Study on the Relationship between Integrin 2A and Drug Resistance in Chronic Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To explore the expression pattern and clinical significance of Integral membrane protein 2A（ITM2A） in drug resistant patients with chronic myeloid leukemia (CML).@*METHODS@#The expression of ITM2A in CML was evaluated by qRT-PCR, Western blot and immunocytochemistry. In order to understand the possible biological effects of ITM2A, apoptosis, cell cycle and myeloid differentiation antigen expression of CML cells were detected by flow cytometry after over-expression of ITM2A. The nuderlying molecular mechanism of its biological effect was explored.@*RESULTS@#The expression of ITM2A in bone marrow of CML resistant patients was significantly lower than that of sensitive patients and healthy donors（P<0.05）. The CML resistant strain cell K562R was successfully constructed in vitro. The expression of ITM2A in the resistant strain was significantly lower than that in the sensitive strain(P<0.05). Overexpression of ITM2A in K562R cells increased the sensitivity of K562R cells to imatinib and blocked the cell cycle in G2 phase(P<0.05), but did not affect myeloid differentiation. Mechanistically, up-regulation of ITM2A reduced phosphorylation in ERK signaling (P<0.05).@*CONCLUSION@#The expression of ITM2A was low in patients with drug resistance of CML, and the low expression of ITM2A may be the key factor of imatinib resistance in CML.

Network Meta-analysis of Chinese patent medicine containing Hirudo in treatment of atherosclerosis / 中国中药杂志

This study aimed to evaluate the efficacy and safety of Chinese patent medicines containing Hirudo in the treatment of atherosclerosis(AS) by network Meta-analysis, and to provide evidence-based reference for clinical treatment of AS. The clinical randomized controlled trial(RCT) on the treatment of atherosclerosis with Chinese patent medicines containing Hirudo were searched in CNKI, Wanfang, VIP, SinoMed, PubMed and EMbase from the establishment of the databases to July 1, 2022. And data extraction and quality assessment of the included RCT was performed according to the Cochrane standards. Stata 17 and ADDIS 1.16.5 were then used for Bayesian model network Meta-analysis. Finally, 67 RCTs with a total sample size of 6 826 cases were included, 3 569 cases in the experimental group and 3 257 cases in the control group, involving three oral Chinese patent medicines. Network Meta-analysis showed that in terms of reducing intima-media thickness(IMT), the top three Chinese patent medicines were Tongxinluo Capsules+sta-tins>Maixuekang Capsules+statins>Maixuekang Capsules. In terms of reducing plaque area, the top one was Maixuekang Capsules+sta-tins, and the other Chinese patent medicines had similar efficacy. For lowering AS Crouse scores, the top three were Maixuekang Capsules>Tongxinluo Capsules+statins>Naoxintong Capsules. For decreasing plaque number, the top three were Naoxintong Capsules+sta-tins>Tongxinluo Capsules+statins>Tongxinluo Capsules. With regard to adverse reactions/events, Naoxintong Capsules+statins had the lo-west incidence. In conclusion, in Chinese patent medicines containing Hirudo for the treatment of AS, Tongxinluo Capsules+statins, Maixuekang Capsules, Maixuekang Capsules+statins, and Naoxintong Capsules+statins were the primary choices to reduce IMT, AS Crouse scores, plaque area, and plaque number, respectively. The efficacy of Chinese patent medicines containing Hirudo with or without statins was more significant than that of statins alone in the four outcome indexes. Additionally, the treatment of AS should be evaluated comprehensively, and attention should be paid to Chinese patent medicines or their combination with western medicine, to optimize the treatment effect and minimize adverse reactions as the benchmark.

Changes of cytokines after cardiopulmonary bypass in children with congenital heart disease / 中国小儿急救医学

Objective:To study the changes of cytokines after cardiopulmonary bypass(CPB)in children with congenital heart disease.Methods:A total of 124 children with congenital heart disease underwent CPB surgery at Shanghai Children′s Medical Center from June 2020 to October 2021 with cytokine detection were enrolled.Twelve kinds of cytokines, white blood cell count(WBC)and neutrophil-to-lymphocyte ratio(NLR), C-reactive protein(CRP)and procalcitonin were detected before and 24 hours after operation.All patients were divided into CPB<120 min group ( n=102)and CPB≥120 min group ( n=22)acoording to CPB time, and were divided into systemic inflammatory response syndrome (SIRS) group, compensatory anti-inflammatory response syndrome (CARS) group and control group according to the changes of cytokines.The changes of cytokines, anti-inflammatory factors and pro-inflammatory factors before and after CPB and the correlation with CPB time were analyzed. Results:There were 65 boys and 59 girls with a body weight of(10.69±8.18)kg and a median age of 317(141, 975)d.After CPB, WBC(×10 9/L)(13.47 vs.8.6), NLR(4.93 vs.0.55), and CRP(mg/L)(81.35 vs.0.8) were significantly higher than those before operation( P<0.001). IL-6(pg/mL)(135.69 vs.6.86), IL-8(pg/mL)(33.33 vs.14.95), and IL-10(pg/mL)(6.05 vs.2.44)were significantly higher than those before operation( P<0.001). Compared with CPB<120 min group, IL-6(pg/mL)(211.88 vs.119.47), IL-8(pg/mL)(71.67 vs. 25.39), and IL-10(pg/mL)(7.69 vs. 4.92)in CPB≥120 min group significantly increased( P<0.001). CRP was negatively correlated with CPB time( r=-0.204, P=0.025), while IL-6( r=0.254, P=0.005), IL-8( r=0.358, P=0.001), IL-10( r=0.198, P=0.03) were positively correlated with CPB time.Twelve children(9.7%)had obvious SIRS, and four cases(3.2%)had early CARS.The mortality of CARS group was significantly higher than that of SIRS group and the control group( P=0.011). Conclusion:Il-6 , IL-8, and IL-10 are significantly increased after CPB in children with congenital heart disease.With the increase of CPB time, IL-6 and IL-8 increase significantly, and the correlation between IL-8 and CPB time is the strongest.Although the proportion of children with early postoperative CARS is small, the mortality is high, which indicates clinical surveillance and treatment need to be strengthened for anti-inflammatory response.

Family Function, Loneliness, Emotion Regulation, and Hope in Secondary Vocational School Students: A Moderated Mediation Model.

This study explored family function as a key factor of loneliness, hope, and emotion related to secondary vocational school students during the novel coronavirus pandemic. Chinese versions of the Olson Family Function Scale, Russell Loneliness Scale, Snyder Hope Scale, and Gross Emotion Regulation Scale were completed by 5,138 participants. Guardian type significantly predicted family function and loneliness. Family function significantly and positively correlated with loneliness. The relationship between family function and loneliness was mediated by hope, and expressive suppression concurrently moderated the relationship between hope and loneliness. Our study offers meaningful insights into the family function of Chinese secondary vocational school students. The findings supported a moderated mediation model that exemplifies the relationship between family function, loneliness, expressive suppression, and hope. Although the results showed that high hope mediated lower family functioning and loneliness, low expressive suppression led to intense loneliness immediately. This study confirms that emotional strategy is important and associated with mental health. It also suggests that schools should pay more attention to students' emotion regulation and help them rebuild hope or appropriate cognition to relieve loneliness during crisis events.

Network Meta-analysis of effect of traditional Chinese medicine injections in adjuvant treatment of mycoplasma pneumonia in children / 中国中药杂志

To assess the clinical efficacy of traditional Chinese medicine injection for adjuvant treatment of mycoplasma pneumoniae pneumonia in children by network Meta-analysis method. We retrieved CNKI, WanFang, CBM, Cochrane Library, PubMed from the establishment to September 2018. Two reviewers independently screened out literatures, extracted data and assessed the methodological quality of included studies. The data were analyzed by Stata 13.0 software. Totally 89 RCTs were included, involving 8 kinds of traditional Chinese medical injections and 8 936 patients. According to the results of network Meta-analysis, the order by the total effective rate from high to low was Huangqi Injection>Xiyanping Injection>Tanreqing Injection>Compound Danshen Injection>Reduning Injection>Yanhuning Injection>Qingkailing Injection>Xixinnao Injection; the order by cooling time from high to low was Reduning Injection> Yanhuning Injection>Qingkailing Injection>Tanreqing Injection>Huangqi Injection>Xiyanping Injection>Xiexinnao Injection>Compound Danshen Injection; the order by the cough disappeared time from high to low was Compound Danshen Injection>Qingkailing Injection>Xiyanping Injection>Huangqi Injection>Yanhuning Injection>Reduning Injection>Tanreqing Injection>Xixinnao Injection; the order by the rales disappearing time from high to low was Qingkailing Injection>Yanhuning Injection>Reduning Injection>Huangqi Injection>Tanreqing Injection>Xiyanping Injection>Xixinnao Injection. The results show that traditional Chinese medicine injection has a significant clinical efficacy in the adjuvant treatment of various symptoms of mycoplasma pneumoniae pneumonia in children. Due to the small sample size, more studies are required to verify the strength of evidence.

Antiviral effects of Bovine antimicrobial peptide against TGEV in vivo and in vitro

Background@#In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production.Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. @*Objectives@#This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. @*Methods@#The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID50 ). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. @*Results@#The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 µg/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log10 TCID50 of 62.5 µg/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 µg/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. @*Conclusions@#APB-13 exhibited good antiviral effects on TGEV invivo and in vitro.

Analysis of Selective Serotonin Reuptake Inhibitors Associated with QT Prolongation and Torsade de Pointes Adverse Events Based on Data Mining Methods / 中国药学杂志

OBJECTIVE: To excavate and evaluate the risk signals of QT prolongation and torsade de pointes(TdP) induced by selective serotonin reuptake inhibitors(SSRIs), provide references for clinical use. METHODS: Data from FDA adverse event reporting system (FAERS, from January 2004 through June 2018) were analyzed for each SSRIs, including fluoxetine, sertraline, citalopram, escitalopram, paroxetine, and fluvoxamine. When QT prolongation and TdP cases were identified using preferred terms (PT) and standardised MedDRA queries (SMQ), three different data mining algorithms were used to detect signalsreporting odds ratio (ROR), medicines and healthcare products regulatory agency (MHRA), and bayesian confidence popagation neural network (BCPNN), if all the three algorithms were positive, suggesting the generation of signals. RESULTS: A total of 3 912 reports of QT prolongation and TdP associated with SSRIs were retrieved through the SMQ. Among which, more females than males(2 349 vs. 1 150), mainly aged 18-44 and 45-64 years, and 90.64% were serious adverse events. The signals were found for fluoxetine, sertraline, citalopram, escitalopram, paroxetine and fluvoxamine at the SMQ level, the RORs (95%CI) were 5.25(4.79-5.76), 2.08(1.79-2.27), 2.86(6.32-7.44), 3.41(3.03-3.84), 2.09(1.84-2.37) and 10.44(8.17-13.33) respectively; the PRRs (X2) were 5.20(1 494.43), 2.01(140.41), 6.77(2 911.71), 3.93(462.34), 2.09(136.58) and 10.21(538.26) respectively; the Ics (IC-2SD) were 2.15(2.12), 1.54(1.52), 2.67(2.65), 2.34(2.31) 1.14(1.12) and 3.16(3.10) respectively. Analysis of the PT included in the SMQ for TdP/QT prolongation, except paroxetine was only detected electrocardiogram QT prolonged signal, all the other SSRIs were detected electrocardiogram QT prolonged and TdP signals. CONCLUSION: QT prolongation may be a SSRIs class effect, but TdP just for fluoxetine, sertraline, citalopram, escitalopram and fluvoxamine. Clinical staff should pay more attention to the differences in adverse drug reaction related to SSRIs, and take pertinence measure to prevent.

Chemical Constituents of Agricultural Residues Producing from 4 Kinds of Gramineous Crops and Their Pharmacological Effects / 中国实验方剂学杂志

Gramineous crops occupy a remarkable proportion of grain crops in the word,and wheat,rice and corn account for more than 80%of the world's food crops. Agricultural residues bring tremendous pressure on the environment,and inefficient development of resources has caused huge waste of resources. At present,the research on agricultural residues mainly focuses on energy,fertilizer,feed and materialization. However, there are still a lot of resources that have not been rationally utilized. The author has found that in recent years,the medicinal research on gramineous crop waste has focused on four varieties-rice,corn,wheat and sugar cane,and their waste rice bran,rice husk,rice straw,corn stigma,corn bract,wheat bran,sugar cane leaf,sugar cane skin. The compounds isolated and identified from agricultural residues include phenylpropanoids,flavonoids,steroids and their glycosides,organic acids and their esters,volatile oil and saccharides. Studies have shown that agricultural residues from gramineous crops have pharmacological activities, such as anti-oxidation,hypolipidemia,hypoglycemia,anti-inflammation,anti-tumor,anti-cardiovascular disease,anti-liver and kidney damage. This paper is a systematic review of the chemical composition and pharmacological effects of agricultural residues from the major gramineous crops,so as to provide useful information for further research and development of agricultural residues.

Clinical Efficacy of Xiaoai Jiedu Formula Combined with FEC Chemotherapy on Breast Cancer Patients / 中国实验方剂学杂志

Objective:To observe effect of Xiaoai Jiedu formula combined with fluorouracil (5-FU)+epirubicin (EPI)+cyclophosphamide (CTX) (FEC) chemotherapy regimen on immune function, tumor index, traditional Chinese medicine (TCM) symptom scale score and adverse reactions of patients with breast cancer. Method:A total of 60 patients with breast cancer were randomly divided into study group and control group. FEC (CTX 0.6 g·m-2,d1,EPI 100 mg·m-2,d1,5-FU 0.5 g·m-2,d1) regimen was used in study group, and Xiaoai Jiedu formula recipe was used for two consecutive cycles. FEC regimen was used in control group only. After 2 cycles, immunological changes, tumor index, TCM symptom score and adverse reactions were analyzed and compared between two groups. Result:There was not significant difference in immune indexes between two groups before treatment, but statistically significant differences after treatment (PPPPConclusion:Xiaoai Jiedu formula combined with FEC chemotherapy can improve clinical efficacy and alleviate clinical symptoms of patients.

Potential Value of Cancer-Associated Fibroblasts in Hematological Malignancies--Review / 中国实验血液学杂志

Cancer-associated fibroblasts (CAF), as one of the most important components of tumor microenvironment, which plays important role in tumorigenesis, development, infiltration and metastasis of cancers. In a variety of solid tumors, CAF can even determine the fate of tumor cells. In view of its pivotal role in promoting tumor progression, CAF has recently become a therapeutic target for a variety of tumors. However, there are a few studies on CAF in hematological malignancies. Recent studies have found that the resistance, relapse of AML, MM, CLL and myelofibrosis of MPN closely relate with CAF, so targeting CAF can effectively enhance the killing effect of chemotherapy drugs on tumor cells, thus improve the efficacy, CAF is expected to become a new target for the treatment of hematological malignancies. This review summarizes recent advances in cancer-associated fibroblasts in hematological malignancies.

Techniques and experience which improve the surgical outcomes of critical and complex congenital heart disease in neonates / 中华外科杂志

Objective@#To discuss the experience of early surgical intervention to critical and complex congenital heart diseases (CHD) in neonates.@*Methods@#A retrospective study of clinical records of patients with congenital heart diseases underwent surgical intervention in one single pediatric cardiac center was performed. From January 2009 to December 2017, 841 critical and complex CHD neonates were identified at Department of Cardiovascular Surgery, Children′s Hospital of Fudan University, of which 6.5% were premature. There were 557 male and 284 female patients. The age was (11±9) days, ranging from 0 to 28 days, M(QR): 8(17) days. The weight was (3.26±0.57) kg, ranging from 1.9 to 5.0 kg, M(QR): 3.3(0.7) kg. There were 189 patients associated with other malformation besides CHD. Before surgery, 13.6% (114/841) patients were on ventilation, and severe acidosis was addressed in 136 patients. All patients underwent surgical interventions, including surgical procedures and hybrid procedures.@*Results@#Emergency surgeries were performed in 35 patients. One hundred patients had palliative procedures, other 633 patients had complete repair, while the rest 108 patients underwent hybrid procedures. The in-hospital mortality was 4.0% (34/841). The ventilation time was 1.5 to 1 130.0 hours, M(QR): 94 (116) hours, with the rate of reintubation 3.3% (27/807). The ICU stay time was 1 to 195 days, M(QR): 14 (15) days.@*Conclusion@#The improvement on early screening and diagnosing, referral system, multidisciplinary cooperation and hybrid invention skills together contributed to better outcomes of critical and complex congenital heart diseases in neonates.

Research Progress of Histone Modification in B Cell Lymphoma Pathogenesis-Review / 中国实验血液学杂志

It has been increasingly recognized that the pathogenesis of B-cell lymphoma closely relates to the epigenetic disregulations. Epigenetics is a subdiscipline, which means heritable changes in gene expressions without alterations in the DNA sequence, and the DNA methylation, histone modification and miRNA maily were involved. Histone modification is the most important epigenetic modification, the researches showed that the aberrant histone modification is the important pathogenesis in B-cell lymphoma, especially the aberrant histone methylation and acetylation. In the meantime, the tumor can be treated by changing the epigenetic modification, which become a research hotpoint. This review summarizes the pathogenesis of B cell lymphoma and discusses the epigenetic treatment of B cell lymphoma mainly in terms of histone modification regulation for B cell development in the germinal center and mutation of histone madification enzymes.

Preparation of Plasma Membrane Vesicles from Bone Marrow Mesenchymal Stem Cells for Potential Cytoplasm Replacement Therapy.

We have previously reported on the generation of plasma membrane vesicles (PMVs) through the mechanical extrusion of mammalian cells. The fusion of PMVs with mitochondrial deficient Rho0 cells restored mitotic activity under normal culture conditions. Atherosclerosis, type 2 diabetes, Alzheimer's disease, and cancer are age-related diseases that have been reported to be associated with multiple mechanical and functional defects in the cytosol and organelles of a variety of cell types. Bone marrow mesenchymal stem cells (BMSCs) represent a unique cell population from the bone marrow that possess self-renewal capabilities while maintaining their multipotency. The supplementation of senescence cells with young cytoplasm from autologous BMSCs via the fusion of PMVs provides a promising approach to ameliorate or even reverse age-associated phenotypes. This protocol describes how to prepare PMVs from BMSCs via extrusion through a polycarbonate membrane with 3 µm pores, determine the existence of mitochondria and examine the maintenance of membrane potential within PMVs using a confocal microscope, concentrate PMVs by centrifugation, and carry out the in vivo injection of PMVs into the gastrocnemius muscle of mice.

VSV-G Viral Envelope Glycoprotein Prepared from Pichia pastoris Enhances Transfection of DNA into Animal Cells.

Vesicular stomatitis virus G glycoprotein (VSV-G) has been widely used for pseudotyping retroviral, lentiviral, and artificial viral vectors. The objective of this study was to establish a potential approach for large-scale production of VSV-G. To this end, VSV-G was cloned with an N-terminal His-tag into Pichia pastoris expression vector pPIC3.5K. Three clones (Muts) containing the VSV-G expression cassette were identified by PCR. All clones proliferated normally in expansion medium, whereas the proliferation was reduced significantly under induction conditions. VSV-G protein was detected in cell lysates by western blot analysis, and the highest expression level was observed at 96 h post induction. VSV-G could also be obtained from the condition medium of yeast protoplasts. Furthermore, VSV-G could be incorporated into Ad293 cells and was able to induce cell fusion, leading to the transfer of cytoplasmic protein. Finally, VSV-G-mediated DNA transfection was assayed by flow cytometry and luciferase measurement. Incubation of VSV-G lysate with the pGL3-control DNA complex increased the luciferase activity in Ad293 and HeLa cells by about 3-fold. Likewise, incubation of VSV-G lysate with the pCMV-DsRed DNA complex improved the transfection efficiency into Ad293 by 10% and into HeLa cells by about 1-fold. In conclusion, these results demonstrate that VSV-G could be produced from P. pastoris with biofunctionalities, demonstrating that large-scale production of the viral glycoprotein is feasible.

Incorporation of Viral Glycoprotein VSV-G Improves the Delivery of DNA by Erythrocyte Ghost into Cells Refractory to Conventional Transfection.

The objective of this study was to formulate a novel gene delivery system based on the erythrocyte ghost (EG) integrated with fusogenic viral glycoprotein vesicular stomatitis virus glycoprotein G (VSV-G). VSV-G proteins were harvested as condition medium of Ad293 cells carrying a VSV-G transgene and then incorporated into EG. Plasmid DNA was condensed by various transfection reagents. A luciferase expression construct (pGL3-control) and a DsRed expression cassette (pCMV-DsRed) were used to evaluate the delivery efficiency of DNA/EG/VSV-G complexes. VSV-G proteins could be incorporated into EG in static incubation under acidic conditions as evidenced by the Western blot analysis. Condensed plasmid DNA was bound mostly to the outer surface of EG, which could be detected by electromicroscopy and measured by electrophoresis. EG/VSV-G complexes stimulated the delivery of pGL3-control into Ad293 cells significantly with the luciferase activity increased about 4-fold as compared to that of the control. The delivery of pCMV-DsRed was also enhanced with the percentage of DsRed-positive Ad293 cells increased from 55 % to about 80 %. Moreover, the transfection efficiency in 3T3, HeLa, INS-1, and bone marrow stem cell (BMSC) cells increased about 2-3-fold. Finally, confocal microscopy analysis showed that incorporation of VSV-G significantly enhanced the endocytosis of EG into target cells. In the present study, a novel type of non-viral DNA delivery vehicle consisting of EG and fusogenic VSV-G proteins was formulated, which showed superior transfection efficiency even in cells resistant to classical transfection.

Clinical Observation of Kouyanqing Granules in the Treatment of Chronic Periodontitis / 中国药房

OBJECTIVE:To observe therapeutic efficacy and safety of Kouyanqing granules for chronic periodontitis and it effects on inflammatory cytokine. METHODS:110 patients with chronic periodontitis were divided into control group(55 cases)and observa-tion group(55 cases). Control group accepted the periodontal basic treatment as oral hygiene instruction,supragingival scaling,subgin-gival scaling and root planning. Observation group was additionally given Kouyanqing granules 20 g orally,twice a day,on the basis of control group. Both groups were treated for 4 weeks. Clinical efficacies of 2 groups were observed as well as the levels of plaque in-dex(PLI),gingival index(GI),probing depth(PD),clinical attachment level(CAL),IL-17,IL-21,and the occurrence of ADR. RE-SULTS:Both groups completed the treatment,and no patient lost to follow up. Total response rate of observation group was significant-ly higher than that of control group,with statistical significance(P0.05). CONCLUSIONS:Based on routine treatment,Kouyanqing granules show significant therapeutic efficacy for chronic periodontitis, can significantly improve periodontal symptom and reduce inflammatory factor level,but do not increase the incidence of ADR.

Response of human triple-negative breast cancer to paclitaxel after vascu-lar normalization in nude mice / 中国病理生理杂志

AIM:To explore whether there is synergistic effect of recombinant human endostatin ( rh-Endo ) and paclitaxel (Pac) in the time window of vascular normalization and the role of magnetic resonance imaging (MRI) in early assessment of chemotherapy by observing the response of human triple -negative breast cancer ( TNBC) to Pac after vascular normalization in nude mice .METHODS:The human TNBC MDA-MB-231 cells were planted in the subcutaneous region of right lower abdomen of BALB/c-nu female nude mice .These nude mice were randomly divided into 4 groups (n=7).rh-Endo was given for 17 consecutive days in rh-Endo group and rh-Endo+Pac group.Pac was given on the 6th and 12th days in Pac group and rh-Endo+Pac group.The dosage of both drugs was 10 mg· kg-1· d-1(ip).On the day before the treatment and the 5th, 11th and 17th days after treatment, all the transplanted tumors were examined by MRI . All the mice were killed by cervical dislocation and their transplanted tumors were taken down for examinations after the last MRI on the 17th day.The changes of pathology, immunohistochemisty, microvessel density (MVD) and Ki67 expression were measured.RESULTS:On the 17th day, the volume of transplanted tumor in rh-Endo+Pac group was smaller than that in model group and rh-Endo group ( P<0.05 ) , and no difference between rh-Endo+Pac group and Pac group was found.On the 17th day, the tumor inhibitory rates in rh-Endo group, Pac group and rh-Endo+Pac group were 14.61%, 39.08%and 54.79%, respectively.The slow diffusion coefficient in Pac group was increased compared with model group , while it was decreased compared with rh-Endo+Pac group (P<0.05).No distant metastatic lesion in the tumor-bearing mice was observed .The necrotic rates in rh-Endo+Pac group and Pac group were higher than those in model group and rh-Endo group.The MVD in model group was higher than that in the other 3 groups.The MVD in rh-Endo+Pac group was decreased compared with Pac group and rh-Endo group .The Ki67 level in rh-Endo+Pac group was decreased compared with rh-Endo group , and no difference between rh-Endo+Pac group and Pac group was detected .CONCLUSION:In the time window of vascular normalization , the combination of Pac and rh-Endo has a significant antitumor effect on TNBC , but this study did not observe a significant synergistic effect of the 2 drugs.The change of slow diffusion coefficient can predict the therapeutic effect in advance .

Response of human triple-negative breast cancer to paclitaxel after vascu-lar normalization in nude mice / 中国病理生理杂志

AIM:To explore whether there is synergistic effect of recombinant human endostatin ( rh-Endo ) and paclitaxel (Pac) in the time window of vascular normalization and the role of magnetic resonance imaging (MRI) in early assessment of chemotherapy by observing the response of human triple -negative breast cancer ( TNBC) to Pac after vascular normalization in nude mice .METHODS:The human TNBC MDA-MB-231 cells were planted in the subcutaneous region of right lower abdomen of BALB/c-nu female nude mice .These nude mice were randomly divided into 4 groups (n=7).rh-Endo was given for 17 consecutive days in rh-Endo group and rh-Endo+Pac group.Pac was given on the 6th and 12th days in Pac group and rh-Endo+Pac group.The dosage of both drugs was 10 mg· kg-1· d-1(ip).On the day before the treatment and the 5th, 11th and 17th days after treatment, all the transplanted tumors were examined by MRI . All the mice were killed by cervical dislocation and their transplanted tumors were taken down for examinations after the last MRI on the 17th day.The changes of pathology, immunohistochemisty, microvessel density (MVD) and Ki67 expression were measured.RESULTS:On the 17th day, the volume of transplanted tumor in rh-Endo+Pac group was smaller than that in model group and rh-Endo group ( P<0.05 ) , and no difference between rh-Endo+Pac group and Pac group was found.On the 17th day, the tumor inhibitory rates in rh-Endo group, Pac group and rh-Endo+Pac group were 14.61%, 39.08%and 54.79%, respectively.The slow diffusion coefficient in Pac group was increased compared with model group , while it was decreased compared with rh-Endo+Pac group (P<0.05).No distant metastatic lesion in the tumor-bearing mice was observed .The necrotic rates in rh-Endo+Pac group and Pac group were higher than those in model group and rh-Endo group.The MVD in model group was higher than that in the other 3 groups.The MVD in rh-Endo+Pac group was decreased compared with Pac group and rh-Endo group .The Ki67 level in rh-Endo+Pac group was decreased compared with rh-Endo group , and no difference between rh-Endo+Pac group and Pac group was detected .CONCLUSION:In the time window of vascular normalization , the combination of Pac and rh-Endo has a significant antitumor effect on TNBC , but this study did not observe a significant synergistic effect of the 2 drugs.The change of slow diffusion coefficient can predict the therapeutic effect in advance .

Expression of Proteus mirabilis polyphosphate kinase and preparation of its polyclonal antibodies / 南方医科大学学报

<p><b>OBJECTIVE</b>To express and purify polyphosphate kinase (PPK) from Proteus mirabilis and prepare the polyclonal antibody against PPK.</p><p><b>METHODS</b>The antigenicity and hydrophobicity of PPK were analyzed using software. The N-terminal conservative sequence containing 309 amino acids was selected as the target peptide, and its corresponding gene sequence with modification based on prokaryotic cells-preferred codon was synthesized and inserted into plasmid pET28b(+). The constructed recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and induced with IPTG. The expressed fusion protein was purified using Ni-affinity chromatography. The purified protein was injected along with adjuvant in rabbits to prepare the polyclonal antibodies against PPK.</p><p><b>RESULTS AND CONCLUSION</b>PPK fusion protein expressed by E. coli was purified successfully using Ni-affinity chromatography. ELISA result demonstrated that the harvested rabbit anti-sera against PPK had a high titer of 1:512 000, and Western blotting showed a good specificity of the antibody, which can be used further study of the role of PPK in the pathogenesis of Proteus mirabilis infection.</p>

Establishing the Ion Torrent PGM Sequencing Methods for the Clinical Diagnosis of MPN Patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and relibility of rapidly and accurately acquiring the informations of gene mutations in MPN patients by using self-designed custom MPN mutation-related multipe-PCR primer kit and next generation Ion Torrent PGM sequencing platform.</p><p><b>METHODS</b>The bone marrow samples of 10 MPN patients with JAK2V617F and/or CALR, Phconfirmed by sanger sequencing method were collected and were re-detected by using next generation Ion Torrent PGM sequencing method, then the consistence of results of above-mentioned 2 kinds of detection methods was compared.</p><p><b>RESULTS</b>In terms of JAK2V617F, MPL and CALR mutations, the results of Ion Torrent PGM sequencing were complete consistent with results of Sanger sequencing, except 52 bp deletion of CALR gene, which conld not be detected by next generation Ion Torrent PGM sequencing method in all bone marrow samples.</p><p><b>CONCLUSION</b>The detection of multiple gene mutations in MPN patients by Ion Torrent PGM sequencing platform is feasible and can meet the needs of clinical testing. This method can complete detection of all 23 mutetions within 1-2 days, moreover, possesses advantages of high sensitivity, specificity, rapidity, high throughput and low cost.</p>