Migration of eleven DSD-FWAs from packaging paper to food / 预防医学

Objective@#To observe the migration of 4,4＇-diaminostilbene-2,2＇-disulfonc acid-based fluorescent whitening agents ( DSD-FWAs ) in food packaging paper, so as to provide evidence for quality and safety supervision for paper packaging materials.@*Methods@#Forty-one paper samples with DSD-FWAs positive were made into 6 cm2 pieces and were soaked in four food simulants ( distilled water, 3% acetic acid, 10% ethanol and 95% ethanol, 10 mL each ). The experiment was carried out at the specified soaking temperature and time. The migration amounts of eleven DSD-FWAs were detected by high performance liquid chromatography-fluorescence detection. @*Results@#C.I.220, C.I.24, C.I.210, C.I.85, C.I.113, C.I.264, C.I.353 and C.I.357 were found in all the four food simulants. At the same time and temperature, the migration amount was highest in 10% ethanol, followed by distilled water, 3% acetic acid and 95% ethanol. C.I.220 was dissolved in all four food simulants, in the range of 20-90 ℃, the migration amount increased with soaking temperature; at 20 ℃, 40 ℃ and 60 ℃, the migration amount increased first and then stabilized over time.@*Conclusion@#The higher the storage temperature and the longer the storage time of paper packaging, the easier the DSD-FWAs in packaging paper migrate to food.

Exercise behavior and its relevant factors in 3 099 pregnant women during pregnancy / 上海预防医学

Objective To investigate the current situation of the exercise, relevant knowledge and impact factors in pregnant women during pregnancy, providing references to instruct pregnant women to do suitable exercise. Methods Self-designed questionnaires were used to investigate the exercise behavior and cognition in 3 099 pregnant women who visited the clinic for first-time prenatal examination at Jiading District of Shanghai. Results It was found in investigation that 65.21% pregnant women often did exercise, of whom 29.14% did less than 20 minutes exercise every time, while 60.96% did 20 to 40 minutes, and 9.90% did more than 40 minutes.Slow-walking was the majority choice, which counted for 87.14% in pregnant women.And 34.79% pregnant women did not do exercise, mainly due to "wouldn't like to do exercise" or "don't have time for it".Age, body mass index (BMI), educational level with the pregnant women as well as their husbands impact factors related to their exercise behavior during pregnancy (P<0.01). Conclusion At present, pregnant women exercise mode is unitary during pregnancy.Doctors and nurses should advocate more on health education for them, and family members should be involved in health education program as well.We should improve the education health system and give full play to the advantages of the Internet, train obstetricians, nutritionists, community doctors and other weight management professionals, improve service capabilities, and instruct pregnant women to choose safe and effective exercise during pregnancy.

Exercise behavior and its relevant factors in 3 099 pregnant women during pregnancy / 上海预防医学

Objective To investigate the current situation of the exercise, relevant knowledge and impact factors in pregnant women during pregnancy, providing references to instruct pregnant women to do suitable exercise. Methods Self-designed questionnaires were used to investigate the exercise behavior and cognition in 3 099 pregnant women who visited the clinic for first-time prenatal examination at Jiading District of Shanghai. Results It was found in investigation that 65.21% pregnant women often did exercise, of whom 29.14% did less than 20 minutes exercise every time, while 60.96% did 20 to 40 minutes, and 9.90% did more than 40 minutes.Slow-walking was the majority choice, which counted for 87.14% in pregnant women.And 34.79% pregnant women did not do exercise, mainly due to "wouldn't like to do exercise" or "don't have time for it".Age, body mass index (BMI), educational level with the pregnant women as well as their husbands impact factors related to their exercise behavior during pregnancy (P<0.01). Conclusion At present, pregnant women exercise mode is unitary during pregnancy.Doctors and nurses should advocate more on health education for them, and family members should be involved in health education program as well.We should improve the education health system and give full play to the advantages of the Internet, train obstetricians, nutritionists, community doctors and other weight management professionals, improve service capabilities, and instruct pregnant women to choose safe and effective exercise during pregnancy.

Determination of vanadium, chromium, nickel, selenium and arsenic in calcium based toothpastes by microwave digestion-ICP-MS/MS / 预防医学

Objective@#To establish analytical method for the determination of vanadium ( V ), chromium ( Cr ), nickel ( Ni ), selenium ( Se ) and arsenic ( As ) in calcium based toothpastes by microwave digestion-inductively coupled plasma tandem mass spectrometry ( ICP-MS/MS ).@*Methods@#The 21 calcium based toothpaste samples from supermarkets and shops in the urban areas of Hangzhou were digested by 6 mL HNO3 and 1 mL H2O2 in microwave digestion system. Then He-SQ mode and O2-MS/MS mode of ICP-MS/MS were respectively used for the determination of Ni and V, Cr, Se, As. Indium ( In ) was used as internal standard for calibration. @*Results@# Good linear relationships were obtained for these five elements from 1.0 to 32.0 μg/L, with the correlation coefficients ranged from 0.999 3 to 1.000 0. The detection limits of the method ranged from 0.000 25 to 0.006 08 mg/kg. The recovery rates of standard spiking were 80.7%-105.7% when set at 0.4, 0.8 and 1.6 mg/kg, the recovery of standard reference material was 102.2%, and the relative standard deviations were 2.6%-4.8%. The concentrations of V, Cr, Ni, Se and As in 21 calcium based toothpaste samples were 0.024-1.935 mg/kg, 0.085-5.759 mg/kg, 0.090-3.673 mg/kg, <0.002 72-0.016 mg/kg and <0.006 08-0.321 mg/kg.@*Conclusion@#Microwave digestion-ICP-MS/MS can effectively reduce the interferences of polyatomic ions and doubly charged ions from the matrix, which is suitable for the determination of V, Cr, Ni, Se and As in calcium based toothpaste.

Retrovectors packaged in CHO cells to generate GLP-1-Fc stable expression CHO cell lines

Background: Chinese hamster ovary (CHO) cells are the most dependable mammalian cells for the production of recombinant proteins. Replication-incompetent retroviral vector (retrovector) is an efficient tool to generate stable cell lines. Multiple copies of integrated genes by retrovector transduction results in improved recombinant protein yield. HEK-293 and their genetic derivatives are principal cells for retrovector production. Retrovectors packaged in HEK-293 cells pose a risk of infectious agent transmission, such as viruses and mycoplasmas, from serum and packaging cells. Results: In this report, retrovectors were packaged in CHO cells cultured in chemically defined (CD) media. The retrovectors were then used to transduce CHO cells. This method can block potential transmission of infectious agents from serum and packaging cells. With this method, we generated glucagon-like protein-1 Fc fusion protein (GLP-1-Fc) stable expression CHO cell lines. Productivity of GLP-1-Fc can reach 3.15 g/L. The GLP-1-Fc protein produced by this method has comparable bioactivity to that of dulaglutide (Trulicity). These stable cell lines retain 95­100% of productivity after 40 days of continuous culture (~48­56 generations). Conclusions: Suspension CHO cells are clean, safe, and reliable cells for retrovector packaging. Retrovectors packaged from this system could be used to generate CHO stable cell lines for recombinant protein expression.

Survey of knowledge and practice of residents about schistosomiasis preven-tion and control in endemic areas of Jingzhou City / 中国血吸虫病防治杂志

Objective To investigate the knowledge and practice of residents about schistosomiasis prevention and control in endemic areas of Jingzhou City,Hubei Province after the disease transmission being controlled,so as to provide the valuable in-formation for formulating an efficient health education and intervention strategy.Methods The residents were selected by using the cluster sampling method and investigated with questionnaires in Jiangling County and Gong'an County,Jingzhou City,Hu-bei Province,and the data were analyzed by using the descriptive analysis method,Chi-square tests and logistic regression.Re-sults In a total,826 available questionnaires were obtained with 100% of valid rate.Among them,97.0% of the interviewee knew schistosomiasis,and 86.3% knew that the infection happened by contacting water containing cercariae.The residents in el-der age(41-84 years)had higher awareness rates than the residents in lower age(6-17 years),about the regions of epidemic area(χ 2=57.860),infection route of schistosomiasis(χ 2=87.045),advanced schistosomiasis symptoms(χ 2=27.268)and On-comelania hupensis being as intermediate host(χ 2=55.856)(all P<0.05).The males had higher awareness rates of epidemic areas than the females(χ2=13.442,P<0.05).For personal behavior,36.6% of the interviewee had experience of contacting lake or pond water many times a day,and 66.5% had the willingness to participate in schistosomiasis health education.Conclu-sion In the investigation areas,Jiangling County and Gong'an County,the awareness rates of the residents about schistosomia-sis prevention and control are higher,and we should strengthen the health education and behavior intervention,especially in the students of middle and primary schools to help them have the self-protection ability efficiently.

Changes of serum anti-schistosome antibody levels in schistosomiasis japoni-ca patients after treatment / 中国血吸虫病防治杂志

Objective To investigate the changes of serum anti-schistosome antibody titers in schistosomiasis japonica pa-tients after treatment,in order to provide the evidence for formulating the schistosomiasis surveillance program in marshland and lake regions.Methods Upon prospective cohort study,the stool examination positive schistosomiasis patients and blood exami-nation positive suspected patients(the titer was more than 1:80,including 1:80)were selected as the research objects in Jian-gling County in 2014,and they received the 2-day praziquantel therapy.Half year,one year and two years after the treatment, their blood samples and fecal samples were collected for IHA anti-schistosome antibody detections and schistosome egg and mira-cidium detections. Results In 2014,the stool examination positives were 251,and the majority of them were over 41 years old,accounting for 93.23%(234/251);581 cases of high antibody titers were detected by the IHA method,and the majority of them were over 41 years old,accounting for 89.16%(518/581).Half year,one year and two years after the treatment,among the stool examination positives,the negative conversion rates of stool positives were 99.60%(250/251),100%(239/239)and 100%(234/234)respectively and the negative conversion rates of antibody positives were 21.91%(55/251),64.11%(156/239)and 76.89%(193/234)respectively.In the high antibody titer positives,the negative conversion rates were 38.04%(221/581),64.11%(359/560),and 77.86%(429/551)respectively,Half year,one year and two years after the treatment.There were statistically significant differences among the antibody negative conversion rates by χ2test(χ2=77.538,183.412,25.469 respectively,all P<0.001).The geometric mean values of antibody titers of different durations between 2 groups were analyzed by 2-independent-samples T test,and the geometric mean values of antibody titers between the 2 groups were different before the treatment(t=23.576,P<0.01),but the geometric mean values of antibody titers between the 2 groups were not different 6 months,1 year and 2 years after the treatment(t=-0.046,1.165, -0.132,P=0.964, 0.245,0.895 respectively). Conclu-sions The levels of serum anti-schistosome antibody degrade slowly in schistosomiasis japonica patients after the treatment, and the results of IHA tests cannot distinguish the current schistosome infection from previous schistosome infection.Therefore, it is necessary to develop the specific diagnostic technology for schistosome infection in order to meet the need of monitoring.

Analysis on centralized procurement and distribution mode of medical consumables under new situation / 医疗卫生装备

Objective To explore the function of medical consumables integrated bidding procurement and the countermeasures of medical institutions under the background of health care reform. Methods The present situation of integrated bidding procurement was analyzed,and its significance and program were described.It's suggested that the whole-course supervision and feedback evaluation be executed on procurement cycle,transaction price.Some countermeasures were proposed to optimize the distribution mode. Results The implementation of centralized bidding procurement of medical consumables helped to integrate market resources, reduce procurement and distribution costs, strengthen supply chain management and intensify market competition. Conclusion The promotion of centralized bidding procurement and distribution mode of medical consumables nationally is the inevitable trend of medical reform, which is beneficial to the benign development of the industry and ultimately benefits the patients.

The anomalies of choroidal and retinal blood flow in cone-rod dystrophy / 眼科新进展

Objective To study the alteration of blood flow indexes of ophthalmic artery (OA),central retinal artery (CRA) and posterior ciliary artery (PCA) of the patients with cone-rod dystrophy (CRD).Methods The color doppler flow imaging was used in 50 CRD patients (100 eyes,CRD group) and 90 controls (180 eyes,control group) to measure the peak systolic velocity (PSV),end diastolic velocity (EDV),timeaveraged maximum velocity (TAMV),pulsatility index (PI) and resistance index (RI)in OA,CRA and PCA,respectively,and the data was obtained and analyzed statistically by one-sample t test to compare the alteration of these parameters between the CRD group and control group.Results The TAMV of OA was (14.29 ±3.88)cm · s-1 in CRD group and (12.44 ± 3.64) cm · s-1 in the control group,and the difference was statistically significant (P ＜0.05);meanwhile,the PI was 1.75 ±0.42 in CRD group and 2.02 ±0.71 in the control group,and the difference was statistically significant (P ＜0.05).Moreover,the PSV,EDV,TAMV and PI,RI of CRA was (4.60 ± 1.29) cm · s-1,(1.61 ±0.41)cm · s-1,(2.59 ±0.67)cm · s-1 and 1.11 ±0.31,0.63 ±0.10 in CRD group,respectively,and (10.82 ± 2.97) cm · s-1,(3.28 ± 1.11) cm · s-1,(5.50 ± 2.06)cm · s-1 and 1.48 ±0.49,0.71 ±0.08,in the control group,accordingly,and the differences were statistically significant (all P ＜ 0.05).Furthermore,the PSV,EDV,TAMV and PI,RI of PCA was (7.36 ± 2.18) cm · s-1,(2.28 ± 0.82) cm · s-1,(3.99 ± 1.22)cm · s-1 and 1.28 ± 0.37,0.68 ± 0.09 in CRD group,respectively,and (11.61 ± 3.41)cm · s-1,(3.34±1.25)cm · s-1,(5.83 ±1.91)cm · s-1 and 1.49 ±0.43,0.70 ±0.09 in the control group,accordingly,and the differences were statistically significant (all P ＜0.05).Conclusion Hemodynamic unusual alterations of OA,CRA and PCA in CRD patients suggest there is the correlation between the ocular blood supply and the pathological process of CRD,which remains to be observed further.

Study on the related factors of lymph node detection number in rectal cancer patients underwent laparoscopic surgery / 中国内镜杂志

Objective To investigate the related factors of lymph node detection number in rectal cancer patients underwent laparoscopic surgery. Methods 98 patients with rectal cancer who underwent laparoscopic surgery were selected from January 2014 to January 2010. All the patients general information [gender, age, body mass index (BMI)], preoperative imaging findings and pathological data (tumor size, gross type, TNM stage, distant metastasis, histological differentiation and depth of invasion, et al), surgery related data (experience of surgeon, operation time) and preoperative radiotherapy and chemotherapy were collected. Results The age, BMI, tumor size, length of specimen, invasive depth, surgeon and preoperative radiotherapy and chemotherapy was correlated with the number of lymph nodes in patients with laparoscopic surgery (P < 0.05), but gender, TNM staging, general type, histological differentiation, operation time were not associated with the number of lymph nodes detected in minimally invasive surgery for rectal cancer (P > 0.05). Multiple linear regression analysis showed that BMI, tumor size, length of specimen, invasive depth, surgeon and preoperative radiotherapy and chemotherapy were the independent influencing factors of lymph node detection in patients with minimally invasive rectal cancer (P < 0.05). Conclusion The factors of patients, tumor status, surgical factors and preoperative chemoradiotherapy are related to the number of lymph nodes in patients with rectal cancer.

Effect of consumption ratio monitoring on management of single disease / 医疗卫生装备

Objective To explore the effect of the consumption ratio monitoring on the management of single disease so as to enhance medical consumables management.Methods The characteristics and functions of single disease management were introduced,and the present situation of the consumption ratio was analyzed by discussing medical consumables management.The consumption ratio was introduced into single disease management to solve the problems in medical consumables management such as high consumption ratio,amplification,extensive management and etc.Results The introduction of the consumption ratio decreased medical cost,use risks,overstock and inventory loss.Conclusion The consumption ratio involved in single disease management enhances medical consumables management greatly.

Prostaglandin E2 Receptor 4 Agonist Promotes Human CD34Cell Proliferation in vitro by Activating Wnt/β-Catenin Signaling Pathway / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the potential signaling pathway that regulates the proliferation of human CD34cells stimulated by prostaglandin E2 receptor 4 agonist (EP4A) in vitro.</p><p><b>METHODS</b>Twenty samples of peripheral blood containing stem cells were collected from the G-CSF mobilized healthy donors in our department of hematology. Human CD34cells were isolated by magnetic activated cell sorting (MACS) microbeads kit. The Cell Counting Kit-8 (CCK8) assay was used to determine the optimal concentration and time of EP4A to promote human CD34cell proliferation in vitro. Under the optimal condition, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect mRNA level of β-catenin, and Western blot was used to assay protein expression of β-catenin and P-GSK-3β in human CD34cells treated with EP4A.</p><p><b>RESULTS</b>Culturing with 10 µmol/L EP4A for 72 h, it was found that EP4A promoted human CD34cell proliferation significantly, and the proliferation rate of human CD34cells was 1.36 times higher than that of the control(P=0.002). Under the optimal condition, it was also found that EP4A enhanced the β-catenin expression at both mRNA and protein levels, and up-regulated phosphorylation of GSK-3β in human CD34cells, but these effects could be inhibited by the EP4A antagonist EP4AA.</p><p><b>CONCLUSION</b>EP4A can enhance human CD34cell proliferation in vitro by activating Wnt/β-catenin signaling pathway.</p>

Effects of dendritic cell-activated and cytokine-induced killer cell therapy on 22 children with acute myeloid leukemia after chemotherapy / 华中科技大学学报（医学）（英德文版）

The efficiency of dendritic cell-activated and cytokine-induced killer cell (DC-CIK) therapy on children with acute myeloid leukemia (AML) after chemotherapy was investigated. Mononuclear cells were collected from children achieving complete remission after chemotherapy, cultured in vitro and transfused back into the same patient. Interleukin-2 (IL-2) was injected subcutaneously every other day 10 times at the dose of 1 × 10(6) units. Peripheral blood lymphocyte subsets and minimal residual disease (MRD) were detected by flow cytometry. Function of bone marrow was monitored by methods of morphology, immunology, cytogenetics and molecular biology. The side effects were also observed during the treatment. The average follow-up period for all the 22 patients was 71 months and relapse occurred in two AML patients (9.1%). The percentage of CD3(+)/CD8(+) cells in peripheral blood of 15 patients at the 3rd month after DC-CIK treatment (36.73% ± 12.51%) was dramatically higher than that before treatment (29.20% ± 8.34%, P < 0.05). The MRD rate was >0.1% in 5 patients before the treatment, and became lower than 0.1% 3 months after the treatment. During the transfusion of DC-CIK, side effects including fever, chills and hives appeared in 7 out of 22 (31.82%) cases but disappeared quickly after symptomatic treatments. There were no changes in electrocardiography and liver-renal functions after the treatment. MRD in children with AML can be eliminated by DC-CIK therapy which is safe and has fewer side effects.

Cloning, characterization and expression of Peking duck fatty acid synthase during adipocyte differentiation

Background Fatty acid synthase (FAS) is a key enzyme of de novo lipogenesis (DNL), which has been cloned from several species: Gallus gallus, Mus musculus, Homo sapiens, but not from Anas platyrhynchos. The current study was conducted to obtain the full-length coding sequence of Peking duck FAS and investigate its expression during adipocyte differentiation. Results We have isolated a 7654 bp fragment from Peking duck adipocytes that corresponds to the FAS gene. The cloned fragment contains an open reading frame of 7545 bp, encodes a 2515 amino acid protein, and displays high nucleotide and amino acid homology to avian FAS orthologs. Twelve hour treatment of oleic acid significantly up-regulated the expression of FAS in duck preadipocytes (P < 0.05). However, 1000 µM treatment of oleic acid exhibited lipotoxic effect on cell viability (P < 0.05). In addition, during the first 24 h of duck adipocyte differentiation FAS was induced; however, after 24 h its expression level declined (P < 0.05). Conclusion We have successfully cloned and characterized Peking duck FAS. FAS was induced during adipocyte differentiation and by oleic acid treatment. These findings suggest that Peking duck FAS plays a similar role to mammalian FAS during adipocyte differentiation.

Clinical significance of abnormal protein bands in multiple myeloma treated with bortezomib-based induction regimen and autologous stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the clinical significance of abnormal protein bands (APB) in multiple myeloma (MM) patients treated with bortezomib-based induction regimen and autologous stem cell transplantation (ASCT).</p><p><b>METHODS</b>Sixty-eight MM patients submitted to bortezomib-based induction therapy and ASCT from January 2007 to July 2012 were retrospectively studied. Monoclonal protein was detected by immunofixation electrophoresis (IFE).</p><p><b>RESULTS</b>Of all 68 patients, 33 (48.5%) patients had APB. At the first emergence of an APB, two patients with light chain type achieved CR and before transplantation, and thirty-one patients were after transplantation with median time of 104 (ranged 33-404) days. The median duration of APB appearance was 105 (ranged 35-801) days. Patients who developed APB compared with those without APB, had a significantly higher CR plus very good partial response (VGPR) rates (100.0% vs 85.7%%, P=0.017) and CR rates (87.9% vs 62.9%) (P=0.03). There were no significant differences in gender, age, HGB, ALB, β2-microglobulin, M protein type, Durie-Salmon and ISS stages, the case number of first line or second line treatment, induction courses of bortezomib-based regimen, and the mode of ASCT. With a median follow-up of 33.4 (ranged 7.0-71.7) months, patients with APB tended to have a longer overall survival (OS) versus non-APB patients, although no significant difference obtained (P＞0.05). Among APB patients, OS was longer in patients whose appearance of APB occurred ＜6 months after transplantation than those ≥ 6 months, but the significant difference was not obtained yet (P＞0.05).</p><p><b>CONCLUSIONS</b>Patients who developed APB had a significantly better response to bortezomib-based induction regimen followed ASCT. APB emergence has a good prognostic significance.</p>

Molecular cloning and in silico analysis of the duck (Anas platyrhynchos) MEF2A gene cDNA and its expression profile in muscle tissues during fetal development

The role of myogenic enhancer transcription factor 2a (MEF2A) in avian muscle during fetal development is unknown. In this work, we cloned the duck MEF2A cDNA sequence (GenBank accession no. HM460752) and examined its developmental expression profiles in cardiac muscle, non-vascular smooth muscle and skeletal muscle. Duck MEF2A cDNA comprised 1479 bp encoding 492 amino acid residues. In silico analysis showed that MEF2A contained MADS (MCM1, AGAMOUS, DEFICIENS and SRF -serum response factor), MEF2 and mitogen-activated protein kinase (MAPK) transcription domains with high homology to related proteins in other species. Modified sites in these domains were conserved among species and several variants were found. Quantitative PCR showed that MEF2A was expressed in all three muscles at each developmental stage examined, with the expression in smooth muscle being higher than in the other muscles. These results indicate that the conserved domains of duck MEF2A, including the MADS and MEF2 domains, are important for MEF2A transcription factor function. The expression of MEF2A in duck smooth muscle and cardiac muscle suggests that MEF2A plays a role in these two tissues.

Blockade of the Sonic Hedgehog Signalling Pathway Inhibits Choroidal Neovascularization in a Laser-induced Rat Model / 华中科技大学学报（医学）（英德文版）

Sonic hedgehog (Shh) signaling has recently been shown to be involved in the pathological angiogenesis in response to tissue hypoxia and ischemic injury. Hypoxia/ischemia is considered to play an important role in the development of choroidal neovascularization (CNV). This study was aimed to examine the effect of blockade of the Shh signaling pathway on CNV and the underlying mechanism. A total of 64 male Brown-Norway (BN) rats were used in this study. One eye of each rat underwent laser photocoagulation. The other eye served as normal control. After the laser treatment,the 64 rats were divided into four groups (n=16 in each group): Blank control group, in which no intravitreal administration was given; cyclopamine group, recombinant Shh N-terminals protein (rShh)group and phosphate-buffered saline (PBS) group, in which cyclopamine (a Shh inhibitor), rShh (a Shh activator) and PBS were intravitreally injected into the laser-treated eyes respectively every other day for a total of four intravitreal injections immediately after the laser treatment. Fourteen days after the intravitreal administration, the changes of CNV-related variables, including positive CNV lesion percentage, CNV membrane area and CNV membrane thickness, were evaluated by fluorescein angiography, indocyanine green angiography and pathological examinations. The mRNA and protein expression of PTCH1, Glil, HIF-la, VEGF and DLL4 in each group on 14 days of CNV model was detected by real-time quantitative PCR and western blot analysis, and the relationship between the Shh cascade and the HIF-1 α-VEGF-DLL4 cascade in CNV was analyzed. The results showed that the CNV membrane area and the CNV membrane thickness were decreased by 62.5% and 41.9% in the cyclopamine group and increased by 85.7% and 64.3% in the rShh group in comparison to those in the blank control group (P＜0.01 for each). There was no significant difference in the CNV membrane area and thickness between the blank control group and PBS group (P=0.102 and P=0.063, respectively). Real-time quantitative PCR revealed a 5.23-, 4.14-, 2.97-, 2.78- and 2.39-fold up-regulation of the mRNA expression of PTCH1, Glil, HIF-la, VEGF and DLL4 genes in the laser-treated eyes compared with the normal control eyes in the control group. In the cyclopamine group, the mRNA and protein expression of Glil, HIF-lα, VEGF and DLL4 was significantly down-regulated (P＜0.05 for each) while the expression of PTCH1 showed no significant changes at the mRNA (P=0.293) and protein level (P=0.304). The mRNA expression and protein expression (P=0.001 and P=0.021, respectively) of PTCH1, Glil, HIF-lα, VEGF and DLL4 was significantly increased in the rShh group when compared with the control group. The expression level of these genes was related to the severity of the CNV. It was concluded that intravitreal administration of cyclopamine can effectively inhibit the formation of laser-induced experimental CNV by down-regulating the expression of the HIF-la-VEGF-DLL4 cascade in CNV. The Shh signaling pathway as an upstream signaling pathway of HIF-1 a-VEGF-DLL4 cascade is implicated in the development of experimental CNV.

Effects of killer immunoglobulin-like receptor and human leukocyte antigen class I ligand on the prognosis of related donor hematopoietic stem cell transplantation / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the genotype distribution and the effects of killer immunoglobulin-like receptors (KIR) and human leukocyte antigen (HLA) class I ligand on related donor hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>The genotypes of donor/recipient HLA-Cw and donor KIR were determined by polymerase chain reaction-sequence specific primer (PCR-SSP) in 87 cases of related donor HSCT (40 cases were haploidentical HSCT, and the remaining 47 cases were HLA-identical sibling HSCT).</p><p><b>RESULTS</b>All the donors possessed KIR2DL1, 2DL2/L3, 2DL4, 3DL2, and 3DL3, and 96.6% of donors possessed 3DL1. The rate of activating KIRs varied. 97.7% of the recipients expressed C1, while the rates of C2, Bw4, and HLA-A3/A11 were different. In haploidentical HSCT, KIR-HLA-mismatched group included 34 cases and the matched group included 6 cases. HLA-HLA-mismatched group included 31 cases and the matched group included 9 cases. In matched sibling donor HSCT, KIR-HLA-mismatched group included 42 cases and the matched group included 5 cases. KIR-HLA-mismatched group had higher 2-year disease-free survival (DFS) rate compared with KIR-HLA-matched group [ (71.5 +/- 6.5 ) % vs. (50.0 +/- 10.7)%, P < 0.05].</p><p><b>CONCLUSIONS</b>The rate of activating KIR is lower than inhibitory KIRs. Inhibitory KIR2DL1, 3DL1, and 3DL2 may play key roles in the natural killer cell alloreactivity. The DFS rate is higher in KIR-HLA-mismatched group than in KIR-HLA-matched group in related donor HSCT.</p>

Overexpression or knock-down of runt-related transcription factor 1 affects BCR-ABL-induced proliferation and migration in vitro and leukemogenesis in vivo in mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Runt-related transcription factor 1 (Runx1) plays a crucial role in hematogenesis and its dysfunction may contribute to leukemogenesis. However, it is not clear whether or not abnormal expression of Runx1 will induce leukemia and how the change of Runx1 expression level could affect BCR-ABL-induced leukemogenesis. In the present study, we aimed to analyze if abnormal expression of Runx1 in BaF3 cells alone would induce leukemogenesis. And we also wanted to know if abnormal expression of Runx1 in leukemic cells would affect leukemogenesis. Furthermore, we investigated whether overexpression or knock-down of Runx1 in BaF3 cells would induce leukemogenesis.</p><p><b>METHODS</b>Plasmids containing full-length Runx1 cDNA were transduced into BaF3 cells and BaF3-P185wt cells (BCR-ABL transformed BaF3 cells) by electroporation. Plasmids containing a short hairpin RNA of Runx1 were transduced into BaF3 cells and BaF3-P185wt cells by electroporation. Runx1 expression level was quantified by Western blotting and quantitative real-time PCR. The effects of overexpression or knock-down of Runx1 on proliferation, apoptosis and migration of cells were detected in vitro. Then, using MSCV-P185wt-EGFP as a control, we transplanted MSCV-P185wt-Runx1 cells or MSCV-P185wt-shRNA cells into Balb/c mice through tail vein and observed tumorgenesis of the different phenotypes.</p><p><b>RESULTS</b>In vitro analysis revealed that overexpression of Runx1 in P185wt cells could inhibit cell proliferation and slow down cell migration; while knock-down of Runx1 could promote cell proliferation and speed up cell migration. In vivo analysis indicated that mice transplanted with MSCV-P185wt-Runx1 survived longer than controls. In contrast, mice transplanted with MSCV-P185wt-shRNA survived shorter than the control group. Gross pathological analysis revealed that the MSCV-P185wt-Runx1 group had less severe splenomegaly and hepatomegaly compared to the control group, and the MSCV-P185wt-shRNA group had more severe splenomegaly and hepatomegaly. No splenomegaly or hepatomegaly was detected in mice transplanted with MSCV-BaF3-Runx1 cells or MSCV-BaF3-shRNA cells. Both the mice of MSCV-BaF3-Runx1 group and MSCV-BaF3-shRNA group were healthy with no sign of leukemia for up to three months.</p><p><b>CONCLUSIONS</b>Overexpression or knock-down of Runx1 gene in BaF3 cells alone could not induce leukemogenesis. However, in BaF3-P185wt cells, alteration of Runx1 expression could affect BCR-ABL-induced proliferation and migration in vitro and leukemogenesis in vivo.</p>

Osteoclast differentiation regulated by receptor activator of nuclear factor kappaB probably through a novel signaling pathway / 中国实验血液学杂志

This study was aimed to investigate the signaling pathways regulating osteoclast (OC) differentiation by receptor activator of nuclear factor kappa (RANK) under physiological condition so as to provide some theoretical basis for clarifying mechanism of bone destruction in multiple myeloma. A mutant TNFR(1)/RANK(2) (named RANK-Mu) chimera consisting of tumor necrosis factor receptor 1 (TNFR(1)) and RANK intramembrane domain was constructed by using deletion mutation for deleting IVVY amino acids in RANK intramembrane domain in accordance with (535-)IVVY(-538) as specific domain regulating OC differentiation by RANK. The RANK-Mu and TNFR(1)/RANK chimera without mutation (RANK-WT) were packaged by using plat E cell line to produce the retrovirus, which were transfected into bone marrow macrophages (BMMs) of TNFR(1)/TNFR(2) double knockout mice. After stimulation of these transfected BMMs with TNF-alpha, the differentiation of BMMs into OCs were observed, meanwhile the phosphorylation of NF-kappab, JNK, p38 and ERK was detected by Western blot after stimulation of these BMMs with TNF-alpha. The results showed that BMMs transfected with RANK-WT could be differentiated into OCs and phosphorylation of NF-kappaB, JNK, P38 and ERK were activated at 5 - 10 minutes after being stimulated by TNFalpha. BMMs transfected with RANK-Mu could not be differentiated into OCs, but phosphorylation of NF-kappaB, JNK, P38 and ERK were activated also. It is concluded that RANK regulates osteoclast differentiation probably not through 4 typical signaling pathways, named as NF-kappaB, JNK, P38 and ERK, in this process other new signaling pathways maybe participate.