Objective To study the possibility of using heterogeneous antiserum in single radial immunodiffusion (SRID) for quantitative detection of HA in H7N9 influenza vaccine product when H7N9-specific antiserum is not available in order to establish a testing method for the detection of H 7N9 antigen in any urgent situation.Methods Antisera specific for H7N1, H7N2, H7N3 and H7N7 were obtained from NIBSC and used for SRID assay .Amino acid sequences of hemagglutinins were comparatively analyzed be-tween H7N9 virus and other viruses used to prepare heterogeneous antiserum .The titers of antisera against H7N9 and their homogenous antigens were detected by double immunodiffusion method .Based on the results of homology analysis and cross-reaction, a suitable antiserum was selected and its applicability was further validated by the SRID assay using H7N9 antigen.Results Influenza A virus subtype H7N3 that used for preparation of 07/278 antiserum showed the highest HA homology with H7N9 (97.14%).The titer of 07/278 antiserum against H7N9 antigen was 1 ∶8 as detected by double immunodiffusion assay .The H7N9 anti-gen and the 07/278 antiserum could form a clear precipitation line in SRID assay .The detection of H7N9 antigen in the range of 10 to 40μg/ml showed a good linearity in the standard curve .Conclusion The 07/278 antiserum from NIBSC can be used as an alternative reagent for the quantitative detection of hemaggluti -nin in H7N9 influenza virus vaccine .

A total of 9 806 type 2 diabetics managed by the communities were selected by the stratified cluster random sampling.The characteristics, behavior and life style, history of diseases and treatments, and familial history were collected by a standard questionnaire.Their heights and weights were measured.Furthermore, their HbA1C was tested.Logistic regression was used to analyze the association between familial history of diabetes and glycemic control.The results showed that among the diabetics, patients with familial history accounted for 18.99%, and glycemic control rate was 42.72%.Compared with the diabetics without familial history, glycemic control rate in patients with parental history of diabetes and with many relatives decreased by 0.27 fold (OR=1.27, 95%CI 1.01-1.59) and 1.01 fold (OR=2.01, 95%CI 1.25-3.23), suggesting that family history of diabetes could reduce the glycemic control rate.

Objective To explore the protective effects and its mecha-nism of stachydrine on focal cerebral ischemia reperfusion injury in mice . Methods The focal cerebral ischemia reperfusion injury model in mice was established by thread method.KM mice were randomly divided into 6 groups(n＝16):sham operation group , model group, control group, ex-perimental-L, experimental -M, experimental -H groups, mice in each group were given related drugs via intragastric administration before the operation, once a day, lasting for 7 d.Reperime 22 h, the levels of Caspase-3, B-cell lymphoma-2 (Bcl-2), B-lymphoma-2 gene-related X protein (Bax) and adenosine triphosphate (ATP) enzyme activity in each group was detected by enzyme linked immunosorbent assay .Results The relative expression of Bax in model group,control group, experimental -L, experimental -M, experimental -H groups were (4.00 ±0.51 ), (2.84 ±0.48),(2.87 ±0.41),(2.92 ±0.46),(3.12 ±0.59)ng· mL-1,respectively; the relative expression of Caspase-3 in the 5 groups were(13.43 ±0.87),(9.95 ±1.57),(10.02 ±1.55),(10.58 ±1.48),(10.79 ±1.69) pmol· L-1,respectively; the relative expression of Bcl -2 in the 5 groups were (11.94 ±1.40 ),(16.14 ±1.33 ), (15.89 ±1.45),(15.31 ±1.20),(14.78 ±1.17)ng· mL-1,respectively;compared with the model group, the rela-tive expressions of Bax and Caspase -3 in control group, three doses experimental groups markedly reduced while the relative expression of Bcl-2 in the 4 groups markedly increased significantly (all P<0.01).The Na +-K+-ATP en-zyme activity in the model group ,control group, experimental -L, experimental -M, experimental -H groups were (2.82 ±0.56),(4.84 ±1.61),(4.81 ±1.48),(4.50 ±1.22),(4.13 ±1.17)μmolPi· mg -1· h -1,respectively;and Mg2+-ATP enzyme activity in the 5 groups were (2.55 ±0.56), ( 5.25 ±1.86 ), ( 5.15 ±2.37 ), (4.62 ±1.89 ) ,(4.13 ±1.54 ) μmolPi· mg-1· h-1, respectively; comparison between 4 drugs groups and model group,the differences of the factors were significant ( P<0.05 or P<0.01 ) .Conclusion Stachydrine can markedly reduce the lesion of focal cerebral ischemia reperfusion model in mice , and its mechanism may be related to improve energy metabolism disorder , inhibit neuronal apoptosis and improve the microcirculation of the brain .

OBJECTIVETo investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.

METHODSHistological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.

RESULTSCompared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].

CONCLUSIONSAbnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.

Aged ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Female ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Skin ; metabolism ; pathology ; Wound Healing

OBJECTIVETo evaluate the usefulness of new microspincolumn method for the measurement of a1pha-fetoprotein variant AFP-L3 in differentiation of benign and malignant liver disease and the warming for liver cancer.

METHODSAFP-L3 was isolated by using microspincolumn coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were determined with chemiluminescent immunoassay, the proportion of AFP-L3 levels AFP-L3(%) were calculated, and the relationship between the elevated AFP-L3(%) levels and benign and malignant liver disease was analyzed.

RESULTSThe levels of AFP-L3(%) in serum of patients with hepatocellular carcinoma was significantly higher than those in the patients with other liver diseases (P < 0.001). Taking AFP-L3(%) >or= 10% as the diagnostic criteria, the sensitivity for diagnosis of liver cancer was 90.9%.

CONCLUSIONDetection of AFP-L3 seemed to be of clinical value in diagnosis and differential diagnosis of hepatocellular carcinoma; it may be especially important for identifying patients with hepatocellular carcinoma whose a1pha-fetoprotein level is low.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Diagnosis, Differential ; Female ; Hepatitis, Chronic ; blood ; diagnosis ; Humans ; Immunoassay ; methods ; Liver Cirrhosis ; blood ; diagnosis ; Liver Neoplasms ; blood ; diagnosis ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Administration, Oral ; Animals ; Arylamine N-Acetyltransferase ; genetics ; metabolism ; Caffeine ; metabolism ; urine ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Medicine, Tibetan Traditional ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Theophylline ; urine ; Uracil ; analogs & derivatives ; urine ; Uric Acid ; analogs & derivatives ; urine ; Xanthines ; urine

Background Since the measurement method establishment of serum ferritin abroad in early period of theseventies, the iron deficiency had been divided into two types: the non-anemia and anemia types. In orderto go step further studies, we must ertablish the bemoglobin targets of the two types. Methods One hurdred and fifty-two children in experimontal group, from 6 to 7 years old, and allcome from Qinghai province. There are 29 children in Xining city, 24 in Guide, 26 in Gongbe, 40 in Gui-nan and 33 in Maduo countics. There are 36 health children aged from 6 to 7 years old in the controlgroup, and all comes from Beijing. The Hb, RBC, HCT, HCTW and FEP wcre determined. Results The three targets correlating with Hb (Hb, MCH and MCHC); correlating with RBC (RBC,HCT and MCV); the two targets correlating with RBC_weight (HCTW and CMCW) and correlating withFEP of RBC(FEP and MCEP) have very significant difference between experimental group and control group. Conclusion The determination values of the 10 targets are not same in children in different districts,and the values of all the target: are increased on different degree along with the increase in altitude of ele-vation. There is very important significance on the studies of iron deficiency and altitude hypoxia to establish the normal values of the 10 targets.

AIM:To explore the role of aloperine in ischemia-reperfusion(I/R)-induced H9c2 cardiomyocyte injury and inflammation.METHODS: The H9c2 cardiomyocytes were cultured under hypoxia and re-oxygenation condi-tions to simulate ischemia-reperfusion(SI/R)injury.After treatment with aloperine at various doses,the cell viability was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.Simultaneously,the levels of lactate dehydro-genase(LDH),malonaldehyde(MDA)and caspase-3 activity were detected by the commercial kits.The levels of inflam-matory cytokines were also detected by ELISA.Moreover,the effects of aloperine on the activation of PI 3K/AKT signaling pathway were determined by Western blot.RESULTS:Pre-treatment with aloperine remarkably abated the inhibitory effect of SI/R on H9c2 cell viability,and decreased the elevations of LDH and MDA triggered by SI /R(P<0.05).Pre-treat-ment with aloperine dramatically suppressed the cell apoptosis induced by SI /R treatment(P<0.05), concomitant with the decrease in caspase-3 activity and increase in Bcl-2/Bax ratio(P<0.05).In contrast to SI/R group,aloperine treat-ment notably restrained the concentrations of pro-inflammatory cytokines, including interleukin-6, tumor necrosis factor-α and interleukin-1β(P<0.05).Furthermore, aloperine remarkably increased the protein levels of p-PI3K and p-AKT. While blocking the PI3K/AKT pathway with its specific inhibitor LY294002, the viability-promoting, anti-apoptotic and anti-inflammatory effects of aloperine on the H 9c2 cells were obviously attenuated(P<0.05).CONCLUSION: Alope-rine protects against cardiomyocytes from I/R injury and inhibits inflammatory responses by activating the PI 3K/AKT signa-ling pathway,implying a potential benefic role of aloperine against myocardial I /R injury.

In recent years,the prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKP)infection has become a global public health issue.Ceftazidime/avibactam(CAZ/AVI)has been approved as a novel antimicrobial agent for the treatment of healthcare-associated pneumonia/ventilator-associated pneumonia,bloodstream infection,infection after kidney transplantation,and severe infection combined with liver cirrhosis.However,the use of CAZ/AVI has also led to the emergence of drug-resistant strains.The major mechanisms of drug-resistance include over-expression of blaKPC gene,mutation of β-lactamase and amino acids at key sites,changes in cell permeability caused by loss of membrane porin,and over-expression of efflux pump.This article reviews the research progress of CAZ/AVI in the treatment of CRKP infection,providing reference for clinical diagnosis and treatment.

OBJECTIVETo investigate the effects of excessive intake of fluoride on the expression of type II collagen gene and types and morphological change of collagen fiber in the bone tissues of rats.

METHODSA rat model with fluorosis was established by adding 221 mg/L of sodium fluoride (NaF) to drinking water for the rats for 15 days, 30 days and two months, respectively. Type II collagen alpha1 (II) cDNA probe was prepared, and cDNA-mRNA in-situ hybridization was employed to detect change in expression of type II collagen mRNA in the bone tissues of rats with excessive intake of fluoride (221 mg/L NaF). Picrosirius-polarization method was used to observe types of collagen and morphology of collagen fiber in the bone tissues.

RESULTSChondroblasts were found in the femur and other bone tissues of the rats after exposure to fluoride. cDNA-mRNA in-situ hybridization showed that expression of type II collagen gene could be observed in the cytoplasm of chondrocytic lacuna and chondrified bone tissues. mRNA in collagen of chondrocytes of the rib cartilage reached the peak level 15 days after exposure to fluoride, and decreased gradually one month and two months after exposure. Polychromatic type II collagen, breakage of collagen fiber, disorder array and reduced content of type II collagen could be found in the bone tissues with picrosirius-polarization method.

CONCLUSIONSExcessive intake of fluoride could lead to changes in types and structure of collagen (cross-linkage) of bone tissues, which caused expression of type II collagen gene in the chondrified bone tissues and enhanced its expression in the rib cartilage tissues.

Animals ; Bone Diseases ; metabolism ; pathology ; Chondrocytes ; metabolism ; Collagen Type II ; biosynthesis ; genetics ; Fluoride Poisoning ; genetics ; metabolism ; pathology ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar