Clinical outcomes of reverse shoulder arthroplasty for the treatment of failed fixation of proximal humeral fracrtures in the elderly patients / 中国骨伤

OBJECTIVE@#To evaluate the clinical outcomes of reverse total shoulder arthroplasty as a revision procedure for the failed fixation of proximal humeral fractures in the elderly patients.@*METHODS@#A retrospective analysis was performed on 8 patients with failed internal fixation of proximal humeral fractures from May 2014 to March 2020, including 3 males and 5 females, aged from 65 to 75 years old. All 8 patients underwent reverse total shoulder arthroplasty, and the mean time between initial fixation and reverse total shoulder arthroplasty ranged from 8 to 16 months. Range of motion(ROM), University of California at Los Angeles(UCLA) shoulder score, visual analogue scale (VAS), self-rating anxiety scale(SAS), and Constant-Murley score of shoulder function were assessed pre-operatively and at the last follow-up. Complications relating to the surgery were recorded.@*RESULTS@#All 8 patients successfully followed up. The mean follow-up after reverse total shoulder arhroplasty ranged from 16 to 28 months. The range of motion (forward flexion, external rotation, abduction and internal rotation) of the affected shoulder was significantly improved after surgery, and the post-operative VAS, SAS and UCLA scores were also significantly improved. For the Constant-Murley score of shoulder joint function, the total scores and the subscores of pain, daily activities, range of motion and strength test at the last follow-up were all significantly improved. Scapular glenoid notch was observed in patient, which was evaluated as grade 1 on imaging. All the other patients did not develop specific or non-specific complications.@*CONCLUSION@#Reverse total shoulder arhroplasty is an appropriate treatment as a revision surgery for failed fixation of proximal humeral fractures. It has shown satisfactory clinical outcomes, accelerating the rehabilitation of shoulder function and improving the quality of life.

Factors Related to Tympanic Membrane Perforation in Children with Acute Suppurative Otitis Media / 中国医学科学院学报

Objective To explore the factors related to tympanic membrane perforation in children with acute suppurative otitis media,and to provide reference for clinical practice. Methods We reviewed the clinical data of 1274 children with acute suppurative otitis media from February 2017 to May 2020,and analyzed the factors related to tympanic membrane perforation. Results Tympanic membrane perforation occurred in 67 out of the 1274 children with acute suppurative otitis media,with the incidence of 5.27%.The univariate analysis showed that 11 factors including the duration of onset(

A guide to human microbiome research: study design, sample collection, and bioinformatics analysis / 中华医学杂志(英文版)

The purpose of this review is to provide medical researchers, especially those without a bioinformatics background, with an easy-to-understand summary of the concepts and technologies used in microbiome research. First, we define primary concepts such as microbiota, microbiome, and metagenome. Then, we discuss study design schemes, the methods of sample size calculation, and the methods for improving the reliability of research. We emphasize the importance of negative and positive controls in this section. Next, we discuss statistical analysis methods used in microbiome research, focusing on problems with multiple comparisons and ways to compare β-diversity between groups. Finally, we provide step-by-step pipelines for bioinformatics analysis. In summary, the meticulous study design is a key step to obtaining meaningful results, and appropriate statistical methods are important for accurate interpretation of microbiome data. The step-by-step pipelines provide researchers with insights into newly developed bioinformatics analysis methods.

Elimination of a disulfide bond in the light chain of coagulation factor VIII improves secretion of a BDD-FVIII variant with an engineered inter-chain disulfide / 药学学报

The coagulation VIII factor (FVIII) contains eight pairs of disulfide bonds, which are involved in maintaining its structure and function. It has been demonstrated that the disulfide bond between Cys1899/Cys1903 of the A3 domain in the light chain impedes secretion. In our previous work, an engineered inter-chain disulfide in the B domain-deleted FVIII (BDD-FVIII) promoted heterodimer assembly and secretion of separately expressed heavy and light chains. In this study, we constructed two BDD-FVIII variants, one of which contains an engineered inter-chain disulfide bond (F8C) between Met662 > Cys and Asp1828 > Cys mutations and another contains an endogenous A3 domain with a disrupted disulfide bond from F8C (F8CG) by replacement of Cys1899 and Cys1903 with Gly in F8C. We explored their function and secretion. By transducing F8C and F8CG into HEK293 and COS-7 cells, the formation of disulfide bonds and the secretion and coagulation activity of the two variants in the culture media and their binding affinity for von Willebrand factor (vWF) could be observed. The results show that variants F8C and F8CG are mainly the disulfide bonded heavy and light chain dimer, while the wild type BDD-FVIII (F8) is dominated by the easily dissociated heavy and light chain dimer. The secretion and activity of F8C was significantly higher than that of F8, while the secretion and activity of F8CG was significantly higher than that of F8C. The vWF binding of the two variants is similar to F8. This indicates that the BDD-FVIII variant F8CG may be attractive molecule for protein replacement and as a transgene in gene-therapy strategies. These findings are encouraging for future studies targeting disulfide bond elimination for further enhancement of FVIII secretion.

Status of Health Resource Allocation of Medical Institutions in Poverty-stricken Areas / 昆明医科大学学报

Objective To understand the status quo of health resource allocation of medical institutions in poverty-stricken areas, and to provide decision-making basis for rationalizing health and poverty alleviation policies and improving the overall service capacity of medical institutions in poverty-stricken areas. Me thods The overall institutions, bed capacity and staffs in medical institutions in 680 poor counties were analyzed. Re s ults The proportion of government health expenditure in 14 concentrated areas was lower than 15％. The largest number of health institutions was 349 in the Dabie Mountains and 70 in Tibet, and the number of beds was lower than the national average level of 5.11.The largest number of health technical staff for 1 000 people of the four provinces is 4.42 people, the smallest number is 2.72 in Wumeng mountain area;the registered nurses (number) for 1 000 people is up to 1.56 people in the Luo Xia mountain area, the lowest Tibet, only 0.39 people. Thousands of population practice (assistant) physician number of Tibetans is up to 2.98 people, the lowest is 1.07 for the Xinjiang Southern Xinjiang three states; health care than the lowest in Tibet 1:0.54. Conclus ion At present, China's centralized contiguous poverty-stricken areas of county-level medical institutions is extremely short of resources, and the health resource allocation is uneven.

Application of drilling columnar autogenous iliac bone graft and clinical analysis of postoperative complications in the donor bone region / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical effect of drilling columnar autogenous iliac bone graft and analyze the effect of bone grafting on postoperative complications in donor site.</p><p><b>METHODS</b>From March 2014 to October 2016, 68 patients with autogenous iliac bone graft were retrospectively analyzed, and divided into drilling group and osteotomy group, 34 patients in each group. In drilling group, there were 24 males and 10 females with an average age of (40.06±5.60) years old ranging from 23 to 53 years old;in osteotomy group, there were 26 males and 18 females with an average age of (39.32±6.44) ranging from 22 to 56 years old. The operative time of bone extraction, blood loss in donor area, healing time of donor site and postoperative donor site complications were observed and compared between the two groups. VAS score was used to evaluate the pain of donor site in different periods after operation.</p><p><b>RESULTS</b>All patients were followed up for 12 to 24 months, with an average of 16.9 months in drilling group and 17.1 months in osteotomy groups. The bone healing structure was displayed in the recipient area in two groups, the effect of autogenous iliac bone grafting was good. There was no significant difference in operative time between two groups (>0.05). There was significant difference between two groups in the amount of donor site bleeding and the time of donor site wound healing(<0.05). Postoperative complications(iliac depression and numbness) were significantly different between two groups (<0.05). There was no significant difference in VAS score between two groups at 2 weeks after operation(>0.05). VAS scores of drilling group at 6 months and 1 year after operation were lower than those of osteotomy group (1.85±0.61 vs 2.97±0.67, 0.000; 1.15±0.56 vs 2.41±0.61, 0.000).</p><p><b>CONCLUSIONS</b>When bone graft is no need to have large pieces of special shape or more cortical bone iliac, it is simple to operate and less complications postoperative by drilling type columnar autogenous iliac bone graft. What's more, it has the obvious advantages of promote healing, improve patient quality of life compared with traditional osteotomy.</p>

Pim-1 kinase inhibitor SMI-4a inhibits proliferation and induces apoptosis in U937 cells / 中国病理生理杂志

AIM:To study the growth-inhibiting and proapoptotic effects of Pim-1 kinase inhibitor SMI-4a on human acute myeloid leukemia cell line U937.METHODS:The effect of SMI-4a on U937 cell viability was measured by CCK-8 assay.The apoptotic rate was assessed by flow cytometry with Annexin V-PI staining and by fluorescence microscopy with Hoechst 33342 staining.Methylcellulose was used to assess colony formation ability of the cells.The expression of β-catenin in the cell cytosol and nucleus was detected by Western blot,and the expression of apoptosis-related proteins in the U937 cells was also examined.Intracellular distribution of β-catenin was detected by the method of immunofluorescence.RESULTS:SMI-4a inhibited the viability of U937 cells.Annexin V-PI staining showed that SMI-4a induced apoptosis in dose-and time-dependent manners.Hoechst 33342 staining also verified the apoptosis.SMI-4a significantly inhibited the colony formation capacity of the U937 cells.The results of Western blot demonstrated that SMI-4a upregulated the expression of PARP and Bax,downregulated the expression of Bcl-2 and change the distribution of β-catenin in intracellular compartment.Immunofluorescence observation found that SMI-4a decreased the expression level of β-catenin in the U937 cells.CONCLUSION:SMI-4a induces U937 cell apoptosis through regulating the expression of apoptosis-related genes.

Paraquat promotes epithelial-mesenchymal transition of alveolar epithelial cells through regulating Wnt/β-catenin signaling pathway / 局解手术学杂志

Objective To investigate whether paraquat (PQ) induced rat alveolar type Ⅱ cells (RLE-6TN) epithelial-mesenchymal transition(EMT) and explore the underlying molecular mechanism.Methods RLE-6TN cells were treated by 20 μmol/L PQ for 24 hours,the morphology was observed by invert microscope.RT-PCR and Western blot were performed to detect the expression level of β-catenin,EMT related markers E-cadherin and vimentin.Then we performed the Transwell invasion assays to detect the ability of cell invasion.Results The results demonstrated that PQ was able to induce the transition of RLE-6TN cells from epithelial morphology to fibroblast-like morphology,associated with the acquisition of migratory properties.Furthermore,knockdown of β-catenin by using specific siRNA could reverse PQ triggered EMT process and attenuated cell migration ability.Conclusion PQ promotes RLE-6TN epithelial-mesenchymal transition by upregulating the expression of Wnt/β-catenin.

Pim-1 kinase inhibitor SMI-4a inhibits proliferation and induces apoptosis in U937 cells / 中国病理生理杂志

AIM:To study the growth-inhibiting and proapoptotic effects of Pim-1 kinase inhibitor SMI-4a on human acute myeloid leukemia cell line U937.METHODS:The effect of SMI-4a on U937 cell viability was measured by CCK-8 assay.The apoptotic rate was assessed by flow cytometry with Annexin V-PI staining and by fluorescence microscopy with Hoechst 33342 staining.Methylcellulose was used to assess colony formation ability of the cells.The expression of β-catenin in the cell cytosol and nucleus was detected by Western blot,and the expression of apoptosis-related proteins in the U937 cells was also examined.Intracellular distribution of β-catenin was detected by the method of immunofluorescence.RESULTS:SMI-4a inhibited the viability of U937 cells.Annexin V-PI staining showed that SMI-4a induced apoptosis in dose-and time-dependent manners.Hoechst 33342 staining also verified the apoptosis.SMI-4a significantly inhibited the colony formation capacity of the U937 cells.The results of Western blot demonstrated that SMI-4a upregulated the expression of PARP and Bax,downregulated the expression of Bcl-2 and change the distribution of β-catenin in intracellular compartment.Immunofluorescence observation found that SMI-4a decreased the expression level of β-catenin in the U937 cells.CONCLUSION:SMI-4a induces U937 cell apoptosis through regulating the expression of apoptosis-related genes.

Clinical characteristic analysis of 5 patients with confirmed ebola virus disease / 第二军医大学学报

Objective To analyze the clinical characteristics and prognoses of patients with confirmed Ebola virus disease (EVD) and to summarize the clinical treatment experience. Methods The epidemiologic history, symptoms, signs, treatment, and prognoses of 5 confirmed EVD cases were summarized. And the relationship between clinical features and clinical outcomes was analyzed. Results The 5 patients, 2 men and 3 women, ranged in age from 32-58 years old, with a median age of 46. Three were severe cases when they were admitted while the others were relatively mild. All the patients admitted the exposure history within 21 days before onset of the symptoms, with two patients (one medical staff) from the same transmission chain. The main symptoms included fever, weakness or fatigue, lack of appetite, diarrhea and red eyes, with one patient having gastrointestinal bleeding. The first test of PCR Ebola virus (EBOV) RNA was positive in all the peases. They were promptly isolated and treated with antipyretic and fluid replacement therapy. Meanwhile, prevention of Cplications and treatment of basic diseases were also conducted. Three patients survived at last and the other two died. Conclusion Prompt isolation of the infection source and tracing the contacts can effectively prevent the transmission of EVD-VpCR assay is an effective, rapid and simple method to diagnose EVD. Early supporting treatment-centered multimodality therapy can improve the treatment outcome of EVD.

Effect of silencing HlF-1αby siRNA on the expression of erythropoietin in malignant melanoma of the choroid cell under hypoxia / 国际眼科杂志(Guoji Yanke Zazhi)

AlM: To silent hypoxia inducible factor-1α ( HlF-1α) gene in malignant melanoma of the choroid cell by small interference RNA ( siRNA ) and investigate its effect on the expression of matrix metalloproteinase-2 ( MMP-2 ) in the choroid cell line human uveal melanoma cell (OCM-1) in hypoxia environment.METHODS:OCM-1 cells cultured on culture flask were divided into normal group and hypoxia group. Hypoxia group were divided into five groups: simple hypoxic group, and interference group, and negative control group, and positive control group, and liposome group. Normal group cells were cultured on DMEM culture flask with 10% FBS, 100U/mL penicillin, 100μg/mL streptomycin as well as high concentration of glucose. The cells were maintained at 37℃ in a humidified 5% CO2 incubator. Cells in good condition were selected for experiment. For hypoxia group, chemical hypoxia inducer CoCl2 was added into nutrient medium at the concentration of 100μmol/L to simulate hypoxia microenvironment. We designed and synthesised siRNA ( siRNA + negative control+positive control ) , the target sequences of the HlF-1α to transfect hypoxic malignant melanoma of the choroid cell. SiRNA including HlF-1α siRNA, β-actin siRNA and negative control group synthesized in vitro transfected hypoxic OCM - 1 cell through Lipofectamine2000. The expression of HlF-1α, MMP-2 gene and the protein were detected by RT-PCR and Western blot. RESULTS: Compared with the normal group, the expression of HlF-1α mRNA was not obviously changed (P>0. 05), but the expression of HlF-1α protein and MMP- 2 mRNA protein was significantly higher ( P0. 05).CONCLUSlON: Hypoxia status may upregulate the HlF-1α in OCM-1 cells by increasing the expression of protein. Hypoxia can also inactivate MMP-2, resulting in upregulation of MMP-2 RNA and the expression of its protein. The expression of HlF-1α and MMP-2 mRNA can be down-upregulated by transfecting OCM-1 with HlF-1α siRNA.

Efficacy observation of post-stroke dysphagia treated with acupuncture at Lianquan (CV 23) / 中国针灸

<p><b>OBJECTIVE</b>To verify the clinical efficacy on post-stroke dysphagia treated with acupuncture at Lianquan (CV 23).</p><p><b>METHODS</b>One hundred and eighty patients were randomized into an acupuncture A group, an acupuncture B group and a rehabilitation group, 60 cases in each one. On the basis of the conventional medication, in the acupuncture A group, acupuncture was applied at Lianquan (CV 23); in the acupuncture B group, acupuncture was applied at Hegu (LI 4) and Neiguan (PC 6) and in the rehabilitation group, the swallowing rehabilitation training was adopted. The treatment was given once a day, 5 times a week, and the 4 weeks of treatment was required in all of the groups. The national institute of health stroke scale (NIHSS) and TV X-ray fluoroscope swallowing scale (VFSS) were used to evaluate neurologic deficit and swallowing function before and after treatment in the patients of each group. The morbidity of pneumonia and clinical efficacy were compared among the groups.</p><p><b>RESULTS</b>The scores of NIHSS and VFSS were improved apparently after treatment in the patients of the three groups (all P < 0.05) and the results in the acupuncture A group were superior to those in the other two groups (all P < 0.05). The morbidity of pneumonia in the acupuncture A group was lower than that in the acupuncture B group and the rehabilitation group [3.3% (2/60) vs 6.7% (4/60), 8.3% (5/60), both P < 0.05]. The effective rate in the acupuncture A group was better than that in either of the other two groups [95.0% (57/60) vs 81.7% (49/ 60), 75.0% (45/60), both P < 0.05].</p><p><b>CONCLUSION</b>On the basis of the conventional medication, acupuncture at Lianquan (CV 23) effectively improves the swallowing function, relieves neurological deficit and reduces the morbidity of pneumonia in the patients of post-stroke dysphagia.</p>

Regulation effect of adipose-derived mesenchymal stem cells on pancreatic cancer cells / 中华外科杂志

<p><b>OBJECTIVE</b>To analyze the effects of adipose tissue-derived stem cells (ADSCs) on the proliferation and invasion of pancreatic cancer (PaCa) cells and the the possible mechanism involved.</p><p><b>METHODS</b>ADSCs were isolated and co-cultured with PaCa cells. CCK-8 assay was used to detect the proliferation of PaCa cells. An ELISA was used to determine the concentration of stromal cell-derived factor-1 (SDF-1) in the supernatants. The proliferation of PaCa cells by SDF-1 was measured. AMD3100 regulated the co-culture of ADSCs and PaCa. The tumor growth of PaCa cells was assessed after treatment by ADSCs in vivo.</p><p><b>RESULTS</b>ADSCs can promote the proliferation and invasion of PaCa cells (proliferation: SW1990: 1.535 ± 0.153; PANC-1: 1.370 ± 0.100; the value of control was 1; invasion: SW1990: 47.0 ± 2.6 vs. 28.3 ± 1.3; PANC-1: 40.3 ± 1.8 vs. 24.3 ± 1.3; t = 4.332-9.558, P < 0.05). The expression of SDF-1 was high in ADSCs, but not in PaCa cells (69 ± 5 vs. 0 and 0, F = 389.134, P < 0.01). The promotion of SDF-1 on PaCa cells depends on the concentration. AMD3100 significantly downregulates these growth-promoting effects of ADSCs on PaCa cells. ADSCs significantly promoted the growth of SW1990 in nude mice at the 5(th) week (volume: (1295 ± 102) mm(3) vs. (967 ± 81) mm(3), t = 5.614, P < 0.05) , but not in PANC-1 cell.</p><p><b>CONCLUSION</b>ADSCs can promote the proliferation and invasion of PaCa cells, which may involve the SDF-1/CXCR4 axis.</p>

Quality assessment of clinical guidelines in China: 1993 - 2010 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Clinical practice guidelines (CPGs) play an important role in healthcare in China as well as in the world. However, the current status and trends of Chinese CPGs are unknown. The aim of this study was to systematically review the present situation and the quality of Chinese CPGs published in the peer-reviewed medical literature.</p><p><b>METHODS</b>To identify Chinese CPGs, a systematic search of relevant literature databases (CBM, WANFANG, VIP, and CNKI) was performed for the period January 1978 to December 2010. We used the AGREE II instrument to assess the quality of the included guidelines.</p><p><b>RESULTS</b>We evaluated 269 guidelines published in 115 medical journals from 1993 to 2010 and produced by 256 different developers. Only four guidelines (1%) described the systematic methods for searching and selecting the evidence, 14 (5%) guidelines indicated an explicit link between the supporting evidence and the recommendations, only one guideline used the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system. Thirty-one guidelines (12%) mentioned updates and the average frequency of update was 5.5 years; none described a procedure for updating the guideline. From the assessment with the Appraisal of Guidelines for Research and Ecaluation II (AGREE II), the mean scores were low for the domains "scope and purpose" (19%) and "clarity of presentation" (26%) and very low for the other domains ("rigour of development" 7%, "stakeholder involvement" 8%, "applicability" 6% and "editorial independence" 2%).</p><p><b>CONCLUSIONS</b>Compared with other studies on the quality of guidelines assessed with the AGREE instrument in other countries, Chinese CPGs received lower scores, which indicates a relatively poor quality of the guidelines. However, there was some increase over time.</p>

Leucine zippers improves protein splicing-mediated coagulation factor VIII gene delivery by dual-vector system / 药学学报

In our recent study by exploring an intein-based dual-vector to deliver a B-domain-deleted FVIII (BDD-FVIII) gene, it showed that covalently ligated intact BDD-FVIII molecules with a specific coagulant activity could be produced from expressed heavy and light chains by protein trans-splicing. Here, we assessed the hypothesis that the efficiency of trans-splicing may be increased by adding to the intein sequences a pair of leucine zippers that are known to bring about specific and strong protein binding. The intein-fused heavy and light chain genes were co-transferred into cultured COS-7 cells using a dual-vector system. After transient expression, the intracellular BDD-FVIII splicing was observed and the spliced BDD-FVIII and bioactivity secreted to culture media were quantitatively analyzed. An enhanced splicing of BDD-FVIII with decreased protein precursors from gene co-transfected cells was observed by Western blotting. The amount of spliced BDD-FVIII and bioactivity secreted to the culture media were 106 +/- 12 ng x mL(-1) and 0.89 +/- 0.11 U x mL(-1) analyzed by ELISA and Coatest method respectively, which was greater than leucine zipper free intein-fused heavy and light chain genes co-transfected cells (72 +/- 10 ng x mL(-1) and 0.62 +/- 0.07 U x mL(-1)). The activity of cellular mechanism-independent protein splicing was also improved, as showed by the increasing of spliced BDD-FVIII and bioactivity in culture media from combined cells separately transfected with heavy and light chain genes which was 36 +/- 11 ng x mL(-1) and 0.28 +/- 0.09 U x mL(-1). It demonstrated that the leucine zippers could be used to increase the efficiency of protein trans-splicing to improve the efficacy of a dual-vector mediated BDD-FVIII gene delivery by strengthening the interaction between the two intein-pieces fused to heavy and light chains. It provided evidence for further study in animal model using a dual-adeno-associated virus vector to deliver FVIII gene in vivo.

Trans-splicing of Cys mutated coagulation factor VIII / 药学学报

To investigate the improving effect of inter-chain disulfide formation on protein trans-splicing, we introduce a Cys point mutation at Tyr(664) in heavy chain and at Thr(1826) in light chain of B-domain-deleted FVIII (BDD-FVIII). By co-transfection of COS-7 cell with the two Cys mutated chain genes, the intracellular protein splicing, inter-chain disulfide formation, secreted BDD-FVIII and bioactivity in culture supernatant were observed. The data showed that a strengthened spliced BDD-FVIII with an inter-chain disulfide detected by Western blotting and an elevated secretion of spliced BDD-FVIII (128 +/- 24 ng mL(-1)) compared to control (89 +/- 15 ng mL(-1)), assayed by a sandwich ELISA. A Coatest was performed to assay the secretion of bioactivity in culture supernatant and shown a much higher value (0.94 +/- 0.08 u mL(-1)) compared to that of control (0.62 +/- 0.15 u mL(-1)). It suggests that inter-chain disulfide formation could improve protein trans-splicing based dual-vector delivery of BDD-FVIII gene providing experimental evidence for ongoing in vivo study.

Anti-tumor effect of suicide gene therapy using chimeric promoter plus radiotherapy on cancer cell lines / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the synergistic anti-tumor effect of radiotherapy and horseradish peroxidase/prodrug indole-3-acetic acid (HRP/IAA) gene therapy system using chimeric hTERT promoter responsive to ionizing radiation.</p><p><b>METHODS</b>The synthetic hTERT promoters containing four tandem-repeat copies of radio-inducible CArG elements, and the chimeric promoter containing cytomegalovirus (CMV) early promoter were both constructed. The activities of the chimeric promoters in cancer cell lines (HeLa, A549, and MHCC97) and normal cell line (MRC-5) were detected using luciferase reporter gene expression analysis after a (60)Co γ-irradiation treatment at a series of doses(a single dose of 0 to 10 Gy). The anti-tumor effect of combining irradiation with HRP/IAA gene-directed enzyme prodrug therapy system controlled by the chimeric promoter was tested by colony formation assay, cell counting and apoptosis analysis.</p><p><b>RESULTS</b>The chimeric promoters were ineffective in normal human cells, even after irradiation, but the expression of luciferase gene in tumor cells was significantly higher. The activity of the chimeric promoter in MRC-5 cells was 22.3%, 12.9% and 13.6% of that in HeLa, A549 and MHCC97 cells, respectively. After irradiation, the ratios were 11.7%, 8.7% and 8.8%, respectively. Furthermore, the chimeric promoters could successfully induce the expression of luciferase gene following different doses of radiation, with maximal inducible activity seen after 6 Gy irradiation. The chimeric promoter containing four tandem-repeat copies of radio-inducible CArG elements and CMV early promoter showed the highest activity with 6 Gy irradiation. The relative luciferase activities in HeLa, A549 and MHCC97 cells were 1.7 ± 0.2, 2.3 ± 0.2 and 2.3 ± 0.1, respectively. The chimeric promoter mediated suicide gene therapy system could increase radio-sensitivity in different cancer cells. Compared with the control system, it plus irradiation showed stronger cell proliferation inhibition, 67.3% vs. 26.1% in HeLa, 69.0% vs. 28.3% in A549, 64.6% vs. 20.8% in MHCC97 cells, and also higher apoptosis-inducing effect, 39.6% vs. 14.2% in HeLa, 33.0% vs. 12.4% in A549, and 33.2% vs. 14.2% in MHCC97 cells.</p><p><b>CONCLUSIONS</b>Chimeric promoter containing hTERT promoter, CArG elements and CMV promoter preserve the tumor-specificity in telomerase-positive tumor cells, and irradiation-responsive to low dose of radiation. The suicide gene therapy using this promoter plus radiotherapy show a strong anti-tumor effect in vitro. It is expected to have a good potential for future application in gene radiotherapy.</p>

Enhancing effect of deoxynivalenol-mediated GRP78 down-regulation on heavy chain secretion and bioactivity of two-chain FVIII gene co-transfected cells / 药学学报

Although two chain transfering separately could be used to overcome the volume limitation of adeno-associated virus vectors (AAV) in coagulation factor VIII (FVIII) gene delivery, it leads to chain imbalance for inefficient heavy chain secretion. In this study we aimed to improve the efficacy of two chain strategy in FVIII gene delivery through the degradation of glucose-regulated protein 78 (GRP78) known as a protein chaperone in endoplasmic reticulum (ER) by deoxynivalenol (DON) to decrease GRP78-bound FVIII heavy chain. By treating the two-chain gene transduced 293 cells with DON, the heavy chain (HC) secretion and FVIII bioactivity were observed. Data showed that 293 cells after three hours post-treatment with DON at a concentration of 500 ng mL(-1) resulted in obvious decrease the level of GRP78 but no effect on the cell proliferation. The HC secreted from DON-treated cells transfected with HC gene alone was 59 +/- 11 ng mL(-1), higher than that secreted by control cells (15 +/- 4 ng mL(-1)), and the HC secretion was further increasing to 146 +/- 34 ng mL(-1) in light chain (LC) gene co-transfected cells with an activity measured up to 0.66 +/- 0.15 U mL(-1), also greater than control cells (76 +/- 17 ng mL(-1) and 0.35 +/- 0.09 U mL(-1)). Taken together, these data suggest that DON-mediated GRP78 down-regulation could improve the efficacy of two-chain FVIII gene transfering by facilitating HC secretion, providing an experimental basis for in vivo dual-AAV application in FVIII gene delivery.

Protective effect of ischemia postconditioning on reperfusion injury in patients with ST-segment elevation acute myocardial infarction / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the effect of ischemia postconditioning during the first minutes of reperfusion for the myocardial reperfusion injury in ST-segment elevation acute myocardial infarction (STEMI) patients undergoing emergency percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>STEMI patients undergoing emergency PCI in affiliated hospital of Beihua University between October 2006 and January 2009 were randomly divided into two groups: the control group (n = 34) without any intervention after PTCA, and the postconditioning group (n = 30) with ischemia postconditioning within first minutes of reflow by 3 episodes of 30-second inflation and 30-second deflation with the angioplasty balloon. Reperfusion arrhythmias, CK and CKMB, corrected TIMI frame count (CTFC), wall motion score index (WMSI) and left ventricular ejection fraction (LVEF) by echocardiography were compared between the two groups. MI areas were evaluated with the ECG-54 criteria/32 system and myocardial blush grade (MBG) was measured.</p><p><b>RESULTS</b>The incidence of reperfusion arrhythmias-frequent ventricular premature (26.7% vs. 52.9%) and short array ventricular tachycardia beat (23.3% vs. 58.8%) as well as values of peaks CK [(1162 ± 548) U/L vs. (1732 ± 480) U/L, P < 0.01], CKMB [(165 ± 70) U/L vs. (280 ± 99) U/L, P < 0.01], CTFC (22.23 ± 3.81 vs. 26.97 ± 3.42), WMSI (1.27 ± 0.52 vs. 1.82 ± 0.83), and infarction areas determined by ECG methods (10.60% ± 4.97% vs.14.65% ± 6.88%, all P < 0.05) were all significantly lower in the postconditioning group than in control group while LVEF (0.55 ± 0.08 vs. 0.47 ± 0.10) and MBG (2.27 ± 0.64 vs. 1.47 ± 0.61, all P < 0.05) were significantly higher in the postconditioning group than in control group.</p><p><b>CONCLUSIONS</b>Ischemia postconditioning can significantly reduce myocardial reperfusion injury in patients with STEMI.</p>

Clinical and molecular biologic characteristics of 36 cases of leukemia with 11q23/mll / 中国实验血液学杂志

This study was aimed to analyze the clinical and cytogenetic characteristics of acute leukemia with 11q23/mll rearrangement and explore the reasonable therapeutic principles. Characteristics in general situation, morphology, immunology, molecular biology, cytogenetics, treatment and overall survival of 36 cases of acute leukemias with mll gene rearrangement were studied and analyzed. The results showed that 36 cases with mll gene rearrangement were found positive (7.2%) in 494 patients with acute leukemia. Among the 36 cases of mll rearrangement positive, 32 cases were diagnosed as acute myeloid leukemia (AML) with myeloid antigen expression, of which 5 cases expressed lymphoblastic differentiation antigen; 4 cases were classified as B-lineage acute lymphoblastic leukemia (ALL), of which non-lineage myeloid expression pattern were found in 3 cases. In 29 out of 36 cases (80%) the clonal chromosomal aberration were detected, of which chromosome 11 aberration were observed in 22 cases. All patients received chemotherapy with a total response rate of 47.2%. Of the responded patients, 10 cases relapsed within 6 months, with a recurrence rate of 40%; 9 cases received hematopoietic stem cell transplantation (HSCT), 7 cases of which survived after transplantation. The median survival time of 36 cases was 16 months (range 2 - 46) and their 2-year overall survival rate was 41.4%. The 2-year overall survival rate of 9 patients who received HSCT was 87.5%. It is concluded that acute leukemia patients with mll gene rearrangement show poor response to chemotherapy, high recurrence rate and poor prognosis. Hematopoietic stem cell transplantation may be a reasonable treatment principle to improve these patients' survival situation.