Objective To construct mycobacterial membrane-anchored expression vector and to analyze expression level and sub-cellualr localization of exogenous target protein. Methods Based on the mycobacterial intracellular expression vector pMFA42 which contained a strong promoter of pfurAma mutant, the signal sequence of Mycobacterium tuberculosis(Mtb) 19×103 lipoprotein (19SS) was synthesized and was then cloned into the downstream of pfurAma mutant to generate the mycobacterial membrane-anchored expression vector pMFA42M. The coding gene of enhanced green fluorescent protein(EGFP) was amplified by PCR, and then sub-cloned into these two vectors described above to construct recombinant EGFP fused and membrane-anchored strains, respectively. The coding genes of Mtb immuno-dominant antigens Ag85A and its chimera Ag856A2 were then sub-cloned intothe membrane-anchored construct pMFA42MG to produce recombinant Mtb antigen EGFP fused-expression strains. After that, expression levels and sub-cellualr localization of exogenous target protein were further analyzed by Western blot and flow cytometry sorting(FCS), and the fluorescence intensities of recombinant EGFP- expressed strains were observed in vitro directly and after transfection of murine macrophage cell line RAW264.7. Results The novel mycobacterial membrane-anchored expression vector was constructed successfully by introduction of signal sequence of Mtb 19×103 lipoprotein. Using of EGFP as model antigen, exogenous target protein was demonstrated to be expressed with high level and could be anchored into cell membrane of recombinant mycobaterial strains. Conclusion A novel mycobacterial membrane-anchored expression vector was constructed successfully to research recombinant BCG and functions of mycobacterial membrane proteins, and the constructed EGFP-expressed recombinant strains could also be used to research cytophagy in cell model and mycobacterial colony and translocation in animal immunization as model indicator bacteria.

Objective: To investigate the antihypertensive time-effect and dose-effect features of Sancao jiangya decoction(SCD).Methods: The blood pressure of spontaneously hypertensive rats at different time points were measured after treatment with Sancao jiangya decoction of low,middle,high concentration by tailartery blood pressure measurement for conscious rats.Results: The blood pressure was decreased at 2 hours after drug taken,there were significant dose-effect relationship between the antihypertensive effect and the low,middle,high dose.At 4h after drug taken,the high,middle dose had dose-effect correlation,but the low-dose had no antihypertensive effect.Further research on the middledose shows that the blood pressure reduced at 1h after drug taken,and the stable antihypertensive effect was keeping during 1-4h,the blood pressure began to rise at 6h,and got back to the level before drug taken at 8h.Conclusion: To choose the Middle-dose(10.4g crude drug/kg body weight) and 2h after drug taken is appropriate for SCD's use.This result laid a substantial foundation for further research on effects evaluation and mechanism of antihypertensive medicine.

Compatibility mechanism study of Chinese herbal compound (CHC) has been one of key contents in Chinese medical research, but the present research methods are not suitable for its own features due to its complexity, which has restricted the process of modernization and intemationalization of Chinese medicine. In this paper, we addressed that the compatibility is closely correlated to their in vivo metabolic processes. The preparation of active small molecules monoclonal antibodies in herbs and testing a variety of effective compositions simultaneously using immunoassay can clarify the in vivo metabolism and mutual interactions of Chinese herbs, which is a new thought of studying the compound compatibility mechanisms.
Antibodies, Monoclonal ; administration & dosage ; Drug Combinations ; Drug Incompatibility ; Drugs, Chinese Herbal ; administration & dosage

Laser ablation inductively coupled plasma mass spectrometry ( LA_ICP_MS ) was applied for the determination of doping element chromium( Cr) content and distribution in Cr∶ZnSe crystals. Several different Cr∶ZnSe crystals were prepared by diffusion method as reference material to solve the problem of accurate quantization. The homogeneity of Cr in these samples was characterized by LA_ICP_MS and the concentrations achieved by inductively coupled plasma atomic emission spectrometry ( ICP_AES ) . With signal pot and line scan sampling, the present method provided effective position and content distribution information of Cr in ZnSe crystals, achieved the in situ analysis. The correlation coefficient of Cr in calibration curve was 0. 9992 and the detection limit was 0. 08 mg/kg. It could provide effective means for the distribution statistics of doping element in different growth condition crystals.

Objective To study the application of multi-slice spiral CT(MSCT) portography in diagnonsis of pancreatic portal hypertension. Methods Forty-seven patients with lesion in body or tail of pancrease,47 normal subjects and 126 patients with portal hypertension underwent MSCT portography with LightSpeed 16 CT scanner. The inner diameter of portal system and the main collateral veins were measured in maximun intensity projection (MIP) image. The volume of liver and spleen were also measured in volume rendering (VR) image. The liver parenchyma and main portal vein enhancement in portal vein phase were also taken. The endoscopy examination was made in 57 patiens with portal hypertension. Results In 47 patiens with lesion in body or tail of pancreas , stenosis or occlusion of spleen vein were found in 38 patients(pancreatic portal hypertension in 27 patients, chronic and acute pancreatitis in 11 patients). In 38 patients with pancreatic portal hypertension, esophageal varices was found in 5 patients(13.2%), gastric fundus varix in 25 patients (65.8%), gastric body variees in 22 patients (57.9%), short-gastric vein/post-gastric vein(SGV/PGV) in 26 patients (68.4%), coronal gastric vein in 26 patients (68.4%),dilated gasto-omenta vein in 24 patients(63.2%), mesenterica varicesin 1 patient, splenic vein occlusion in 14 patients (36.8%), splenic vein stenosis in 23 patinets(63.2%). Conclusions The patients with pancreatic portal hypertension were demonstrate characteristic changes in MSCT portography. The MSCT portography is helpful in etiological diagnosis of pancreatic portal hypertension by supplying images in vessel morphology.

Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.
Animals ; Antibodies ; analysis ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Glucosides ; chemistry ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; chemistry ; immunology ; Serum Albumin, Bovine ; chemistry ; immunology

Maple syrup urine disease is a common amino acids metabolic disease. In most patients, onset occurs in the neonatal period and infancy. In this study, the case of a school boy with acute encephalopathy due to late-onset maple syrup urine disease is summarized. The boy (8.5 years) was admitted because of acute encephalopathy after suffering from infection for two days at the age of eight and a half years. Metabolic acidosis, hyperuricemia and decreased protein level in cerebrospinal fluid were found by general laboratory tests. Magnetic resonance imaging of the brain revealed signal intensity abnormalities in the bilateral cerebellum dentate nucleus, brainstem, thalamus, putamen, caudate nucleus and cortex of the cerebral hemispheres. On T1WI and T2WI scanning, hyperintensive signal was found. Blood leucine and valine were significantly elevated. Urinary 2-hydroxy isovaleric acid, 3-hydroxybutyric acid, 2-keto isovaleric acid, and 2-keto acid also increased. Both the blood amino acid and urine organic acid profiles led to the diagnosis of maple syrup urine disease. In the acute period, the patient was treated with a large dose of vitamin B1, glucose, L-carnitine and a protein-restrict diet. The patient's condition improved significantly after five days of treatment, and he recovered completely two days later. Afterwards, treatment with vitamin B1, L-carnitine and a protein-restrict diet (1 g/kg/day) was continued. One and a half months later, blood amino acids and urine organic acids returned to normal. Magnetic resonance imaging of the brain also indicated a great improvement. It was concluded that inborn metabolic disease should be considered in the patients with an onset similar to acute encephalopathy. Early diagnosis and proper treatment can prevent brain damage and improve prognosis.
Acute Disease ; Brain Diseases ; etiology ; Child ; Humans ; Magnetic Resonance Imaging ; Male ; Maple Syrup Urine Disease ; complications ; diagnosis ; therapy

OBJECTIVETo explore the technique and therapeutic effect of bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral shaft fractures in young children. Methods:From June 2006 to June 2012,45 young children with femoral shaft fractures were treated by bridge wire splint fixation with ankle dorsiflexion,which was designed according to arch bridge mechanical principle and structure. There were 31 males and 14 females with an average age of 3.2 years old ranging from 8 months to 5.5 years old; 14 cases were upper 1/3 femoral fractures,26 cases were middle 1/3 femoral fractures,5 cases were lower 1/3 femoral fractures; 20 cases were transverse fractures, 14 cases were oblique fractures,6 cases were spiral frac- tures, and 5 cases were comminuted fractures. X-ray, follow-up imaging changes,clinical curative effect and complications were assessed.

RESULTSForty-five patients were followed up for 6 to 21 months (averaged 12 months). All fractures were reached clinical bone healing after 5 to 7 weeks (averaged 6 weeks) fixation. Seven cases appearred limb soft tissue complications, including buttocks bedsore,dorsal foot and Achilles tendon epidermal necrosis, and healed after dressing and removal of external fixation. During follow-up,the original overlap angle and lateral displacement were remodeled, and limbs were restored to the normal line of force and bone structure. According to Flynn standard, 35 cases got excellent results, 8 cases good, 2 cases fair.

CONCLUSIONThe bridge wire splint fixation with ankle dorsiflexion for the treatment of femoral. shaft fractures in young children (less than 6 years old) is safe,feasible, simple,and has raliable effect, which can be applied in primary hospitals.

Ankle ; surgery ; Bone Wires ; Child ; Child, Preschool ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; instrumentation ; Humans ; Infant ; Male ; Treatment Outcome

OBJECTIVETo assess the therapeutic effect and mechanism of electroacupuncture combined with auricular point tapping and pressing on the obese women with polycystic ovary syndrome.

METHODSThirty-nine cases of obese women with polycystic ovary syndrome were treated with electroacupuncture combined with auricular point tapping and pressing, body points as Tianshu (ST 25), Fenglong (ST 40), Guanyuan (CV 4) and Siman (KI 14) etc. were selected, and ear points as Kou (mouth), Wei (stomach) and Pi (spleen) etc. were selected. After 3 courses, the therapeutic effect, the body mass index (BMI), the waist circumference (WC) and the changes of the serum insulin (Ins) and testosterone (T) were compared before and after treatment.

RESULTSOf the 39 cases, 10 cases were cured, 25 cases were effective, 4 cases were ineffective, with a total effective rate of 89.7%; there were significant differences in BMI, WC, Ins and T of the patients compared with that before treatment (all P < 0.01).

CONCLUSIONElectroacupuncture combined with auricular point tapping and pressing has a good clinical effect on obese women with polycystic ovary syndrome, the treatment mechanism may realized by regulating the serum insulin and the testosterone of the patients.

Acupuncture Points ; Adult ; Ear Auricle ; pathology ; Electroacupuncture ; methods ; Female ; Humans ; Insulin ; blood ; Middle Aged ; Obesity ; complications ; Polycystic Ovary Syndrome ; blood ; complications ; therapy ; Testosterone ; blood ; Treatment Outcome ; Young Adult

The relationship between tyrosine phosphorylation (TP) and protein expression of insulin receptor (InsR) and insulin resistance (IR) in patients with gestational diabetes mellitus (GDM) was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM (GDM group, n=22), normal pregnant women (normal pregnancy group, n=22) and normal non-pregnant women (normal non-pregnant group, n=13). Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidase assay and immunoradioassay. The results showed that the levels of FPG (5.61±0.78 mmol/L), FINS (15.42±5.13 mU/L) and Homeostasis model assessment-IR (HOMA-IR) (1.21±0.52) in GDM group were significantly higher than those in normal pregnancy group (4.43±0.46 mmol/L, 10.56±3.07 mU/L and 0.80±0.31 respectively) (P<0.01). The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in normal non-pregnant group (7.56±2.31 mU/L and 0.47±0.26 respectively) (P<0.01). There was no significant difference in the InsR expression level among the three groups (P>0.05). TP of InsR with insulin stimulation was significantly decreased in GDM group (0.20±0.05) as compared with normal pregnancy group (0.26±0.06) (P<0.01). TP of InsR with insulin stimulation in normal pregnancy group was lower than that in normal non-pregnant group (0.31±0.06) (P<0.01). TP of InsR with insulin stimulation was negatively related with HOMA-IR in GDM group (r=-0.525, P<0.01). There was no correlation between the protein expression of InsR and HOMA-IR in GDM group (r=-0.236, P>0.05). It was suggested that there is no significant correlation between the protein expression of InsR in skeletal muscle and IR in GDM, but changes in TP of InsR are associated with IR in GDM.