Objective To construct and express fusion toxin human IL15 mutant/TNF? mutant (hIL15M/TNF?M) in order to develop a targeted toxin capable of eliminating abnormal cells expressing high levels of hIL15 receptors. Methods DNA fragments of truncated hIL15M and TNF?M coding gene were joined in frame and cloned into T7 expression vector pET16b. The expressed fusion protein was purified from E. coli . Cytotoxicity of hIL15M/TNF?M was measured by MTT assay. Results E. coli transformed with the recombinant plasmid expressed an extra band with the expected size of 30?10 3. MTT assay indicated that PHA activated T cells were 13 times more sensitive to the cytotoxic effect of hIL15M/TNF?M than to the resting T cells. Conclusion hIL15M/TNF?M has high toxic effect against cells expressing high levels of IL15 receptors, suggesting that hIL15M/TNF?M has potential therapeutic effect on T cell leukemia and other diseases related with improperly activated T cells.

Purpose:To study the expression of Epstein-Barr virus gene BARF1 in nasopharyngeal carc inoma cells, to uncover the mechanism and effects of Epstein-Barr virus in the pathogenesis of EBV related carcinomas, and to offer some experimental supports for the prevention and treatment of EBV related carcinomas Methods :After RNA extraction from nasopharyngeal carcinoma samples, RT-PCR was con ducted to amplify the EBV gene EB virus associated nuclear antigen-1 (EBNA1) an d the new gene Bam HI A open reading frame 1 (BARF1) Then the PCR products wer e electrophoresed by 2% agarose gel and the product bands were detected and phot ographed Results:Of the 13 samples, 11 were qualified for the RNA qualit y And all of the 11 NPC cases were EBNA1 positive, showing the EB virus infect ion in the carcinoma cells 9 of the 11 cases were BARF1 positive, the positive rate was nearly 82% Conclusions:The EB virus gene BARF1 was highly expressed in the NPC samples This implies t hat apart from the latent membrane protein 1 (LMP1), BARF1 may play an important role in the pathogenesis of NPC The exact mechanisms of the BARF1 effects on the pathogenesis of NPC need to be studied further

IrritabIe boweI syndrome( IBS)is caused by muItipIe factors,and the pathogenesis has not yet been cIarified. RecentIy,the roIe of brain-gut axis based bio-psycho-sociaI medicaI modeI in the pathogenesis of IBS has been wideIy accepted. Brain-gut axis forms a nerve-endocrine-immune network mediating bidirectionaI reguIation pathway,which pIays an important roIe in maintaining homeostasis between centraI nervous system and gut. Brain-gut axis abnormaIities may Iead to imbaIance of the homeostasis and subsequentIy induce the occurrence of IBS. This articIe reviewed the advances in study on brain-gut axis dysfunction in pathogenesis of IBS.

Objective To study the expression changes of?-catenin in the hair follicle before and after GasderminA3 gene mutation.Methods Using SP immunohistochemical staining,RT-PCR,Western blotting to detect the expression of?-catenin in the hair follicle in GSDMa3 mutant and C57BL/6(B6) mice on postnatal 11(anagen) ,16(early catagen) ,18(late catagen) and 24 d(telogen) respectively.Results The expression of?-catenin is gradually weakened from 11 d to 24 d in GSDMa3 mutation mice and B6 mice,but stronger expression was found in GSDMa3 mutation mice than in the B6 mice at different time points.In anagen,?-catenin was expressed in the inner and outer root sheath and hair matrix cells,with stronger expression in GSDMa3 mutant mice than in B6 mice.In catagen,?-catenin was mainly expressed in the outer root sheath and hair matrix cells,with more stronger expression in GSDMa3 mutant mice than in B6 mice.In telogen,?-catenin was expressed in the outer root sheath cells in the mutant mice while little in the hair follicle in B6 mice.Conclusion GSDMa3 gene affects the hair follicle growth and development,probably through regulating the expression of?-catenin.

Objective To investigate the predialysis education on dialysis timing of patients with end-stage renal disease (ESRD). Methods 129 patients with chronic renal failure (CRF) and serum creatinine(Scr)＞442μmol/L, endogenous creatinine clearance rate (Ccr)＜20 ml/min were randomly divided into the experimental group(70 cases) and the control group(59 cases), the experimental group received predialysis education, the control group received routine care.The basic situation, dialysis timing, depression and quality of life on the 6th,12th months after dialysis were assessed. Results There was no significant difference in age, sex, education background, health care payment manners, employment, serum creatinine, endogenous creatinine clearance rate before education.For the timing of dialysis, Ccr for the experimental group was (9.49 ± 0.77)ml/min, Ccr for the control group was (4.54 ±1.79) ml/min,the difference was significant, depression and quality of life between two groups after 6,12 months of dialysis was also statistically different. Conclusions Strengthening predialysis education is conducive to a timely start of dialysis and can effectively improve the quality of life in patients with ESRD.

Objective To investigate the role of N-methyl-D-aspartate receptor one(NMDA-R1 )in visceral hypersensitivity forming of Sprague-Dawley rat model induced by transient Trichinella spiralis intestinal infection.Methods Forty male Sprague-Dawley rats were divided into healthy control group,visceral hypersensitivity group,0.9％ NaCl group and MK-801 group.The rats in visceral hypersensitivity group,0.9％NaCl group and MK-801 group were given Trichinella spiralis to establish infection model.After eight weeks,the rats in 0.9％NaCl group and MK-801 group were given 0.9 ％ NaCl and MK-801 respectively.The abdominal muscular electrical activity of each group was observed under 20,40,60 and 80 mm Hg (1 mm Hg=0.133 kPa) colorectal distension pressure.After abdominal muscular electrical activity recorded,rats were sacrificed and the colon specimens were collected.The expression of NMDA-R1 at protein level was detected by Western blot.The data were analyzed by LSD test and Bivariate correlation.Results There was significant correlation between areas under curve (AUC) of abdominal muscular electrical activity electromyography and distension pressure in healthy control group,visceral hypersensitivity group,0.9％ NaCl group and MK-801 group (r=0.823,0.618,0.913,0.889 respectively,all P＜0.01).Under 20,40,60,80 mm Hg distension pressure,there was significant difference in AUC between MK-801 group and visceral hypersensitivity group (LSD test,all P＜0.05).The AUC of MK-801 group was lower than that of healthy control group,but there was significant difference (LSD test,P =0.029) only when distension pressure was at 80 mm Hg.The integral optical density value of NMDA-R1 protein of MK-801 group (1.238 ±0.210) was significantly lower than that of visceral hypersensitivity group (2.231±0.450) and 0.9％NaCl group (2.104±0.220) (LSD teat,P=0.025,0.046).The JA value of NMDA-R1 protein of visceral hypersensitivity group and 0.9％NaCl group was significantly higher than that of healthy control group (LSD test,P =0.007,0.014).Conclusion The visceral hypersensitivity forming was correlated with the high expression of NMDA-R1,and it could be adjusted by inhibiting NMDA-R1 expression in enteric nervous system.

AIM:To characterize the inclusion complex of volatile oil of Rhodiola crenulata/?-cyclodextrin. METHODS: Some analytical methods,such as DTA,IR,GC,TLC were apllied to the investigation before and after the inclusion. RESULTS: The difference between the inclusion and physical mixture in differential thermal(analysis,) infrared spectra,gas chromatogram and thin layer chromatogram. CONCLUSION: The inclusion of Rhodiola crenulata volatile oil has the characteristic of intramolecular inclusion.

Objective:To evaluate early corneal biomechanical change, safety and effectiveness after small-incision lenticule extraction combined with ultraviolet A corneal collagen cross-linking (SMILE-CXL) for myopic eyes and compare with SMILE.Methods:A non-randomized controlled clinical study was performed.Forty-four myopic eyes of 25 patients were included in Hankou Aier Eye Hospital from December 2017 to July 2018.SMILE-CXL or SMILE was carried out for 22 eyes of 12 patients or 22 eyes of 13 patients, respectively, based on the normal posterior surface manifestation of Belin by Pentacam tomography or not.The posterior surface manifestation of Belin was normal in the SMILE group, and the posterior surface of Belin was yellow or red with Corvis biomechanical index and tomographic biomechanical index<0.3 (except keratoconus) in the SMILE-CXL group.The refractive diopter of the eyes was detected before and 6 months after surgery, including spherical diopter, cylindrical diopter and spherical equivalent (SE) with a comprehensive refractometer.The corneal biomechanical parameters of the eyes were detected before and 6 months after surgery with a Scheimpflug-based dynamic tonometry (Corvis ST). The safety index, a ratio of postoperative mean best corrected visual acuity (BCVA) to preoperative mean BCVA, and efficacy index, a ratio of postoperative uncorrected visual acuity (UCVA) to preoperative BCVA, were compared between SMILE-CXL group and SMILE group.BCVA and UCVA were examined using standard chart and converted to logarithm of the minimal angle of resolution (LogMAR) units.This protocol complied with the Declaration of Helsinki and was approved by an Ethics Committee of Hankou Aier Eye Hospital (No.WHS2017052701). Written informed consent was obtained from each patient prior to entering into the cohort.Results:The postoperative UCVA of the eyes in both groups was more than 1.0.There were no significant differences in safety index, efficacy index and SE change between the SMILE-CXL group and SMILE group (all at P≥0.05). At 6 months after surgery, the values of second applanation time (A2T), time from the start until the highest concavity (HC-Time) and DA ratio 2 mm were significantly increased in comparison with before operation in both SMILE-CXL group and SMILE group (all at P<0.05). The changes of A2T, HC-Time and DA ratio 2 mm in the SMILE-CXL group were significantly lowered than those in the SMILE group at the sixth month after surgery ( P=0.001, 0.001, 0.036). The deformation amplitude, maximum corneal velocity during the first applanation (Vin), distance between both non-deformed peaks and integrated radius during the maximum depression were significantly increased, and the central curvature radius at highest concavity, cord length of first applanation, cord length of second applanation, Ambrósio's relational thickness horizontal, stiffness parameter applanation 1 and biomechanical intraocular pressure during the maximum depression were significantly lower at the sixth month after surgery than those before surgery (all at P<0.05). There were no significant differences in the parameters mentioned above between the two groups (all at P<0.05). Conclusions:Compared with SMILE alone, SMILE-CXL shows a comparable safety and efficacy, and better corneal biomechanical properties.

Objective: To explore the impact of arsenic on cholesterol efflux and the expression of ATP-binding cassette, sub-family A, member 1 ( ABCA1 ), ATP-binding cassette transporter G1 ( ABCG1 ), and scavenger receptor class B member I ( SRBI ) in macrophages, so as to provide the evidence for the mechanism of arsenic induced atherosclerosis. Methods: The human myeloid leukemia mononuclear cells ( THP-1 ), induced by phorbol myristate acetate, and mouse primary macrophages were treated with 0, 0.625, 1.25, 2.5 and 5 μmol/L NaAsO2 for 48 hours. Then the cells treated with 2.5 μmol/L NaAsO2 were changed to arsenic free mediums for 48 hours and collected every 12 hours to analyze the time effect of arsenic. The expression levels of ABCA1, ABCG1 and SRBI were determined by quantitative polymerase chain reaction and western blotting. Cholesterol efflux rates were measured by 3H isotope tracer. Results: Arsenic significantly down-regulated the expression levels of ABCA1 and ABCG1, and cholesterol efflux in a dose-dependent manner. The levels of ABCA1 mRNA decreased by 69% and 72%, the levels of ABCG1 mRNA decreased by 42% and 34%, and the rate of cholesterol efflux decreased by 55% and 59% in THP-1 and mouse primary macrophages cells treated with 5 μmol/L NaAsO2 ( all P<0.05 ). Arsenic had no significant effect on SRBI expression ( all P>0.05 ). Arsenic inhibited ABCA1 expression and cholesterol efflux in THP-1 in a time-dependent manner. Compared with cells before the exposure of arsenic, the level of ABCA1 mRNA and the rate of cholesterol efflux in THP-1 bottomed at 48 hours by 43% and 42%, and gradually recovered when arsenic was removed. Conclusions Arsenic inhibits cholesterol efflux by down-regulating the expression of ABCA1 and ABCG1 in macrophages.

Objective To determine the clinical value of laparoscopic right hepatectomy (LRH) carried out following a standardized technique.Methods The medical records of 15 patients who underwent LRH at the First Affiliated Hospital of Soochow University were retrospectively reviewed.The perioperative indicators which included the operation duration,blood loss,tumor diameter,hospitalization duration and postoperative complications were analyzed.Results There was no conversion to open surgery.The operation duration was (251.1 ± 73.3) min.The resected tumor diameter was (8.5 ± 4.2) cm,the blood loss was (550.8 ± 343.6) ml,and the hospitalization duration was (10.2 ± 3.7) days.There was one patient who developed postoperative bile leakage.There was no other complication and there was no perioperative death.Conclusions The standardized technique of LRH was easy and effective.This technique shortened the operation duration and improved patient safety.