Effects of improved catheter extubation method on urination / 中国实用护理杂志

Objective:To compare the effects between improved catheter extubation method and the traditional one on urination pain, urinary retention, first urination time and first urination volume, and to evaluate the advantage of the improved method.Methods:144 patients with indwelling catheters after operation in our department were randomly divided into observation group and control group, with 72 cases in each group. The control group returned to the ward after surgery and began to clamp the urinary tube to train the bladder function, the catheter was removed by traditional method. Observation group: urination reflex was evaluated before extubation, according to more than 250 ml urine in the urine bag. Pumping the saline of the gas bag and injecting back 0.5ml to keep the wall of the bag smooth, eventually the catheter was excreted when urinating. Urethral pain, urinary retention, first micturition time and first micturition volume of two groups were analyzed.Results:Urethral pain, urinary retention, first micturition time in observation group and control group were 1.47±1.48, (20.44±12.98) min, 95.8% (69/72) and 3.11±1.98, (28.03±27.00) min, 83.3% (60/72), respectively, and the difference between the two groups was statistically significant ( t value was -5.644, -2.148, χ2 value was 6.628, all P<0.05). The first micturition volume in observation group and control group were (258.6±41.57) ml and (248.14±48.82) ml, respectively, and there was no significant difference between the two groups ( t value was 1.377, P>0.05). Conclusion:The improved catheter extubation method could significantly reduce the urethral pain, shorten the time of the first urination, and improve the success rate of self-urination, which of clinical promotion.

Best practice implementation of bedtime and position after diagnostic adult lumbar puncture / 中国实用护理杂志

Objective:By implementing the best practice of bedtime and position after diagnostic adult lumbar puncture,we hope to establish a scientific and standardized nursing routine for lumbar puncture, shorten the bed-rest time after lumbar puncture, and improve the comfort of patients.Methods:By reviewing literatures related to positions after adult lumbar puncture and post-dural puncture headache, six best practice were concluded. By combining the best evidence and the clinical circumstances, the evidenced-based criteria were established and then applied in the Neurology Department.Results:After two rounds of reviews, the results showed that except the 93.3% compliance with the new evidence, all other four criteria had 100% complacence. Comparing before and after applying the evidence, there was no statistically significant difference for the occurrence of post-dural puncture headache or dizziness( P>0.05), there was a statistically significant reduction of back pain from 28.3%(30/106) to 15.1%(18/119)( χ2 value was 5.799, P<0.05) when the evidence was applied. Conclusions:The best practice shows that patients needn′t lie on bed for 4 to 6 hours after lumbar puncture, the occurrence of back pain is lowered and the comfort level of the patient is improved in those who rest with pillow or activities.

Best practice implementation of bedtime and position after diagnostic adult lumbar puncture / 中国实用护理杂志

Objective@#By implementing the best practice of bedtime and position after diagnostic adult lumbar puncture,we hope to establish a scientific and standardized nursing routine for lumbar puncture, shorten the bed-rest time after lumbar puncture, and improve the comfort of patients.@*Methods@#By reviewing literatures related to positions after adult lumbar puncture and post-dural puncture headache, six best practice were concluded. By combining the best evidence and the clinical circumstances, the evidenced-based criteria were established and then applied in the Neurology Department.@*Results@#After two rounds of reviews, the results showed that except the 93.3% compliance with the new evidence, all other four criteria had 100% complacence. Comparing before and after applying the evidence, there was no statistically significant difference for the occurrence of post-dural puncture headache or dizziness(P>0.05), there was a statistically significant reduction of back pain from 28.3%(30/106) to 15.1%(18/119)(χ2 value was 5.799, P<0.05) when the evidence was applied.@*Conclusions@#The best practice shows that patients needn′t lie on bed for 4 to 6 hours after lumbar puncture, the occurrence of back pain is lowered and the comfort level of the patient is improved in those who rest with pillow or activities.

Research progress in the effects of high glucose on signaling pathways in bone metabolism / 中华骨科杂志

Diabetes mellitus (DM),a highly prevalent chronic metabolic disorder,can damage multiple organ systens.For instance,in the skeletal system,DM can lead to osteoporosis,delayed fracture healing or nonunion of fractures,which not only affects the prognosis of bone diseases and quality of life but also leads to huge medical and economic burden.The main reason of these skeletal complications is the disruption of the balance between bone formation and bone resorption,which leads to the disorders of bone metabolism.The mechanism of bone metabolism disorders in DM patients is determined by many biological factors,including high glucose (HG),oxidative stress,accumulation of advanced glycation end products (AGEs),insulin levels,inflammatory factors,growth factors,adipocytokines and hypaglycemic agents.Furthermore,as an important feature of DM,HG has obvious effects on bone metabolism mainly by acting on signal transduction pathways,including PI3K/Akt pathway,cAMP/PKA pathway,wnt pathway,MARK pathway,AMPK/mTOR/ULK1 pathway,BMP pathway,EphrinB2/EphB4 pathway,PPARγ pathway and NF-κB pathway.The above mentioned signal pathways regulate the proliferation,differentiation,apoptosis and aging of cells,such as bone marrow mesenchymal stem cells (BMSCs),osteoblasts and osteoclasts.These research areas have become the current hotspot for investigation.Although some studies has been conducted in these research areas,several limitations are still exist especially in vivo study.Thus,further studies are required.The present article reviews the effects of HG on bone metabolism signaling pathways.The purpose of this review is conducive to further understand the molecular mechanism of bone metabolism regulated by HG,and to provide theoretical basis and orientations for prevention and treatment of diabetic bone diseases.

Effects of sodium butyrate on activity of RAW264.7 cells and osteoclast differentiation / 中华创伤骨科杂志

Objective To investigate the effects of sodium butyrate on the activity of RAW264.7 cells and the osteoclast differentiation.Methods The RAW264.7 cells were treated by sodium butyrate at concentrations of 0,0.25,0.50,1.00,2.00,3.00,4.00 and 5.00 mmol/L,with 3 double pores for each concentration.The cytotoxicity of sodium butyrate on RAW264.7 cells was detected by a CCK-8 kit.The effects of sodium butyrate (0,0.25,0.50 and 1.00 mmol/L) on apoptosis of RAW264.7 cells were detected by Hoechst33342 staining.RAW264.7 cells were induced into osteoclasts by osteoclast differentiation factors.The experiment was carried out in 2 groups (n =3).After induced maturation,the experimental group was treated with 1.00 mmol/L sodium butyrate and the control medium was added only with the same volume of solvent.The number of osteoclasts and the area of bone resorption were observed and compared.The differentiation of RAW264.7 cells was detected by tartrate-resistant acid phosphatase (TRAP) staining.Western blotting was used to detect the effects of sodium butyrate (0,0.25,0.50 and 1.00 mmol/L) on NF-κB-related signaling pathway in RAW264.7 cells.Results Compared with the group of 0 mmol/L sodium butyrate,the activity of cells treated with 1.00,2.00,3.00,4.00 and 5.00 mmol/L sodium butyrate for 24 h was significantly decreased (P ＜ 0.05).Treatment with 1.00 mmol/L sodium butyrate for 24 h induced apoptosis.The number of osteoclasts in the control group and the experimental group were 9.33 ± 2.08 and 4.67 ± 1.16,respectively,showing a significant difference between the 2 groups (t =3.395,P =0.027).The percentages of bone resorption area in the control group and the experimental group were 52.43％ ± 5.38％ and 14.28％ ± 2.72％,respectively,also showing a significant difference between the 2 groups (t =10.970,P ＜ 0.001).Western blot results showed that,compared with other concentrations of sodium butyrate,treatment with 1 mmol/L sodium butyrate on RAW264.7 cells for 24 h led to an increase in the expression levels of cytoplasmic p65,B lymphoma-2 associated X protein and cleaved-caspase 3 and the acetylation of Histone H3 but a decrease in the phosphorylation level of α/β subunit of NF-κB kinase.Conclusions With the increased concentration of sodium butyratecan,the activity of NF-κB may be suppressed and the number of apoptotic cells may increase.1.00 mmol/L sodium butyrate can reduce osteoclast formation and bone resorption area.

Improvement of catalytic activity of Aspergillus terreus lipase by site-directed mutagenesis / 生物工程学报

The catalytic activity of Aspergillus terreus lipase (ATL) was improved by rational design. According to the sequence analysis and homologous modeling, several amino acids involved in the lid domain and substrate binding pocket domains of the acidic lipase ATL were mutated by site-directed mutagenesis, and eight mutants were constructed. These mutants and the wild type lipase ATL were expressed in Pichia pastoris GS115 and the enzymatic properties were characterized. The mutants ATLLid and ATLV218W exhibited higher hydrolytic activity than ATL towards p-nitrophenyl laurate. The kcat values of ATLLid and ATLV218W towards p-nitrophenyl laurate were 39.37- and 50.79-fold higher, and the kcat/Km values were 2.85- and 8.48-fold higher than the wild type, respectively. Although thermostability of these mutants decreased slightly, ATLLid and ATLV218W still exhibited the maximum activity at pH 5.0 and high stability in a broad range of pH (4.0-8.0), which were similar to the wild type. Using homologous modeling and molecular docking technology the mechanism for the improvement of catalytic activity was analyzed. These findings not only shed light on the relationship between the lid domain/substrate binding pocket domain and catalytic activity but also provided comprehensive scheme for further engineering to gain more efficient lipases.

The efficacy of different embolic agents to block bronchial artery for massive hemoptysis secondary to bronchiectasis or pulmonary tuberculosis / 实用医学杂志

Objective To investigate the efficacy of gelatin sponge particles(GSP)or polyvinyl alcohol particles (PVA) for hemoptysis secondary to bronchiectasis or pulmonary tuberculosis. Methods The clinical data on 271 patients with bronchiectasis- or tuberculosis-induced hemoptysis were retrospectively analyzed. The efficacy and rates of recurrence and complications were analyzed. Results A total 271 patients were included in this study, 176 of whom suffered from bronchiectasis and the rest 95 had tuberculosis. One-week cure rate was signifi-cantly higher in bronchiectasis group than in tuberculosis group(73.3%vs. 46.3%,P0.05). No severe complications occurred. Conclusions In-terventional artery embolization therapy for hemoptysis secondary to bronchiectasis is better than tuberculosis-induced hemoptysis,and PVA is more effective than GSP. Recurrence of massive hemoptysis mostly occurrs within one month ,and most of the patients are complicated with blood supply and have a history of hemoptysis.

Identification and characterization of stem cells in an ovarian cancer cell line and examination their drug resistance / 中华妇产科杂志

Objective To isolate side population (SP) cells from an established ovarian cancer (OC)cell line,characterize these cells,and examine their drug resistance. Methods SP and non-SP (NSP) cells were isolated by fluorescence-activated cell sorting (FACS),and cultured in differential conditions,then detected their SP ratio to compare their capability of differentiation and self-renewal. Moreover,SP and NSP cell tumorigenesis was examined by subcutaneous and intraperitoneal injection of these cells to nonobes ediabetic(NOD)-severe combined immundeficient(SCID)mice. Drug resistance to cisplatin was examined by cell counting kit-8 (CCK-8).Results SP cells could be isolated stablly and insistently. There was(4.81 ± 0.43)%of SP cells in the established OC cell line and(4.89 ± 0.33)%of SP cells after cultured the isolated SP cells in differentiation condition,and there was no significant different between these two quantities (P>0.05). However,after cultured the NSP cells,there was only (0.10 ± 0.03)%of SP cells which was significantly lower than that contained in the OC cell line(P<0.01). In the tumorigenesis assay 1.0 × 103 SP cells were injected subcutaneously and formed the xenografted tumors in 6 weeks(3/3),and 1.0×104 NSP cells were injected subcutaneously and did not form xenografted tumors in 12 weeks(0). The tumorigenic capability of SP cells was higher than that of NSP cells(P<0.01). Both the original and the xenografted tumors were low differentiated serous cystadenocarcinomas and expressed the ovarian serous cystadenocarcinomas CA125 marker after stained by HE and immunohistochemistry. Simultaneously,the SP cells were also capable to form tumors as shown by intraperitoneal injection. In the drug resistance assay shown that the 50% inhibitory concentration (IC50) of the SP and NSP cells were respectively(2.33 ± 0.14)μg/ml and(1.60 ± 0.04)μg/ml(P<0.05). After treated the unsorted OC cells with cisplatin,the quantity of SP cells increased to(40.10 ± 4.22)%and there was significant difference,when compared to the untreated cells which was(4.81±0.43)%(P<0.01). The SP cells survival rate was(58.7± 3.3)%when treated with cisplatin at its IC50 dose,and the rate decreased to(7.2±1.3)%(P<0.01)when verapamil was present. Conclusions The SP cells could be isolated from the established OC cell line. They had the capacities of self-renewal,differentiation,and tumorigenesis,and the new tumor demonstrated the original tumor′s phenotype. The SP cells also had stem cells′ biological characteristics and is resistant to cisplatin.

Biomechanical and histological analysis of alveolar distraction osteogenesis in a canine model / 中国组织工程研究

BACKGROUND:Alveolar distraction osteogenesis is an important method for treating alveolar bone atrophy, the osteogenesis process and biomechanics play a crucial role in the fol owing implantation and repair. At present, no related experimental studies are found. OBJECTIVE:To analyze the biomechanical and histological characteristics of alveolar distraction osteogenesis in a canine model. METHODS:Twelve adult mongrel canines received premolars extraction and alveoloplasty in mandible to establish an atrophy alveolar model. After 3 months, a segmental alveolar osteotomy was performed in the randomly selected unilateral atrophy alveolar and two intra-osseous distractors were placed. After a 7-days latency period, the alveolar ridge was augmented at a rate of 1.0 mm/d for 5 days. After a consolidation of 1, 2, and 3 months, the canines were sacrificed and the specimens of the distracted alveolar bone were harvested for clinical, radiographic, histological and biomechanical analysis. RESULTS AND CONCLUSION:The alveolar distractors obtained good healing with surrounding tissue. The atrophy alveolar bones were augmented for (4.80±0.50) mm and (5.12±0.47) mm by clinical and radiographic findings immediately after distraction, respectively. The bone trabeculae in the distracted chamber matured from 1 to 3 months of consolidation by histological analysis. The shearing force of alveolar distraction chamber increased from 1 to 3 months. After 3 months’ consolidation, the shearing force of distracted chamber was comparable to host bone. The histological and biomechanical property of distracted alveolar chamber is comparable to host bone after 3 months’ consolidation.

Enhanced thermostability of Rhizopus chinensis lipase by error-prone PCR / 生物工程学报

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.

Minimally invasive plate osteosynthesis for anterior pelvic ring fractures: a finite element analysis and clinical study / 中华创伤杂志

Objective To investigate the mechanical stability and clinical outcome of minimally invasive plate osteosynthesis of pubic ramus fractures.Methods Stability of minimally invasive plate osteosynthesis and traditional open fixation of pubic ramus fractures was compared in finite element analysis.A retrospective analysis was performed on fractures of pubic rami (126 sides) in 101 consecutive patients treated with minimally invasive plate osteosynthesis from 2005 to 2012.Operation time,intraoperative blood loss and follow-up of fracture healing were evaluated.Results In finite element analysis,traditional open fixation and minimally invasive plate osteosynthesis resulted in the maximum pelvic force of 7.35 MPa and 5.59 MPa,maximum fracture displacement of 4.31 mm and 4.38 mm and relative fracture gap displacement of 0.029 mm and 0.012 mm.Displacement of fracture gap after minimally invasive plate osteosynthesis and traditional open fixation was reduced 26％ and 59％ respectively.In the clinical study,the surgery acquired for pubic ramus fractures averaged 65 minutes with mean blood loss of 94 ml.Follow-up duration was 5-50 months (mean,24.3 months).Reduction of the fracture as assessed using Matta' s criteria was excellent in 118 sides (93.7％),good in eight sides (6.3％).Totally,the fracture was healed within postoperative 12 weeks in 117 sides and within postoperative 6 months in 9 sides.No iatrogenic nerve or vascular injury occurred.Conclusions Minimally invasive plate osteosynthesis is a safe and effective technique for fixation of pubic ramus fractures.Moreover,satisfactory results can be achieved together with less trauma and better cosmetic effect.

Study of the invasiveness and tumour formation of Bcrp1 + HeLa cervical cancer cells / 中华妇产科杂志

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.

Construction of high-quality gene mutant pool in Pichia pastoris by a PCR dependent method / 生物工程学报

We developed a method to construct a gene mutant pool in Pichia pastoris based on in vivo homologous recombination. It was an absolute PCR-dependent method (PDM) and could avoid the disadvantages of traditional mutant pool construction process such as long-experimental period, low pool capacity and inadequate abundance. The method consisted of four steps: (1) construction of recombinant expression plasmid of target gene; (2) design of long primers that have 40-70 bp of homology to expression vector fragments at both ends and amplification of target gene by error-prone PCR, DNA Shuffling or other methods; (3) PCR amplification of expression vectors fragments; (4) mixture of gene and vectors by appropriate mole ratio, electroporation, formation of expression cassette in vivo, homologous recombination with host genome and achievement of mutant pool. Screening from this library, we obtained mutants with improved enzyme activity, protein expression level and thermostability. In conclusion, PDM was very efficient and convenient with advantages of shortened pool construction cycle from 2 weeks to 3 days, enlarged pool capacity from the original 10(3)-10(4) to more than 10(5), with a positive rate of more than 95%.

Prevalence of Three Shrimp Viruses in Zhejiang Province in 2008 / 中国病毒学·英文版

White spot syndrome virus(WSSV), Taura syndrome virus(TSV)and Infectious hypodermal and haematopoietic necrosis virus(IHHNV)are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. Shrimps samples were collected from 12 farms in Zhejiang province, China, in 2008 and analyzed by PCR to determine the prevalence of these viruses. From the 12 sampling locations, 8 farms were positive for WSSV, 8 for IHHNV and 6 for both WSSV and IHHNV. An average percentage of 57.4% of shrimp individuals were infected with WSSV, while 49.2% were infected with IHHNV. A high prevalence of co-infection with WSSV and IHHNV among samples was detected from the following samples: Bingjiang(93.3%), liuao(66.7%), Jianshan(46.7%)and Xianxiang(46.7%). No samples exhibited evidence of infection with TSV in collected samples. This study provides comprehensive information of the prevalence of three shrimp viruses in Zhejiang and may be helpful for disease prevention control in this region.

Research of the characterization of Bcrp1+ HeLa cells / 中华妇产科杂志

Objective To make sure whether Bcrp1 is the marker of cervical cancer stem-like cells or not by studying the characterization of Bcrp1+ HeLa cells.Methods Immunofluorescence stained flow cytometry and electron microacope were used to sort and observe uhrastructures of Bctp1+ and Bcrp1- HeLa cells.Flow cytometry wag used to identify the cycle and the rate of apoptosis with annexin V in two group cells.The expression of proliferating cell nuclear antigen (PCNA)and caspase-3 were tested using western blot methed.Results (1)There were 7.1% Bcrp1+ cells and 92.9% Bcrol- cells in HeLa cells.Bcrp1+ HeLa cells were large in size of nuclear and nucleoli are clear.and there were rich of cytomicrosome and rough endoplasmic reticulum.After sorted and cultured for 24,48,72 hours,the adhesion in Bcrp1+ cells were 72.8%,81.1%,80.4%,respectively.While,they were 3.3%,18.7%,12.6%at each time for Bcrpl- cells(all P<0. 05 ). (2) There are more S phase cells in Bcrp1+ cells than that in Bcrp1- cells (54. 1% vs 21.1%, P <0. 05) ,while the percentage of G0/G1 and G2/M in Bcrp1 - cells were highter than those in Bcrp1 + cells (53.0% vs 44. 4% ,25.9% vs 1.5% ; all P <0. 05 ). The rate of apoptosis in Bcrp1+ cells was lower than that in Bcrp1 - cells (0. 2% vs 5.3%, P < 0. 05 ). ( 3 ) The expression of PCNA in Bcrp1 + cells was higher than that in Bcrp1- cells (3140 vs 2255, P< 0. 05 ), while the expression of caspase-3 of Bcrpl + cells was lower than that in Bcrp1 - cells ( 1970 vs 3551, P < 0. 05 ). Conclusion There are more vigor and ability of proliferation and lower rate of apeptosis in Bcrp1 + HeLa cells than those in Bcrp1 - cells ,which may be some characters of cervical cancer stem cells.

Increasing activity of Rhizopus chinensis CCTCC M201021 lipase by directed evolution-error prone PCR / 生物工程学报

Directed evolution strategy (error-prone PCR) was conducted to improve the activity of lipase from Rhizopus chinensis CCTCC M201021. Through two rounds of ep-PCR and pNPP top agar screening, two optimum mutant strains 1-11 and 2-28 were obtained with 2 and 4 fold of enzyme activity higher than that of parent strain, respectively. DNA sequencing of mutant lipase 2-28 revealed four amino acid substitutions: A129S, K161R, A230T, K322R. According to the simulated protein structure of Rhizopus chinensis lipase, A129S, K161R, A230T were located on the surface of the protein. A230T substitution improved the stability of the alpha-helix loop. K322R, near the catalytic center of lipase, located at a loop, formed a salt-bridge with a nearby aspartic acid (negative charged). Electrostatic force pulled the loop to the opposite direction of the substrate channel and made it easier for substrate to enter the lipase catalytic domain. Purified lipase was characterized and the result showed that Km of 2-28 lipase decreased by 10% compared with Km of the parent lipase, and Kcat was 2.75 fold improved than that of the original lipase.

Isolation and detection of CK17~+,CD44v5~+ and Bcrp1~+ cells in HeLa cell line of cervical carinoma / 吉林大学学报(医学版)

Objective To identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells,and investigate the relationship between them and select cervical cancer stem cell markers.Methods Flow cytometry was used to identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells;after isolation,the expressions of CK17 and CD44v5 in Bcrp1+ and Bcrp1-phenotype HeLa cells were measured by immunocytochemistry.Results HeLa cells contained 11% Bcrp1+ cells,0.7% CK17+ cells and 0.1%CD44v5+ cells.Bcrp1+ HeLa cells contained CK17+ and CD44v5+ HeLa cells,but the Bcrp1-Hela cells did not(P

Professor JIANG Yu-ren’s clinical experience in treating measles / 中华中医药杂志

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.

Primary determination for activity and expression of Stx 2a’-LHRH chimeric toxin / 中国病理生理杂志

AIM: To express chimeric toxin Stx2a’-LHRH a nd to investigate the cytotoxic activity of recombinant toxin Stx2a’-LHRH to huma n carcinoma cells.METHODS: Stx2a’-LHRH sequences that added the res tri ction endonucleases NcoⅠ and EcoRⅠ at the 5' and 3 ends were amplified by PCR a nd digested with appropriate restriction enzymes. The digested fragment was subc loned into the vector obtatined by digestion of plasmid pET-28a(+) with NcoⅠ an d EcoRⅠ. E. coli BL21 (DE3) cells were transformed with plasmids of interst and cultured in LB medium containing ampicillin. Expression of the recombinant protein was induced by the addition of isopropylthio-?-D-galactoside (IPTG). T h e cytotoxity of Stx2a’-LHRH to Hep-2 cells was observed under the microscop y. RESULTS: Recombitant plasmid pET-SL was constructed successfu lly and the clones expressing pET-SL stablely were obtained. A special electroph oretic band in SDS-PAGE (a glycoprotein of 28kD) was noted. Stx2a’-LHRH killed He lp-2 cells clearly. CONCLUSION: In this study, construction of c himeric toxin Stx2a’-LHRH and its expression were described. Moreover, it has o bvious cytotoxity to Hep-2 cell. These finding could open up new vistas in the s tudy of targeted durgs.