[摘 要] 可变剪接是转录后水平的基因表达调控机制，也是导致真核生物转录组和蛋白质组多样性的重要途径。然而，可变剪接的异常是驱动肿瘤进展的重要推手。在肿瘤微环境中，肿瘤细胞、免疫细胞及肿瘤中其他类型细胞中mRNA的异常剪接，不仅参与塑造肿瘤细胞的恶性生物学行为和免疫逃逸，还促进支持肿瘤进展的免疫抑制性微环境的形成。靶向肿瘤相关剪接体组分、剪接调控因子、可变剪接产生的蛋白异构体和mRNA变异体，以及异常可变剪接产生的肿瘤新抗原已成为肿瘤治疗的新策略，已有基于可变剪接的肿瘤生物治疗项目进入到Ⅰ期临床研究阶段。基于可变剪接的肿瘤治疗面临安全性、长读测序和算法优化、核酸类药物递送等许多尚待解决的科学和技术问题，这些挑战的解决将为精准筛选肿瘤相关靶点和高免疫原性新抗原，突破传统疗法耐药瓶颈，增强免疫检查点阻断和CAR-T细胞等疗效提供新策略，开辟新领域。

Pancreatogenic diabetes is a type of diabetes secondary to pancreatic exocrine disease,and it was officially named by American Diabetes Association in 2014.Chronic pancreatitis and pancreatic cancer are the most common causes of pancreatogenic diabetes.The pathogenesis of this disease remains unclear,and there is still a lack of systematic treatment regimens,which leads to the extremely high misdiagnosis rate of pancreatogenic diabetes in China and globally.In addition,studies have shown that compared with patients with type 2 diabetes,patients with pancreatogenic diabetes tend to have higher risks of death and readmission,which brings great challenges to the health and clinical treatment of patients.Therefore,the comprehensive understanding and early accurate identification and diagnosis of pancreatogenic diabetes are of great significance in reducing the disability and mortality rates of this disease.This article elaborates on the possible pathogenesis of pancreatogenic diabetes.

Objective To determine the quantitative diagnostic criteria of phlegm-stasis interjunction syndrome of metabolic syndrome.Methods Based on the literature research,the Expert Consultation Questionnaire for MS Syndrome of Phlegm-stasis Interjunction based on Analytic Hierarchy Process was developed.Experts were invited to fill out the questionnaire,and the Diagnostic Criteria for MS Syndrome of Phlegm-stasis Interjunction(Draft)was constructed by using the expert group decision algorithm.The patients with MS were prospectively collected,and the diagnostic criteria for phlegm-stasis interjunction syndrome of MS were validated and revised by conditional probability conversion method and maximum likelihood discrimination,and the quantitative diagnostic criteria for phlegm-stasis interjunction syndrome of MS were finally formulated.Results The sensitivity,specificity and accuracy of the established quantitative diagnostic criteria for MS phlegm-stasis interjunction syndrome were 95.7%,84.0%,91.6%,positive likelihood ratio 5.98,negative likelihood ratio 19.46.Conclusion The quantitative diagnostic criteria for MS phlegm-stasis interjunction syndrome established based on literature research,expert consultation and diagnostic tests have good diagnostic efficacy.

BACKGROUND:Animal models of osteoporosis play an important role in the research of the pathogenesis, occurrence and development of osteoporosis, as wel as in the clinical diagnosis, prevention and treatment of osteoporosis. OBJECTIVE: To summarize and discuss the establishment and research ideas of osteoporosis models, explore the current situation and advance of osteoporosis models, compare the advantages and disadvantages of various methods, and provide evidence for clinical investigation. METHODS: A computer-based online search was conducted in SinoMed, VIP, Wanfang and PubMed databases by using the key words of “animal model, osteoporosis” from January 1969 to October 2015. The language was limited to both Chinese and English. Relevant articles were screened according to inclusion and exclusion criteria. The documents about the methods of osteoporosis model preparation, method improvement as wel as their advantage and disadvantage were summarized. RESULTS AND CONCLUSION:A total of 576 articles were included. Among them, articles published earlier, duplicated, and similarly were excluded, and 53 articles were finaly included. Various animal models of osteoporosis may only focus on the certain causes, certain stage, some of the main symptoms and some pathophysiological changes of disease. Accordingly, appropriate modeling methods and experimental animals should be selected based on research objective. Rat undergoing castration is the most commonly used model in the modeling of osteoporosis. Among drug methods for constructing osteoporosis model, glucocorticoids is the most commonly used one. Disuse method and nutritional method have limitations, and always combined with castration and drug methods. The effects of gene transfer, gene mutation and brain-derived model deserve

Objective To analyze non -small cell lung cancer patients with platinum -based chemotherapy excision repair cross -complementing 1 (ERCC1)and breast cancer susceptibility gene 1 (BRCA1 )gene polymor-phism,to examine the correlation of ERCC1 and BRCA1 gene polymorphism and non -small cell lung cancer platinum chemotherapy drugs sensitivity and chemotherapy prognosis.Methods 140 cases of non -small cell lung cancer were selected as subjects of this study.All patients were given platnum -based chemotherapy,peripheral blood ERCC1 and BRCA1 genes polymorphism were determined.The distribution of ERCC1 Asn118Asn genotype and BRCA1 Ser1613Gly genotype was observed.The relationship between different genotypes and the effect of chemotherapy and survival time after chemotherapy was compared.Results The proportions of ERCC1 Asn118Asn TT genotype,CT genotype and CC genotype were 5.7%,30.7% and 63.6%.The proportions of BRCA1 Ser1613Gly GG genotype,AG genotype and AA genotype were 8.6%,52.9% and 38.6%.In 140 patients,completely cured,partial response,stable disease and progressive disease patients were 0 case,33 cases,61 cases and 46 cases,the proportions were 0.0%, 23.6%,43.6% and 32.9%,the chemotherapy effective rate was 33.8%.ERCC1 Asn118Asn genotype was signifi-cantly correlated with the effect of non -small cell lung cancer chemotherapy (χ2 =4.416,P <0.05 ).BRCA1 Ser1613Gly genotype was significantly correlated with the effect of non -small cell lung cancer chemotherapy (χ2 =13.256,P <0.05).By Cox regression analysis and Log -rank test analysis,the average survival time of BRCA1 Ser1613Gly gene CC genotype non -small cell lung cancer after chemotherapy was longer than the CT +TT genotype (OR =2.946,χ2 =5.136,P <0.05).The average survival time of BRCA1 Ser1613Gly gene AA genotype non -small cell lung cancer after chemotherapy was shorter than the AG +GG genotype (OR =3.124,χ2 =5.136,P <0.05).Conclusion ERCC1 Asn118Asn genotype and BRCA1 Ser1613Gly genotype was significantly correlated with non -small cell lung cancer platinum -based chemosensitivity and chemotherapy prognosis.

Objective To investigate the effect of blood-bag placement mode in the refrigerator on the expression of adherent molecules in the preservation of human erythrocytes onboard the ship.Methods On the first day of deployment to sea, 4 blood samples were collected from 4 healthy volunteers.Erythrocyte suspension was equally aliquoted into four portions, one as the control group was placed vertically in the refrigerator of the hospital and the other three as the experiment group were preserved in the same refrigerator on-board the ship in 3 different placement modes ( the layer-upon-layer mode, the upright mode and the vertical suspension mode) .After completion of the training, the preserved samples were immediately sent to the laboratory for the detection of the expressions of CD47, CD58, and CD44.Results The expression levels of CD44 and CD47 in the erythrocytes of whole blood placed on the shipboard refrig-erator were significantly higher, as compared with those of the control group in the hospital.The expression levels of CD58 in the eryth-rocytes placed in the layer-upon-layer mode decreased significantly.Conclusion From the aspect of the expression of erythrocyte, the upright placement mode was most suitable for the preservation of blood in the shipboard refrigerator.

BACKGROUND: Some researches demonstrate that jog stress at fracture part can promote formation of callus, but molecular biological mechanism of jog stress on fracture healing is still unclear.OBJECTIVE:To study the effect of jog stress at fracture site on basic fibroblast growth factor(bFGF)at broken parts.DESIGN:Randomized controlled animal study.SETTING:Department of Orthopaedics,General Hospital of the Second Artilleryman of Chinese PLA;Departments of Orthopaedics and Pharmacy,the First Affiliated Hospital of General Hospital of Chinese PLA.MATERIALS:The experiment was carried out in the Animal Laboratory,the First Affiliated Hospital of General Hospital of Chinese PLA and the 8th Laboratory,Academy of Military Medical Sciences of Chinese PLA from March 2003 to April 2004.A total of 72 healthy purebred New Zealand rabbits,of general grade,5-6 months old,weighing 2.5-3.0 kg,were provided by Animal Center of Academy of Military Medical Sciences of Chinese PLA.All rabbits were randomly divided into jog group and fixation group with 36 rabbits in each group.And then,every 6 rabbits in both groups were respectively observed at 6 time points,including 7,14,21,28,42 and 56 days.METHODS:All animals were anesthetized with ketamine and sumian xinji.Fixation needle was rotated into 3,3.5,5.5 and 6 cm below platform of tibia,respectively,and then external fixators were installed.Chuck which was located at 1.5 cm from internal cortical bone and 4.5 cm below platform of tibia crossly cut off tibia.Fracture space was 2.0 mm and 2.5 mm in the fixation group and jog group,respectively.Animals in the fixation group were fixed with unilateral external fixator to dissect replace fracture part.In addition,animals in the jog group cut off their bone,fixed,and moved 0.5 mm to axis along central staff of external fixator.Animals were able to freely walk and external fixator was slightly shaken based on body mass of animal.Furthermore,all animals were sacrificed at 7,14,21,28,42 and 56 days after operation.According to the center of fracture part,samples in 1 cm length were cut,divided and fixed for 12 hours.Quantitative analysis of bFGF and colorized intensity were detected with immunohistochemical stain and JVC imaging analytic system at 7,14,21,28,42 and 56 days after operation.MAIN OUTCOME MEASURES:①Detecting colored intensity of bFGF;②quantitative analysis of bFGF.RESULTS:A total of 72 rabbits were involved in the final analysis bFGF could express in interstitial cells,vascular endothelial cell,osteoblast,chondroblast and osteocyte plasma.At 14,21 and 28 days after operation,positive indexes of bFGF protein in the jog group were 1.98±0.14,2.04±0.12 and 2.13±0.17,respectively,which were obviously higher than those in fixation group(1.59±0.14,1.68±0.15,1.63±0.27,P＜0.05).CONCLUSION:Jog stress can increase numbers of bFGF at fracture part and promote fracture healing.

Objective To study the effect of Vincristine-loaded intact human erythrocytes on tumor cell in vitro. Methods VCR was loaded in intact human erythrocytes using a modified hypotonic preswelling technique. The proliferation of K562 cells after incubation with VCR-loaded erythrocytes was detected by MTT. Cellular cycle and apoptosis of K562 cells were analyzed through FACS staining with PI/Rhodamin123, and morphology of apoptotic K562 cells and their nucleus were observed through staining with Hoechest33258. Results VCR-loaded erythrocytes could efficiently inhibit the proliferation of co-cultured K562 cells and induce the cells′ apoptosis in a dosage-dependent manner. Cellular cycle analysis demonstrated that proportion of K562 cells in G 0/G 1 decreased whereas in S+G 2/M increased after incubation with VCR-loaded erythrocytes. Such effects could also be displayed from alone VCR group, and significant change was observed in K562 cells co-cultured with unloaded erythrocytes (P

Objective To evaluation of the result of transplantation of BDNF gene transfected olfactory ensheathing cells (OECs) to repair spinal cord injury (SCI). Methods The rat with SCI were divided into non-OECs, OECs and transfected OECs groups. The evidences of anatomical regeneration across the transection site were demonstrated by both neuronal tracing and immunohistochemical methods. Results 12w after implantation the retrograde labeling study with flurogold showed abundant labeled neurons in the superior part of CNS. There were a lot of labeled neurons in the intermediate zone and dorsal column of lower thoracic spinal cord rostral to the transection site. Labeled neurons were also observed in the upper thoracic and cervical spinal cord, reticular nucleus and raphe nucleus of oblongata, as well as reticular nucleus and parabrachial nucleus of pons. The anterograde labeling of PHA-L showed that regenerated fibers of upper neurons overcame glial scar and regrew into the distal part of spinal cord through the grafts. Some labeled fibers extended long enough to the caudal gray matter of the transected site. Conclusion The implantation of OECs transfected by BDNF gene may benefit the survival and regeneration of the injured axons, and accelerate the repair of the injured spinal cord in a more efficient way than that with OECs alone.

Objective To investigate the expression of IL-8 and CXCR2 on keratinocytes from psoriatic lesions and their roles on clinical and pathologic manifestations. Methods The chemotaxis of psoriatic lesional keratinocytes was detected by micropore loculus test. The concentration of IL-8 was determined in the cultured supernatants of psoriatic keratinocytes by ELISA. The expression of CXCR2 on keratinocytes from affected skin was tested by flow cytometry. Results The chemotaxis for neutrophils by the cultured supernatants of psoriatic lesional keratinocytes was significantly stronger than that by controls. The concentration of IL-8 in the cultured supernatants of psoriatic lesional keratinocytes was also increased. The expression of CXCR2 on psoriatic keratinocytes was significantly increased. Conclusions The psoriatic epidermal hyperproliferation may be correlated with up regulation of IL-8 production and CXCR2 expression on psoriatic keratinocytes. At the same time, the psoriatic inflammation may be partly related to the increase of secretion of IL-8, which has chemotactic capacity, by keratinocytes. IL-8 and CXCR2 may be involved in the pathogenesis of psoriasis.