Objective To develop a chemiluminescence imaging DNA microarray method for simultaneous,quick and accurate detection of serotypes of human adenovirus (HAdV ),namely,HAdV3,HAdV7,HAdV11,HAdV14 and HAdV55.Methods Based on the specific gene sequences in the conserved region of adenovirus from GenBank, oligonucleotide primers and probes were designed and synthesized to prepare the oligonucleotide microarray.The specific genomic sequences were amplified by multiplex PCR method.The multiplex PCR amplification products were hybridized with the specific probes of microarray that was scanned after washing and chemiluminescenceb before the result was analyzed.After optimization of the multiple PCR system,hybridization reactions and conditions of chemiluminescence,the specificity,sensitivity and reproducibility of the chip were evaluated.Results The microarray displayed high sensitivity, specificity and reproducibility.The minimum detection limit of plasmidg DNA was 3 ×103 copies/reaction.The microarray detection results of 38 clinical samples were approximately consistent with those using the direct sequencing method(37 /38).Conclusion A chemiluminescence imaging DNA microarray method for quick,sensitive and specific detection of five serotypes of HAdV is established,which can provide a new means for detecting serotypes of HAdV.

Objective To report two cases of familial Gitelman syndrome and literature review regarding the updates of relevant genes,classification,treatment,and prognosis.Methods The clinical data of two sisters with Gitelman syndrome were retrospectively analyzed.Results Their blood pressures were within normal range.Hypokalaemic alkalosis,hypomagnesemia,and hypocalciuria were corrected almost completely after three days of intravenous magnesium and potassium infusion,spirolactone and indometacin.However,the maintenance of normal potassium was unsuccessful over one year.Conclusion Hypokalaemic alkalosis,hypomagnesemia,and hypocalciuria were normalized in Gitelman syndrome.There was some debate in regard to using PGE2 synthetase inhibitors.Tolerance of long-term medication will be the big challenge for curative effect.

Acupoint compatibility is a basic element for acupuncture prescription and an important link for clinical acupuncture workers to obtain a therapeutic effect. Studying factors influencing the effect of acupoint combination and ascertaining its concept and connotation are of important significance for studying the mechanism of acupuncture action. By sorting out, summing up and analyzing relevant modern literature, it is found that factors influencing the effect of acupoint combination are those affecting the therapeutic effects of combination of two or more than two acupoints. Its contents include five aspects: combination mode, combination effect, stimulation method, time factor and body status. Analysis and induction show that acupoint selection is the primary task in studying factors influencing the effect of acupoint combination.

Objective To develop a detection method based on the technology of gene chips which can quickly distinguish genes of Enterococcus faecalis, E.faecium and vancomycin resistance.Methods Based on the specific gene ( ddl) sequences of two types of Enterococcus from GenBank, oligonucleotide probes which could detect and distinguish special genes and drug resistance genes ( vanA,vanB) of Enterococcus were designed and compounded.Then,the probes were dotted to modified slide.The target DNA fragments of vancomycin-resistant Enterococcus ( VRE) were labeled with biotin by multiple PCR amplification, and then hybridized with oligonucleotide probes on slide.The results were analyzed by portable imager.The multiple PCR system, hybridization reaction and condition of the chemiluminescence method were optimized before the specificity, sensitivity and reproducibility of the chip were evaluated.Results One universal primer, four specific primers, one universal probe and four specific probes were selected.This gene chip was demonstrated of high specificity and repeatability.The detection sensitivity was 103 CFU/ml.The gene chip detection results of 10 clinical samples were basically consistent with the drug sensitivity test ( 8/10 ) .Conclusion A gene chip technique for the detection of VRE is established successfully.It is possible to distinguish the type of VRE and detect the genetic phenotypes of drug resistance by gene chip technique.

Objective To investigate the learning outcome of clinical scenarios discussion im-plemented in teaching medical students before clinical probation. Methods Using standardized pa-tients (SP) to simulate the clinical common difficult situation, all eight year program clinical medicine students (84) of Grade 2008 who were about to enter the clinical probation were given clinical commu-nication education guidance. Through five classification variables questionnaire and return visit, the teaching effect was evaluate. Epidata 3.0 was used to input data, and the SPSS 15.0 was used to make descriptive analysis of the questionnaire and proportion comparison. Results We found 35.4%(29) and 63.4%(52) of subjects liked and approval this training, respectively;90.2%(74) of the subjects thought the diffi-culty of this train was moderate; 77.3% (65) of subjects believed that it was necessary to set the clinical scenarios discussion before clinical probation. Conclusions The implementation of clinical scenarios discussion before clinical probation was effective on inducing the medical students to pay attention to the doctor-patient communication, and deep thinking about the communication notice mat-ters and cop-ing styles in clinical environment.

Primary insomnia (PI) is now a nonorganic sleep disorder that seriously affects the quality of life of people around the world and attracts the attention of medical circles at home and abroad. Western medicine treatment has side effects such as drug dependence and withdrawal rebound and cannot obviously improve the quality of sleep and life, while acupuncture has a good therapeutic effect on PI. This article explores the latest research status of acupuncture treatment for PI. Chinese Journal Net (CNKI, VIP and Wanfang) was mainly searched to retrieve and sum up recent five years’ literature about the study of acupuncture treatment for PI. The results show that acupuncture has an advantage in producing a short-term marked therapeutic effect on PI and being of a high total efficacy rate and can effectively treat primary insomnia.

ObjectiveTo investigate the condition of infants with urolithiasis caused by melaminetainted powdered formula at one-year follow-up. Methods Eighty-one young children with melamine-induced urolithiasis were followed up,which included urinalysis,renal-function tests,urinary tests for biochemical markers of renal glomerular and tubular function,and ultrasonography.Eighty-one age-matched healthy infants without exposure to melamine-contaminated formulas were enrolled as controls. Results Fifty-one of the patients were male and 30 female,with a mean age of 26 months (range 13 -48 months).The 81 children were divided into 2 groups according to treatment protocols.Group 1 received conservative treatment ( n =54 ) and Group 2 received surgical treatment ( n =27 ).In Group 1,3 patients (5.6％) had stone-residual:1 girl had calculus of 0.4 cm in diameter in the left kidney and 2 girls had calculus of 0.3 cm in diameter in the right kidney.In Group 2,2 patients (7.4％) had stone-residual,1 boy had crystallization of 0.2 cm in diameter in the right kidney and 1 boy had calculus of 1.5 cm in diameter in the left kidney,positive stone was confirmed by X-ray and accepted percutaneous nephrolithotomy.Urinary microalbumin levels were significantly increased in the children with melamine-induced urolithiasis compared with the control group (P ＜0.01 ).Blood urea nitrogen and serum creatinine were within the normal range with no significant difference between the 2 groups. ConclusionsAt one-year follow-up most of the children exposed to melamine-tainted infant formula in both treatment groups had returned to a healthy state,suggesting a satisfactory outcome for both treatment options.However,surgery is recommended in patients with stoneresidual ＞ 10 mm.

Objective To assess the clinical therapeutic effects and safety of Jiangtang Shuxin decoction (JTSXD) on diabetic patients complicated with chronic heart failure (CHF),and to search for its possible function mechanisms.Methods A prospective randomized controlled study was conducted,80 diabetic patients complicated with CHF [New York Heart Association (NYHA) functional class Ⅱ-Ⅲ] admitted into the Department of Traditional Chinese Medicine (TCM) or of Cardiology in Affiliated Hospital of Guangxi Youjiang National Medical College from October 2015 to September 2016 were enrolled,they were assigned to an observation group and a control group by randomized method with a computer,and finally 77 patients (39 cases in observation group and 38 cases in control group) completed this trial.The patients in control group received standardized routine western medical treatment,while the observation group was additionally administered JTSXD (including ingredients:astragalus 15 g,ginseng 10 g,radix ophiopogonis 15 g,radix rehmanniae 15 g,comus 10 g,rhizome coptidis 8 g,peach kernel 10 g,salvia mitiorrhiza 10 g,magnoliaceae 10 g,yam 15 g) on the basis of conventional therapy.The therapeutic course for all the patients in both groups was 2 months.Before and after treatment,the 6-minute walking distance (6MWD) was assessed;the TCM syndrome accumulated scores of the two groups were calculated;the left ventricular end-diastolic volume (LVEDV),the left ventricle ejection fraction (LVEF),the stroke volume (SV),the cardiac output (CO),and the maximum blood flow velocity of early diastolic/atrium late diastolic (E/A) were detected by echocardiography.The serum levels of glycosylated hemoglobin (HbA1c),angiotensin Ⅱ (Ang Ⅱ) and plasma B type brain natriuretic peptide (BNP) were tested with enzyme linked immunosorbent assay (ELISA);the level changes of total cholesterol (TC),triglyeride (TG),high density lipoprotein cholesteral (HDL-C) and low density lipoprotein cholesteral (LDL-C) were observed.Results Compared with the control group,after treatment in the observed group,the TCM syndrome score of palpitation,fatigue and thetotal accumulated score were all obviously decreased (palpitation score:0.9 ± 0.4 vs.1.2 ± 0.8,fatigue score:1.1 ± 0.7 vs.1.7 ± 0.8,total accumulated score:4.8 ± 1.2 vs.8.1 ± 1.8,all P ＜ 0.05);the LVEDV,the serum levels of HbA1c,Ang Ⅱ and BNP were also obviously decreased in the observed group [LVEDV (mL):136.28 ± 17.52 vs.158.82 ± 19.03,HbA1c (％):6.11±0.36 vs.6.89 ±0.32,Ang Ⅱ (ng/L):66.48 ± 17.64 vs.84.55 ± 20.39,BNP (μg/L):138.45 ± 87.55 vs.219.14±88.83,all P ＜ 0.05];The 6MWD,LVEF,SV,CO and E/A were all increased plainly in the observed group [6MWD (m):470.47 ± 79.66 vs.428.46 ± 88.56,LVEF:0.51 ±0.05 vs.0.46 ± 0.04,SV (mL):55.36 ± 2.88 vs.50.32±2.76,CO (L/min):5.74±0.91 vs.4.92±0.74,E/A:1.18±0.27 vs.0.83±0.28,all P ＜ 0.05].The degrees of decreased levels in TC,TG,LDL-C and the degrees of increased levels of HDL-C in observed group were superior to those of the control group,but there were no statistical significant differences (all P ＞ 0.05).Conclusion JTSXD shows good therapeutic effect and safety for treatment of diabetic patients accompanied by CHF (NYHA functional class Ⅱ-Ⅲ),and its mechanisms may be related to its regulation of glucose (reduction of HbA1c level),correction of lipid metabolism disorders,improvement of myocardial energy supply,inhibition of the activation of renin-angiotensin-aldosterone system (RAAS) and the secretion of BNP.

Objective To systematically review the clinical efficacy and safety of early use of recombinant human erythropoietin (rHu-EPO) for neuroprotection in premature infants.Method From establishment of the databases to November 2017,clinical randomized controlled trials (RCTs) of early use of rHu-EPO in premature infants were searched on English databases (PubMed,Embase,Cochrane Collaboration) and Chinese databases (CNKI,SinoMed,Wanfang,and VIP Database).Patients receiving conventional therapy were assigned into control group,and patients receiving both conventional therapy and rHu-EPO were assigned into rHu-EPO group.rHu-EPO group was subdivided into high-dose continuous group and low-dose intermittent group.We retrieved the related literatures,evaluated the quality,and then performed Meta-analysis using RevMan 5.3 software.Result A total of 2 588 patients in 17 studies were included and analyzed.Compared with the control group,Meta-analysis showed that early use of rHu-EPO was more effective in improving NBNA scores at 40 weeks of corrected gestational age (cGA) (MD=2.46,95％CI 1.58～3.33,P＜0.001),and high-dose continuous group was better than low-dose intermittent group (MD=3.19,95％CI 0.45～5.93,P=0.002;MD=2.22,95％CI 1.29～3.16,P＜0.001).Low-dose intermittent use of rHu-EPO significantly increased mental development index (MDI) (MD=6.66,95％CI 0.80～12.51,P=0.010) and psychomotor development index (PDI) (MD=5.77,95％CI 4.00～7.55,P＜0.001),and reduced the incidence of MDI＜70 at cGA 18～22 months (OR=0.42,95％CI 0.27～0.67,P＜0.001).As to the safety and side effects,treatment with rHu-EPO reduced the risk of necrotizing enterocolitis (OR=0.48,95％CI 0.31 ～0.72,P＜0.001) and periventricular leukomalacia (OR=0.52,95％CI 0.35～0.75,P＜0.001)without increasing the risk of bronchial dysplasia,patent ductus arteriosus,retinopathy in prematurity and intraventricular hemorrhage.Conclusion Current evidence shows that the early use of rHu-EPO in premature infants is relatively effective and safe,but multi-center RCTs are still needed.

Objective:To investigate the effect and possible molecular mechanism of mannan-binding lectin-associated serine protease (MASP) 1 on the biological behavior of hepatocellular carcinoma(HCC) cells.Methods:From January 10 to October 25, 2022, 20 pairs of fresh liver cancer tissue and adjacent (3 cm from cancer tissue) normal tissue samples of patients who underwent hepatic cancer resection at the Affiliated Hospital of Nantong University were collected, and the expression of MASP1 was analyzed. From March 1, 2012 to May 20, 2017, the cancer tissue and adjacent (3 cm from cancer tissue) normal tissue samples of 67 patients with HCC were collected from the tissue sample bank of the Department of Pathology, the Affiliated Hospital of Nantong University and the correlation between the expression level of MASP1 and clinical data of patients with HCC were analyzed. Human hepatoma cells lines SK-Hep1 and Hep3B were cultured, and MASP1 overexpression and MASP1 knockdown cell lines were constructed. SK-Hep1 negative control group, SK-Hep1 MASP1 overexpression group, Hep3B negative control group and Hep3B MASP1 knockdown group were set up. The effect of MASP1 on the proliferation of HCC cells was detected by subcutaneous tumor transplantation experiment in nude mice. The effect of MASP1 on the expression of epithelial-mesenchymal transition related proteins (N-cadherin, matrix metalloproteinase 9(MMP9), and E-cadherin), and the effects of MASP1 on the expression of phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway related proteins and their phosphorylation levels were detected by Western blotting. Independent sample- t test, paired- t test and chi-square test were used for statistical analysis. Results:The results of immunohistochemical staining of 20 pairs of fresh tissue samples showed that the expression level of MASP1 in liver cancer tissue was lower than that in adjacent normal tissue (3.73±1.03 vs. 6.76±1.46), and the difference was statistically significant ( t=16.97, P<0.001). The correlation analysis of MASP1 expression level and clinical data of 67 patients with HCC revealed that the expression level of MASP1 was related to vascular invasion of the tumor, and the difference was statistically significant( χ2=5.20, P=0.023). The subcutaneous tumor transplantation experiment in nude mice showed that the volume and weight of subcutaneous tumor of mice in SK-Hep1 MASP1 overexpression group were lower than those of the SK-Hep1 negative control group((165.42±149.08) mm 3vs. (260.42±179.78) mm 3, (0.13±0.12) g vs. (0.18±0.12) g), and the differences were statistically significant ( t=5.15 and 3.89, both P<0.05). The results of Western blotting showed that the expression levels of N-cadherin and MMP9 in SK-Hep1 MASP1 overexpression group were lower than those of SK-Hep1 negative control group, while the expression level of E-cadherin was higher than that of SK-Hep1 negative control group (0.73±0.01 vs. 1.02±0.02, 0.40±0.01 vs. 0.69±0.01, 0.91±0.02 vs. 0.78±0.02), and the differences were statistically significant ( t=24.23, 36.87 and 9.27, all P<0.001). The expression levels of N-cadherin and MMP9 in Hep3B MASP1 knockdown group were higher than those in Hep3B negative control group, and the expression levels of E-cadherin was lower than that in Hep3B negative control group (1.04±0.01 vs. 0.31±0.01, 0.54±0.02 vs. 0.04±0.01, 0.56±0.01 vs. 0.93±0.01), and the differences were statistically significant ( t=163.20, 53.16, 60.74, all P<0.001). The expression levels of phosphoinositide PI3K, phosphoinositide Akt, and phosphoinositide mTOR of SK-Hep1 MASP1 overexpression group were lower than those of SK-Hep1 negative control group (0.59±0.01 vs.1.02±0.01, 0.64±0.01 vs. 1.12±0.02, 0.10±0.01 vs. 1.05±0.02); the expression levels of phosphoinositide PI3K, phosphoinositide Akt, and phosphoinositide mTOR of Hep3B MASP1 knockdown group were higher than those of Hep3B negative control group (0.96±0.01 vs. 0.55±0.01, 0.99±0.01 vs. 0.38±0.01, 0.93±0.02 vs. 0.06±0.01), and the differences were statistically significant ( t=40.87, 40.91, 87.30, 44.17, 107.50, 67.28, all P<0.001). Conclusions:The expression level of MASP1 is low in HCC tissue, and is significantly correlated with the poor prognosis of HCC patients and the occurrence of tumor vascular invasion. MASP1 may affect the proliferation and migration of liver cancer cells by regulating the PI3K/Akt/mTOR signaling pathway, suggesting that MASP1 may become a key gene in the treatment of HCC.