Professor JIANG Yu-ren,a famous pediatrist of TCM in present age,has been engaged in clinic,teaching and scientifi c research of TCM pediatrics for 60 years.His achievement is remarkable.Especially,he has rich clinical experience in acute febricity of paediatrics,such as critical encephalitis B and serious measles complicated by pneumonia.He puts forward the rule of treating measles with favourable prognosis by method of dispersing-promoting,cooling-relieving and nourishing ying,treating measles with reverse prognosis by method of clearing heat,cooling blood and reviving yang and treating measles complicated by pneumonia should use the method of promoting pathogenic factors,removing toxic substance and relieving exhaustion.

Directed evolution was conducted to improve the thermostability of lipase from Rhizopus chinensis CCTCC M201021. Mutations were introduced by two rounds of error-prone PCR and mutant lipase was selected by fast-blue RR top agar screening. Two positive variants were selected in the first-round and four in the second-round screening process. Ep2-4 was proved as the most thermostable lipase and its DNA sequencing revealed three amino acid substitutions: A129S, P168L and V329A. Compared with the parent, its half-life at 60 degrees C was 5.4- times longer and T50 was 7.8 degrees higher. Purified lipase of Ep2-4 was characterized and the result shows that its thermostability improved without compromising enzyme activity. According to the mimicked protein structure, mutation A129S formed a hydrogen bond with Gln133 and improved the thermostability by increasing the hydrophilicity and polarity of protein; mutation P168L by forming a hydrophobic bond with the nearby Leu164.
Cloning, Molecular ; Directed Molecular Evolution ; methods ; Enzyme Stability ; genetics ; Hot Temperature ; Industrial Microbiology ; Lipase ; chemistry ; genetics ; Mutation ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Protein Engineering ; methods ; Rhizopus ; enzymology

Objective To make sure whether or not Bcrp1 is the marker of cervical cancer stem cells or not by studying the invasive ability and formation of tumors of Bcrp1 + phenotype HeLa cells.Methods The tumor cell migration and invasion assay were used by boyden chamber to identify the invasive ability of Bcrp1 + phenotype HeLa cells.The formation of tumors in vivo experiments were completed,in which the two groups of cells with different concentrations were inoculated in non obese diabetes-severe combined immunodeficiency disease ( NOD/SCID ) mice ( 1 × 104,1 × 105,1 × 106/ml ) and the differences of time,rate and volume in the formation of tumors between two groups were observed.Results ( 1 ) In the invasion assay,the amount of cells that invaded through the artificial basement membrane in Bcrp1 + group were 99 ± 14,which was significantly greater than those in Bcrp1- group ( 57 ± 13,P ＜ 0.05 ) ; the length of the Bcrp1 + group was ( 366 ± 52 ) μm,which was significantly greater than the Bcrp1 - group ( 301 ± 54) μm ( P ＜ 0.05 ).( 2 ) Following transplantation of 1 × 104 cells,only the Bcrp1 + cells formed tumors in NOD/SCID mice.When 1 × 105 or 1 × 106 cells were transplanted,the tumor incidence and the tumor mass were greater in the Bcrp1 + groups than those in the Bcrp1 - groups ( P ＜ 0.05 ).Conclusion Bcrp1 + HeLa cell have the greater capacity of invasive and the tumorigenicity,which may contain cancer stem cells.

Objective To compare the clinical effect of needle knife therapy and traction for treatment of cervical spondylosis .Methods 156 patients with nerve root type of cervical spondylosis patients definitely diag -nosed,randomly divided into two groups ,the treatment group of 78 cases,with small needle knife therapy;the control group of 78 cases,the cervical traction therapy ,combined with traditional Chinese medicine .The clinical efficacy was compared between the two groups .Results The treatment group 70 cases were cured(89.6%),the total efficiency of 96.9%;control group 44 cases were cured(56.7%),the total efficiency of 83.3%.The cure rate and total effective rate was significantly better than the control group (χ2 =22.02,11.57,all P<0.05).Conclusion The needle knife therapy to treat cervical vertebra disease is better than that of cervical traction ,has good clinical curative effect .

Patients with primary liver cancer are always associated with hypersplenism .The traditional treatment of hypersplenism is splenectomy .With the development of interventional imaging ,partial splenic emboli-zation has been used extensively in clinical practice .Hepatocellular carcinoma with hypersplenism patients by par-tial splenic embolization can obviously improve the blood , reduce the complications of portal hypertension .PSE combined with other treatment can prolong the survival of patients with hepatocellular carcinoma .In this paper ,the clinical application of PSE is reviewed .

Objective To set up the quality standard of Shenbai concentrated pills. Methods Radix Astragali, Radix Paeoniae Rubra and Rhizoma Imperatea in Shenbai concentrated pills were identified with the method of TLC. The concentration of salvianolic acid B was determined with the method of HPLC. Results For TLC, the chromatogram spots of Radix Astragali, Radix Paeoniae Rubra and Rhizoma Imperatea were well separated and without interference in their negative controls. Quantitative analysis of HPLC showed that the average recovery of salvianolic acid B was 99.74% (RSD=2.08%) with the RSD of precision, reproducibility and stability was 1.74%, 1.19% and 1.70%, respectively. Conclusions The methods set up by this study are easy, accurate and stable, and can be used as the quality control standard of Shenbai concentrated pills.

Objective:To explore the effect of radiotherapy on the FHIT protein expression and cell apoptosis of cervical squamous carcinoma and discuss the relationship between FHIT protein expression and cell apoptosis. Methods:Expression of FHIT protein was measured by immunohistochemical method and cell apoptosis was detected by TdT-mediated dUTP terminal nick end labeling (TUNEL) staining in 50 cases of squamous cell cervical carcinoma at ⅡB-ⅢB stages before, during (Dt 10 Gy and Dt 30 Gy), and after radiotherapy. Results:Of the 50 patients, the positive rates of the expression of FHIT protein was 56% at Dt 10 Gy, 68% at Dt 30 Gy, and 84% after radiotherapy, which were significantly increased compared with that before radiotherapy (36%, P<0.05). The positive rates of cell apoptosis was 52% at Dt 10 Gy, 64% at Dt 30 Gy and 78% after radiotherapy, which were significantly elevated compared with that before radiotherapy (28%, P<0.05). In the process of radiotherapy, cell apoptosis was positively related to the expression of FHIT protein (P<0.05). Conclusion:Radiotherapy reinforces the expression of FHIT protein and induces apoptosis cocurrently. FHIT protein has regulatory effects in cell apoptosis induced by radiotherapy.

Objective To study the effects of total saponins of Chaenomeles speciosa on release of β-hexosaminidase from rat basophilic leukemia-2H3 ( RBL-2H3 ) mast cells. Methods After rat RBL-2H3 mast cells were prepared, total saponins of Chaenomeles speciosa and the RBL-2H3 mast cells were co-cultured.The toxic effects of total saponins of Chaenomeles speciosa on mast cells were detected by MTT method, β-hexosaminidase release rate was measured by fluorescence quantitative spectrophotometric method, and cell supernatants of tumor necrosis factor-α( TNF-α) release were detected by ELISA. Results After total saponins of Chaenomeles speciosa were co-cultured with RBL-2H3 mast cells with different antigen stimulation, β-hexosaminidase release rates and the levels of TNF-α of mast cells significantly decreased compared with the control group. Conclusion Total saponins of Chaenomeles speciosa inhibit the degranulation of RBL-2H3 mast cells in a dose dependent manner, which provids basis for studying mechanism of inhibiting allergic reactions.

BACKGROUND:For unstable intertrochanteric fracture repair, there are two views:extramedul ary fixation or intramedul ary fixation. Theoretical y, intramedul ary fixation is in line with the principles of minimal y invasive, more mechanical and biological advantages. However, evidence-based medicine and related studies have shown that compared with the extramedul ary fixation, intramedul ary fixation did not reflect the proper theoretical advantage. OBJECTIVE:To compare the biomechanical performance of two kinds of internal fixation systems:dynamic hip screw and Gamma nail which commonly used in repair of intertrochanteric fractures by finite element method, and to evaluate the advantages and disadvantages of dynamic hip screw and Gamma nail in treatment of unstable intertrochanteric fracture. METHODS:Three-dimensional finite element model of human femur unstable intertrochanteric fractures (31-A2;AO fracture classification), and the three-dimensional finite element models of dynamic hip screw and Gamma nail were established respectively, and were fixed according to the requirement of orthopedic surgery. The reference load which the joint bearing was at the peak time in adult step state period with the body mass of 700 N was stimulated. The stress distribution of bone, bone-internal fixation model, nail or screw, the strain and deformation of fracture location on the surface of the bone and bone-internal fixation model, the stress distribution along the femur and the loading transfer condition along the internal fixator and the like were analyzed. RESULTS AND CONCLUSION:Dynamic hip screw and Gamma nail have good sliding compression features which could make the continuous and dynamical y axial compression of the fractured section. Under the load conditions, the displacement value of dynamic hip screw was larger. In the treatment of unstable intertrochanteric fracture, Gamma nail was stronger than dynamic hip screw. Two kinds of internal fixations al make the bearing load of the proximal femur reduce. In the treatment of 31-A2 type unstable intertrochanteric fractures, we should choose dynamic hip screw for fixation if the femoral calcar was not seriously crushed, little defect or no defects and can immediately rebuild the stability of femoral calcar during operation, otherwise, stronger internal fixation, namely Gamma nail fixation should be chosen.

BACKGROUND:Nanosilver has significant antibacterial properties, and nanohydroxyapatite has good biological activity and mechanical strength, while their mixture cannot only promote bone formation but also have antibacterial properties. OBJECTIVE:To observe the effect of nanosilver and nanohydroxyapatite mixed filing on the osseointegration of immediate implants. METHODS: Nine New Zealand white rabbits were randomly divided into experimental group (n=6) and control group (n=3) after four incisors from the upper and lower jaw were extracted. The mixture of nanosilver and nanohydroxyapatite was filed into the tooth socket in the experimental group, while nanohydroxyapatite was filed into the tooth socket in the control group. Titanium screw was immediately implanted into both groups. The intact maxila and mandibular specimens were harvested at the 4th, 8th, and 12th weeks after operation. Gross observation, X-ray bone density analysis, torque test and histological observation were conducted. RESULTS AND CONCLUSION:The gray value and maximum torque value of regenerated osseous tissue at different time points in experimental group were significantly higher than those in the control group (P < 0.05). Within 12 weeks of implantation, the bone formation rate and maturity of new bone tissue were higher in the experimental group compared with the control group, and no inflammatory cel infiltration occurred. At the 4th week after implantation, there was a large amount of inflammatory cel infiltration, and few inflammatory existed at the 8th week after implantation. These results demonstrate that compared with nanohydroxyapatite alone, the mixture of nanosilver and nanohydroxyapatite shows better antibacterial effect, biocompatibility and osteoinductive ability, which may accelerate osseointegration and promote osteogenesis.