can cause a zoonotic disease, pentastomiasis. A complete mitochondrial (mt) genome was 16,521 bp comprising 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes, and 1 non-coding region (NCR). A phylogenetic tree drawn with the concatenated amino acid sequences of the 6 conserved PCGs (atp6, cox1-3, and nad2) showed that A. agkistrodontis and Armillifer armillatus constituted a clade Pentastomida which was a sister group of the Branchiura. The complete mt genome sequence of A. agkistrodontis provides important genetic markers for both phylogenetic and epidemiological studies of pentastomids.
Amino Acid Sequence ; China ; Epidemiologic Studies ; Genes, rRNA ; Genetic Markers ; Genome ; Genome, Mitochondrial* ; Humans ; Pentastomida* ; RNA, Transfer ; Siblings ; Tongue* ; Trees ; Zoonoses

OBJECTIVETo explore the therapeutic feasibility of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from partially HLA-mismatched related donors for hematologic diseases.

METHODSThirty patients with hematologic diseases received allo-HSCT from 1 - 3 loci mismatched related donors conditioning regimen consisting of ATG (thymoglobulin, total dose of 10 mg/kg, intravenously on - 4 d to - 1 d), and only 5 (18%) of 28 recipients from HLA-identical sibling donors were treated with regimen containing ATG. Donors were given G-CSF prior to hematopoietic stem cell harvest and CsA, short-term MTX and mycophenolate mofetil (MMF) were used for GVHD prophylaxis in both group.

RESULTSAll patients were successfully engrafted. There was no significant difference in the incidence of grade II to IV acute graft-versus-host disease (aGVHD) and grade III to IV aGVHD between the mismatched and matched groups (34% vs 32%, and 13% vs 11%, respectively). 3-year overall survival (OS) and disease-free survival (DFS) in mismatched and matched groups were 57% vs 77% (P = 0.14) and 57% vs 69% (P = 0.28), respectively. Multivariate analysis showed that advanced disease pre-transplant (P = 0.006) and CMV infection (P = 0.04) were risk factors for OS. OS for patients with stable disease in mismatched and matched groups were 87% vs 81% (P = 0.65) respectively, and for those with advanced disease were 21% vs 71% (P = 0.02).

CONCLUSIONSIt is feasible to perform allo-HSCT from 1 -3 loci HLA-mismatched related donors for patients with stable disease who lack HLA-identical sibling donors. Nevertheless, for patients with advanced disease optimized conditioning regimen and intensive supporting therapy should be administered to obtain better clinical outcomes.

Graft vs Host Disease ; prevention & control ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Siblings ; Tissue Donors ; Transplantation Conditioning

To investigate the relationship between the plasma levels of chemokine CXCL9/Mig and acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The plasma levels of CXCL9/Mig of 35 patients who received all-HSCT were detected by using ELISA assay, these patients included 13 patients with grade 0-I, 12 patients with grade II and 10 patients with grade III - IV aGVHD, respectively. The four different time points including prior to allo-HSCT, one week before aGVHD onset, the plateau of aGVHD and time after completely controlled, were studied. The results showed that the plasma levels of CXCL9/Mig in the patients with serious aGVHD (grade II - IV) were significantly increased during aGVHD than those in the patients without aGVHD or with slight aGVHD (P < 0.001). It was found that CXCL9/Mig levels were significantly correlated with the severity of grade aGVHD (P < 0.001). Another important finding was that CXCL9/Mig levels obviously increased at one week before aGVHD was diagnosed. CXCL9/Mig level was not obviously correlated with CMV infection or other infectious complication (P > 0.05). It is concluded that the plasma level of CXC19/Mig significantly correlated with the severity of aGVHD and plays a critical role in pathogenesis of aGVHD, the changes in plasma level of CXCL9/Mig after allo-HSCT may be used as a valuable indicator for early diagnosis of aGVHD, finally, provide a early therapeutic approach to reduce aGVHD severity and improve the outcome for patients after allo-HSCT.
Chemokine CXCL9 ; blood ; Graft vs Host Disease ; blood ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans