Research on novel pullulanase has major significance on the domestic industrialization of pullulanase and the breakdown of foreign monopoly. A thermophilic bacteria LM 18-11 producing thermostable pullulanase was isolated from Lunma hot springs of Yunnan province. It was identified as Anoxybacillus sp. by 16S rDNA phylogenetic analysis. Full-length pullulanase gene was cloned from Anoxybacillus sp. LM18-11. The optimum temperature of the pullulanase was between 55 and 60 degrees C with a half-life as long as 48 h at 60 degrees C; and its optimum pH was between 5.6 and 6.4. V(max) and K(m) of the pullulanase was measured as 750 U/mg and 1.47 mg/mL, which is the highest specific activity reported so far. The pullulanase crystals structure showed a typical alpha-amylase family structure. The N-terminal has a special substrate binding domain. Activity and substrate binding were decreased when the domain was deleted, the V(max) and K(m) were 324 U/mg and 1.95 mg/mL, respectively. The pullulanase was highly heterologous expressed in Bacillus subtilis by P43 promoter. The extracellular enzyme activity was 42 U/mL, which increased more than 40 times compared to the initial strain. This pullulanase has good application prospects.
Anoxybacillus ; classification ; enzymology ; China ; Glycoside Hydrolases ; metabolism ; Hydrogen-Ion Concentration ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Temperature

Bronchi ; abnormalities ; pathology ; Bronchial Diseases ; complications ; congenital ; diagnosis ; pathology ; Bronchoscopy ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Infant ; Male ; Pneumonia ; etiology ; physiopathology ; Respiratory System Abnormalities ; complications ; diagnosis ; pathology ; Trachea ; abnormalities ; pathology ; Tracheal Stenosis ; etiology ; pathology

OBJECTIVE: To establish the linear regression equation between body height and combined length of manubrium and mesostenum of sternum measured by CT volume rendering technique (CT-VRT) in southwest Han population. METHODS: One hundred and sixty subjects, including 80 males and 80 females were selected from southwest Han population for routine CT-VRT (reconstruction thickness 1 mm) examination. The lengths of both manubrium and mesosternum were recorded, and the combined length of manubrium and mesosternum was equal to the algebraic sum of them. The sex-specific linear regression equations between the combined length of manubrium and mesosternum and the real body height of each subject were deduced. RESULTS: The sex-specific simple linear regression equations between the combined length of manubrium and mesostenum (x3) and body height (y) were established (male: y = 135.000+2.118 x3 and female: y = 120.790+2.808 x3). Both equations showed statistical significance (P < 0.05) with a 100% predictive accuracy. CONCLUSION CT-VRT is an effective method for measurement of the index of sternum. The combined length of manubrium and mesosternum from CT-VRT can be used for body height estimation in southwest Han population.
Asian People ; Body Height ; Female ; Forensic Anthropology ; Humans ; Linear Models ; Male ; Manubrium/anatomy & histology* ; Regression Analysis ; Sternum/anatomy & histology* ; Tomography, X-Ray Computed

Child ; China ; Community-Acquired Infections ; microbiology ; Drug Resistance, Bacterial ; genetics ; Haemophilus influenzae type b ; drug effects ; genetics ; isolation & purification ; Humans ; Microbial Sensitivity Tests ; Pneumonia ; microbiology

Objective To assess the feasibility of three-dimensional contrast-enhanced ultrasonic cholangiography(3D-CEUSC) in cadaver liver.Methods The 3D-CEUSC was performed in 6 cases of cadaver liver.Image quality of 3D-CEUSC was evaluated.The visualization of branching orders,the degree of visibility and coincidence of morphous were compared with those of cholangiagraphy using fluoroscopy.Results The imaging quality of 3D-CEUSC was inferior to that of cholangiography with significant difference.The three-dimensional biliary tree structures were visualized in all 6 3D-CEUSC.The maximum visualization of branching orders in 3D-CEUSC was (3.67±0.52),which was equal to the results(4.00±0.63)by cholangiography (P=0.465).The degrees of visibility of biliary tree were equivalent with those by cholangiography in the first and second order with significant difference.The coincidence of morphous was excellent compared the images of 3D-CEUSC with direct X-ray cholangiography.Conclusion 3D-CEUSC is a new technique as a useful supplement to cholangiography in evaluation of biliary anatomy and variation before graft harvesting in LDLT.

Hypertension is one of main risk factors for the occurrence and death of stroke and coronary heart disease. Its prevalence rate is rising year by year. It severely threatens the health of the human beings. The acupuncture method of "activating blood and dispersing wind, harmonizing Gan-Pi" for treating hypertension launched by Academician SHI Xue-min has aroused great attention due to good cur- ative effect and less adverse reactions. In this paper principles of the circular motion covered by the acupuncture method of "activating blood and dispersing wind, harmonizing Gan-Pi" were clarified.
Acupuncture ; Acupuncture Therapy ; methods ; Coronary Artery Disease ; Humans ; Hypertension ; therapy ; Medicine, Chinese Traditional ; Risk Factors ; Stroke ; Wind

OBJECTIVETo investigate the inhibitory effects of humanized monoclonal antibody-3 (huTNT-3) mediated truncated tissue factor (tTF) on the H(22) hepatoma-bearing mice, and to explore its mechanisms.

METHODSThe coagulation activity of the huTNT-3/tTF fusion protein was detected by clotting assay and clotting factor X (FX) activation test in vitro. Mouse hepatoma cell line H(22) cells were inoculated subcutaneously into mice to establish the mouse models of hepatoma. The mice were randomly divided into two groups to be injected once with huTNT-3/tTF fusion protein or tTF protein labeled with rhodamine B isothiocyanate (RBITC), respectively. The localization of huTNT-3/tTF fusion protein in the mouse hepatoma tissue was analyzed by confocal laser scanning microscopy 24 hour after the injection. Fifteen mice were randomly divided into three groups to be injected with the huTNT-3/tTF fusion protein, tTF protein or phosphate buffered saline (PBS) once, respectively. The tumor size was measured every two days to calculate the tumor volume. Ten days after the injection the mice were sacrificed. Samples of the tumor, heart, livers, spleen, lung, kidney and brains of the mice were taken for histopathological examination.

RESULTSBoth the huTNT-3/tTF fusion protein and tTF protein effectively promoted blood coagulation. Under the conditions of Ca(2+), the coagulation time in the 1.5, 3, 6 µmol/L huTNT-3/tTF groups was (12.90 ± 0.60) min, (10.39 ± 0.40) min and(8.15 ± 0.24) min, respectively, and the coagulation time of the 1.5, 3, 6 µmol/L tTF groups was (14.23 ± 0.46) min, (12.10 ± 0.49) min and (9.83 ± 0.52) min, respectively, the difference between the two groups was not significant (F = 0.145, P = 0.705). The huTNT-3/tTF fusion protein was similar to the tTF protein in the ability of activating FX (t = 0.101, P > 0.05). The confocal laser scanning microscopic analysis showed that RBITC-fluorescence labeled huTNT-3/tTF fusion protein was enriched in the hepatoma tissue. The tumor volume of the huTNT-3/tTF fusion protein group was significantly lower than that of the tTF and PBS groups (both P < 0.001), however, there was not significant difference between the tTF and PBS groups (t = -0.616, P > 0.05). The survival time of the huTNT-3/tTF group was (25.5 ± 2.5) d, significantly longer than that of the PBS group (17.3 ± 1.9) d and the tTF group (18.6 ± 1.9) d, (both P < 0.05).

CONCLUSIONThe huTNT-3/tTF fusion protein retains the coagulation ability and has the capability of targeting to tumor vasculature, and induces thrombosis in the tumor vessels, thus to suppress the growth of hepatoma in the mice.

Animals ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Blood Coagulation ; Carcinoma, Hepatocellular ; blood ; pathology ; therapy ; Cell Line, Tumor ; Factor X ; metabolism ; Liver Neoplasms ; blood ; pathology ; therapy ; Male ; Mice ; Neoplasm Transplantation ; Random Allocation ; Recombinant Fusion Proteins ; therapeutic use ; Thromboplastin ; therapeutic use ; Tumor Burden

Objective To explore the possibility of basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)combined with striatal conditioned medium promoting the directional differentiation of rat bone marrow mesenchymal stem cells(BMMSCs)into dopaminergie neurons.Methods 1.Separation and culture of BMMSCs:BMMSCs were harvested from healthy adult Wistar rats for serial subcultivation.2.Preparation of Striatal conditioned medium:newborn Wistar rats within 24 hours were selected,and their brain tissues were removed to prepare striatal conditioned medium.3.Induced differentiation of BMMSCs:the 5th passage BMMSCs were collected and pre-induced in low glucose-Dulbecco's modified eagle medium(L-DMEM)containing bFGF and EGF.Twenty-four hours later,pre-induction liquor was replaced with striatal conditioned medium for further induced differentiation.4.Result assessment:the morphological changes of stem cells were observed under inverted phase microscope.The expression of neuron specific enolage(NSE)and tyrosine hydmxylage(TH)were identified by immunocytochemical technique.Results The cell body of rat BMMSCs contracted into round and spindle shape after induction by bFGF and EGF combined with striatal conditioned medium.Partial neuron-like cells with prominence could be found.Immunocytochemieal detection showed that the percentages of NSE and TH positive cells were(72.70±14.81)% and(34.50±15.93)%,respectively.Conclusion BMMSCs can be induced directionally into dopaminergiC neurons by bFGF and EGF combined with striatal conditioned medium in vitro.

Objective To compare the value of conventional ultrasound,hydrosonography and double-contrast enhanced ultrasound in the display capacity of periampullary carcinoma and its relationship with the surrounding tissue.Methods A total of 18 patients with the periampullary carcinoma were diagnosed pathologically or by endoscopic biopsy.Each patient had three imaging modalities by conventional ultrasound,hydrosonography and double-contrast enhanced ultrasound.The display capacity and the relationship between the mass and surrounding tissue were compared with three modalities.Results The display rates of limpid visibilities of carcinoma on conventional ultrasound,hydrosonography and double-contrast enhanced ultrasound were 16.7%(3/18),22.2%(4/18) and 94.4%(17/18),respectively;The display rates of duodenum protrusion on three modalities were 0(0/18),38.9% (7/18) and 72.2% (13/18),respectively,with significant differences among conventional ultrasound,hydrosonography and double-contrast enhanced ultrasound.The double-contrast enhanced ultrasound showed two cases of peripheral vascular invasion and one case of intrahepatic metastasis.Conclusion The double-contrast enhanced ultrasound can increase the display capacity of periampullary carcinoma as a new diagnostic modality for periampullary carcinoma.

OBJECTIVETo study the therapeutic mechanism of Zhizhu Pill (ZP) for treating functional dyspepsia (FD) rats.

METHODSTotally 30 ten-day-old male rats were randomly divided into the normal control group (n =10) and the model group (n = 20). The FD rat model was induced using gastric administration of 0.1% iodoacetamide (IA) combined tail clamping. The model was evaluated when rats were 8-week old. Successfully modeled rats were randomly divided into the model group (n = 10) and the ZP group (n = 10). Rats in the normal group and the model group were administered with normal saline by gastrogavage, while those in the ZP group were administered with ZP Decoction (2 mL/100 g) by gastrogavage. All medication lasted for 7 successive days. The contractile activity in in vitro longitudinal gastric muscle was recorded using Power Lab biological signal collecting system. The expression of growth hormone secretagogue receptor (GHSR) in stomach of FD rats was detected using Western blot and immunohistochemistry (IHC).

RESULTSCompared with the normal group, average frequencies of gastric contraction and changing rates of amplitude obviously decreased in the model group (P < 0.05). Results of Western blot and IHC showed that the expression of GHSR decreased in the model group (P < 0.01). Compared with the model group, average frequencies of gastric contraction and changing rates of amplitude obviously increased in the ZP group (P < 0.05). Results of Western blot and IHC showed that the expression of GHSR increased in the ZP group (P < 0.01).

CONCLUSIONZP could promote the gastric motility in FD rats induced by gastric administration of IA combined tail clamping, and its mechanism might be related to up-regulating GHSR protein level.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Dyspepsia ; drug therapy ; Gastrointestinal Motility ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; metabolism ; Random Allocation ; Rats ; Receptors, Ghrelin ; metabolism