AIM:To investigate the effect of ankyrin repeat domain 49(ANKRD49)on epithelial-mesenchy-mal transition(EMT)in NCI-H1299 cells,and to explore its mechanism.METHODS:The ANKRD49 was over-ex-pressed in NCI-H1299 cells.The morphological changes of ANKRD49-overpressing NCI-H1299 cells were observed under microscope.The mRNA and protein expression levels of EMT-related markers[E-cadherin,transforming growth factor-β1(TGF-β1),vimentin and α-smooth muscle actin(α-SMA)]and EMT-related transcription factors(Snail1,Slug,Twist and ZEB1)were detected by RT-qPCR Western blot.Immunofluorescence staining was performed to observe the localiza-tion and expression of E-cadherin and vimentin in ANKRD49-overexpressing cells or control cells.Immunohistochemical method was performed to examine the levels of E-cadherin,α-SMA,Snail,Slug and ZEB1 in lung tissues of nude mice in-oculated with ANKRD49-overexpressing H1299 cells or control cells.RESULTS:Compared with the control group,the ANKRD49 overexpressing cells showed mesenchymal cell morphology(fusiform and less tight connections).RT-qPCR and Western blot results showed that the mRNA and protein levels of mesenchymal markers vimentin and α-SMA in ANKRD49 overexpressing cells were significantly higher than those in cells of control group,while the mRNA and protein levels of epithelial marker E-cadherin were lower than those in cells of control group.Compared with control group,the im-munofluorescence intensity of E-cadherin of H1299 cells decreased in after ANKRD49 overexpression,while that of vimen-tin increased significantly.Snail,Slug and ZEB1 expression were significantly elevated in ANKRD49 overexpressing cells compared with control group.The levels of E-cadherin in lung tissues of nude mice inoculated with ANKRD49-overexpressing H1299 cells declined,while the levels of α-SMA,Snail,Slug and ZEB1 increased compared with those in control mice.CONCLUSION:ANKRD49 promoted EMT of NCI-H1299 cells by increasing the expression of Snail1,Slug and ZEB1 and consequent downreguation of E-cadherin and upregulation of vimentin and α-SMA.

Abstract： Objective　To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods　In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results　The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions　In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.

OBJECTIVE To analyze the medication reconciliation for the inpatients in cardiovascular medicine department ，to provide reference for the establishment of working mode of clinical pharmacists in the department of cardiovascular medicine and to provide a basis for clinical pharmacists and community pharmacists developing pharmaceutical care for patients after transfering to community health center. METHODS From October 2020 to September 2021，newly admitted or newly transferred inpatients with chronic disease were selected from Shiyan People ’s Hospital of Shenzhen Bao ’an District. Medication reconciliation was conducted by clinical pharmacists after pharmaceutical consultation. According to the Pharmaceutical Care Network Europe （PCNE） classification system V 9.1，the existing drug-related problems （DRPs）were classified and summarized. The effectiveness and safety evaluation，medication education and other measures were provided ，and the acceptance of intervention was analyzed at the same time. RESULTS A total of 100 patients were included ，including 54 males and 46 females. The average age was （60.21±9.69） years，the average number of chronic diseases was （2.84±0.83），and the median number of drugs was 5.00. Among them ，110 treatment drug deviations were found in 74 patients，involving 10 categories and 61 drugs. Top three drugs in the list of accumulative drug deviation were cardiovascular system drugs （35 deviations），digestive medicine drugs （16 deviations）and endocrine system drugs （15 deviations）. The above treatment drug deviation may cause 122 DRPs， mainly “treatment effectiveness”problems（74 DRPs），and the causes were “inappropriate medication time ormedication interval ”（32 DRPs）， followed by “inappropriate drug combination ”（10 DRPs）. Interventions to DRPs mainly concentrated on patient level ，drug level （58）and doctor level （58），155 of which （84.70％）were fully accepted and implemented. CONCLUSIONS Some patients have a weak awareness of medications according to doctor ’s advice；drug reconciliation led by clinical pharma- cists at admission can fully understand the potential drug problems of patients ，and help doctors improve the drug compliance of patients and ensure their medication safety .

8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.
8-Hydroxy-2'-Deoxyguanosine ; Animals ; Antibodies, Monoclonal ; Gold ; Gold Colloid/chemistry* ; Metal Nanoparticles ; Mice ; Sensitivity and Specificity

Objective To evaluate the effect of an inverse planning simulated annealing (IPSA) in the treatment of cervical cancer with combined intracavitary and interstitial three-dimensional brachytherapy.Methods A total of 60 patients with locally advanced cervical cancer who received both external beam radiotherapy and combined intracavitary and interstitial brachytherapy in our hospital from October 2016 to July 2018 were enrolled.Patients were divided into four groups with 15 patients each according to the number of needles applied (1,2,3,and 4 needles,respectively).Dosimetric distributions were optimized with both Graphical optimization (GRO) and IPSA.Paired t-test was applied to compare the dosimetric differences between plans optimized with GRO and IPSA.Results The Dg0 and V100 of IPSA plans were higher than those of GRO (t=-4.742,-4.823,P＜0.05),while the conformity index (CI) and conformal index (COIN) were slightly lower than those of GRO plans (t=9.642,8.783,P＜0.05).No significant difference in the V150,V200,V300 between IPSA and GRO (P＞0.05) was observed.There was also no significant difference in the D2cm3 of bladder and rectum between IPSA and GRO (P＞0.05).The difference of Dg0 between IPSA and GRO was increased as the number of implanted needles increased,which increased from 4 cGy to 14 cGy as the number of needle increased from 1 to 4.The difference of V100 between GRO and IPSA was also increased as the number of needle increased.Conclusions In the treatment of cervical cancer with combined intracavitary and interstitial threedimensional brachytherapy,IPSA plan could improve the target coverage(D90,V100)without increasing the dose to the OARs and high dose region in the target compared with GRO.With the numbers of needles increased,the advantage of IPSA increased in terms of target coverage.

Objective: To investigate the clinical efficacy of subacromial anterolateral small incision approach with open reduction and internal fixation with proximal humeral locking system (PHILOS) for proximal humeral fractures. Methods: A retrospective case-control study was conducted to analyze the complete medical records of 76 patients with proximal humeral fractures admitted to the Department of Traumatic Surgery, Fujian Provincial Hospital from April, 2013 to December, 2017. There were 22 males and 54 females, aged from 25 to 89 years [(60.4±16.4)years]. All patients had closed fractures. According to Neer classification, there were 12 patients of type II, 46 patients of type Ⅲ, and 18 patients of type IV. The duration from injury to operation ranged from 1 to 10 days [(4.4±1.9)days]. All patients received treatment of open reduction fixation with PHILOS. Forty patients received subacromial anterolateral small incision approach with percutaneous interactive reduction and internal fixation of humerus head and humerus shaft, including four patients of Neer type II, 26 patients of type III, and 10 patients of type IV (minimally invasive group). Thirty-six patients received the deltopectoral approach with reduction and internal fixation, including eight patients of type II, 20 patients of type III, and 8 patients of type IV (conventional group). The total length of incision, the operation time, intraoperative blood loss, hospitalization time, review of the neck angle with X-ray 1 week after surgery, and the fracture healing after 6 months, the ipsilateral Neer shoulder joint function score at 6 months after surgery and complications were compared. Results: All patients were followed up for 3-12 months [(9.2±1.7)months], and there were six patients lost to the follow-up including four of the minimally invasive group and two of the conventional group. In the minimally invasive group and the conventional group, the total length of incision was 6.0(6.0, 6.8)cm and 11.5(10.0, 15.0)cm ( P<0.01), the operation time was (122.2±31.8)minutes and (136.9±36.6)minutes ( P>0.05), the intraoperative blood loss was 100(80, 150)ml and 175(100, 200)ml (P<0.01), the hospitalization time was (15.3±8.3)days and (16.2±5.1)days (P>0.05), the neck-shaft angle was (134.7±2.5)° and (134.6±2.6)°(P>0.05). A total of 70 patients obtained good bone healing. At 6 months after operation, the Neer shoulder function score of the affected side in the minimally invasive group was excellent in 30 patients, good in four patients, and fair in two patients, with the excellent and good rate of 96%, and for conventional group the score was excellent in 20 patients, good in eight patients, and fair in six patients, with the excellent and good rate of 84%(P<0.05). There was no complication in the minimally invasive group, but axillary nerve injury was found in one patient in the conventional group. Conclusion For the proximal humerus fracture, compared with the traditional deltopectoral approach, the subacromial anterior lateral small incision approach with percutaneous humeral head and humeral shaft reduction and interal fixation has the advantages of smaller incision, less bleeding and better functional recovery, which is a minimally invasive and effective treatment.

OBJECTIVETo investigate the pathogenic and clinical presentation and laboratory tests of bullous rash in hand, foot and mouth disease (HFMD) in Xi'an from January 2013 to December 2014 by retrospective analysis.

METHODA total of 224 specimens were collected from clinically diagnosed HFMD cases who were characterized by widespread mucocutaneous bullous reactions in Xi'an Children's Hospital from January 2013 to December 2014, the identification and subtyping of the isolates were conducted with real-time fluorescent quantitative RT-PCR. A retrospective analysis was performed to analyze the clinical presentation, laboratory tests and late follow-up problems of the HFMD.

RESULTIn the clinically diagnosed HFMD cases who were characterized by widespread mucocutaneous bullous reactions, 207 were caused by coxsackievirus A6 (CA6), accounting for 92. 4% of all cases with bullous, 4 were caused by enterovirus 71 (EV71), accounting for 1.8%, 10 were caused by coxsackievirus A16 (CA16), accounting for 4. 5%; 4 cases were negative for these viruses. In the cases positive for intestinal virus-nucleic acid, 130 were male, 90 were female; male to female ratio was 1. 44: 1, 203 were <5 years old, accounting for 92. 3%. Leukocytosis was found in 75 cases (34. 1%); high-sensitivity C-reactive protein (hsCRP) increased in 200 cases (90. 9%); elevated myocardial enzyme CK-MB was found in 35 cases (15. 9%), alanine aminotransferase increased in 15 cases (6. 8%); 187 cases had fever (85. 0%). None of the cases had serious complications such as encephalitis or myocarditis. In the course of the critical phase bullous rash or large vesicle-like changes, obvious itching, and facial rash appeared. After the fluid in the bullae was absorbed or the bullae ruptured or became ulcerated, scar formation and large areas of exfoliation occurred, with no effusion on the newly formed epithelium in the base, without significant pigmentation on later follow-up. In the late follow up process, 52 cases in CA6-positive patients (25. 1%) developed onychomadesis within 2-4 weeks after onset, 1 to 8 nails, an average of 4. 3 fell off, new nails grew, the nail bed showed no structural abnormalities and hyperplasia after falling off, the surface was smooth, had no hypertrophy, left no sequelae.

CONCLUSIONThe pathogen in HFMD characterized by widespread bullous reactions was mainly the CA6, this kind of HFMD was mainly mild type, with significant itching, later the bullae may have scar formation and skin exfoliation, in some cases onychomadesis may occur.

Child ; Enterovirus A, Human ; Enterovirus Infections ; pathology ; Exanthema ; pathology ; Female ; Fever ; Hand, Foot and Mouth Disease ; pathology ; Humans ; Male ; Pruritus ; Retrospective Studies

AIM:To investigate the effects of thalidomide ( THD) on the activation of connective tissue growth factor ( CTGF) gene promoter induced by transforming growth factor β1 ( TGF-β1 ) in human embryonic lung fibroblasts ( HELF) .METHODS:DNA sequence of CTGF gene promoter was cloned into luciferase reporter gene vector to construct the recombinant eukaryotic expression vector pGL 3-CTGFP, and the recombinant vector was transfected into HELF cell line.The effects of TGF-β1 and THD on the activation of CTGF gene promoter were detected by dual-luciferase analysis . RESULTS:TGF-β1 increased the reporter gene activity dose-dependently (P<0.05), with a plateau at 5 μg/L being 2.16 folds as high as the control .TGF-β1-induced increase in the reporter gene activity was also time-dependent ( P<0.05).After exposure to TGF-β1(5 μg/L), the level of luciferase activity reached its peak at 12 h and was 2.52 folds as high as the control .THD significantly inhibited TGF-β1-induced increase in the reporter gene activity in a dose-dependent manner , but its basal activity was not changed .CONCLUSION: TGF-β1 stimulates the transcriptional activity of CTGF gene promoter in HELF cells in a dose-and time-dependent manner , while THD may inhibit the effects dose-dependently .

AIM:To observe protective effect of L-glutamine on myocardial cell injury induced by endotoxin. METHODS:22 male SD rats,weighting(250?30)g,were randomly divided into three groups:control group( n= 7),endotoxin group( n= 7),glutamine/endotoxin group( n= 8). Heart were isolated from rats and perfused on Langendorff apparatus with Krebs- Henseleit(K-H)buffer(Saturation 95%O 2+5%CO 2)at a constant pressure(8.33 kPa)and temperature(37℃). The activity of superoxide dismutase (SOD)and malondialdehyde (MDA)content of efflux from coronary artery, monophasic action potential(MAP)and contractile force were measured at certain timepoints (0 min,20 min,50 min,80 min). RESULTS: Monophasic action potential and contractile force were markedly improved in Gln/ET group. The activity of SOD and MDA level in Gln/ET group restored closely to that in control group. CONCLUSION:L-gluta mine protects myocardial cells from injure induced by endotoxin.

AIM: To investigate the protective effect of Siduqing, a Chinese medicine, on LPS-induced myocardium injury in mice and its mechanisms. METHODS: Mice were divided into 4 group: control, LPS, Siduqing treatment and Siduqing group, and administered intragastrically with Siduqing decoction or distilled water (0 2 mL/10 g) twice a day for 3 days, two hours after Chinese herbal medicine treatment on day 3, LPS (30 mg/kg) or normal saline was injected intraperitoneally. The serum creatine kinase (CK) and myocardial superoxide dismutase (SOD) activities were determined, and myocardial tumor necrosis factor ? (TNF?) and malondialdehyde (MDA) contents were also detected. In addition, the histological changes and ultrastructure of heart were examined. RESULTS: Histological examination showed edema in myocardium and architectural disarray at 12, 24 h after LPS injection, mitochondrial swelling, condensation and margination of chromatin, irregular nuclear envelope and loss of contractile filaments at 24 h post LPS administration, while Siduqing treatment attenuated the above pathological changes of myocardium. CK activity in serum and myocardial TNF? content were higher in LPS group than control and Siduqing treatment group. Myocardial SOD activity in siduqing treatment group was higher than that in LPS group, but there was no difference in myocardial MDA content between control, LPS and Siduqing treatment group. CONCLUSION: These data suggest that Siduqing protects myocardium against LPS- induced injury via inhibiting myocardial TNF? production.