Objective:To study the effect of astrocyte elevated gene-1 (AEG-1) on proliferation and metastasis of papillary thyroid cancer (PTC) by regulating cell autophagy and epithelial-mesenchymal transition (EMT).Methods:Normal thyroid cells Nthy-ori3-1 and PTC cells TPC-1, FTC-133, B-CPAP and SW579 were cultured in vitro. Real time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of AEG-1 in PTC cells. The PTC cells with the highest AEG-1 expression were selected for AEG-1 shRNA infection, and then divided into sh-NC group and sh-AEG-1 group. Cell counting kit-8 (CCK8) and EdU (5-ethynyl-2 ′- deoxyuridine) experiments were used to detect the effect of AEG-1 on the proliferation of PTC cells; Transwell test was used to detect the effect of AEG-1 on the metastasis of PTC cells; subcutaneous tumorigenesis test in nude mice was used to detect the effect of AEG-1 on the expression of autophagy related proteins and EMT related proteins. PTC cells in sh-NC group and sh-AEG-1 group were treated with rapamycin and transforming growth factor-β (TGF-β), respectively. CCK8 and transwell assay were used to detect the cell proliferation and metastasis ability of each group of cells, respectively. Results:Compared with normal thyroid cells Nthy-ori3-1, the expression level of AEG-1 in PTC cells was increased, with the highest expression in TPC-1 cells. After AEG-1 shRNA was transfected into TPC-1 cells, cell proliferation, metastasis and tumorigenicity in vivo were reduced ( P<0.05). Compared with the sh-NC group, the expression of autophagy-related proteins P62 and Beclin1 were increased and the expression of LC3B protein was decreased, and EMT-related proteins E-cadherin expression were increased and N-cadherin and Vimentin protein expression were decreased in the sh-AEG-1 group ( P<0.05). CCK8 and Transwell experiments showed that treatment with autophagy inducer Rapamycin and EMT inducer TGF-β attenuated the inhibitory effect of sh-AEG-1 on proliferation and metastasis ability of PTC cell ( P<0.05). Conclusions:AEG-1 promotes the proliferation and metastasis of PTC cells by inducing cell autophagy and EMT.

Objective To observe the levels of N-terminal pro-brain natriuretic peptide (NT-proBNP),high density lipoprotein cholesterol (HDL-C) in predicting cardiovascular damage in Kawasaki disease (KD).Methods Enzyme-linked fluorescence analysis (ELFA) technique was used to measure serum NT-proBNP levels in 406 KD patients [including 104 cases of incomplete Kawasaki disease (IKD)] at the acute phase,the convalescent stage,at the same time,the blood HDL-C,Albumin,globulin,alanine aminotransferase (ALT),C-reactive protein (CRP),red blood cell sedimentation rate (ESR),blood white cell count (WBC),hemoglobin (Hb),blood platelet count (PLT) level were tested.According to the results of echocardiography,electrocardiogram,myocardial enzymes in KD,patients were divided into two groups:two hundred and twenty-two with cardiovascular damage and 184 without cardiovascular damage group.The age,gender,fever,the first application of the intravenous gamma globulin,laboratory markers of clinical features observed above the detection levels were compared,and these parameters for each group were compared using t test or analysis of variance,the single factor which was statistically significant were received operating characteristic curve (ROC) analysis.Then the cardiovascular damage group was divided into echocardiography abnormal group and echocardiography normal group,and t test was adopted to compare the clinical parameters of the two groups.Results ① The age,lip and oral changes,the first application of intravenous gamma globulin,blood NT-proBNP,HDL-C,albumn and ALT was significantly different between the cardiovascular damage group and non cardiovascular damage group (t/x22=4.989,4.721,6.212,18.834,12.788,4.851,4.541,All P＜0.05).② When the blood NT-pro-BNP was higher than 786.5 ng/L,its sensitivity and specificity for differentiating KD with cardiovascular damage was 86.5％ and 84.8％,respectively.When the blood HDL-C was lower than 0.655 mmol/L,its sensitivity and specificity for differentiating KD with cardiovascular damage was 80.4％ and 69.4％ respectively.When the blood NT-proBNP was higher than 786.5 ng/L in addition to HDL-C lower than 0.655 mmol/L,the specificity for differentiating KD with cardiovascular damage was 91.8％.③ For the 222 cases with cardiovascular dam-age,their blood NT-proBNP,HDL-C levels were statistically significantly different between the echocardiogra-phy abnormal group and echocardiography normal group (t=3.354,4.084,All P＜0.05).④ The serum NT-proBNP,ALT levels of the 406 acute and convalescent KD patients were significantly higher than the recovery phase.The blood HDL-C,albumin level of acute patients were significantly lower than those at the recovery phase,the difference was statistically significant (t=22.335,4.951,20.334,15.073,All P＜0.05).⑤ One hundred and four children with IKD were divided into patients with cardiovascular damage (74 cases) and without cardiovascu-lar damage (30 cases),the age,lip and oral changes,the first application of intravenous gamma globulin,blood NT-proBNP,HDL-C,albumin and ALT were significantly differentbetween these two groups (t=3.083,2.157,6.423,6.409,3.649,8.658,All P＜0.05).Conclusion Blood NT-proBNP and HDL-C are good pre-dictive parameters in children with cardiovascular damage of KD,IKD.

Objective To study the mechanism of Liangruntongluo Recipe in improving gastrointestinal function on DM rats induced by STZ. Methods Forty male Wistar rats were divided into four groups randomly: group A as model control group,group B as Liangruntongluo group,group C as Cisapride group,and group D as normal control group. There are 10 rats in each group. After the establishment of DM animal model with Wistar rats,the animals were raised for 6 weeks without any hypoglycemic agents. And then medicines were intragastric administrationed for another 6 weeks. In the 12th week motile and cholecystokinin in blood and tissue were tested. Result By the time of the twelfth week,the rats with DM presented the disorder of gastrointestinal hormone. Liangruntongluo Recipe affected the excretion and releasing of hormone in blood,stomach,and duodenum. With a wide target range,effect of Liangruntongluo Recipe is superior to Cisapride. Conclusions Chinese herbal medicine Liangruntongluo recipe has the superiority in improving the obstacle of the gastrointestinal function of diabetes mellitus.

Objective To analyze the clinical features of patients with acute stage brucellosis in Shenzhen,and provide a scientific basis for prevention and control of brucellosis in immigrant city.Methods A retrospective analysis was conducted to collect clinical data of patients with brucellosis admitted to the Department of Infectious Diseases,Shenzhen People's Hospital from May 2013 to May 2018.The patient's epidemiology manifestations,pathogen and laboratory examination results,diagnosis and treatment outcomes and prognosis were analyzed.Results Among the 39 patients with brucellosis,males were predominant,with a male to female ratio of 1.4 ∶ 1.0 (23 ∶ 16),an age of (44.91 ± 17.18) years and 24 cases were non-Guangdong natives.There were 23 cases with epidemiological history,including 14 cases with mutton,sheep viscera and goat milk history;the disease occurred throughout the year,mainly from February to July,a total of 26 cases.The clinical manifestations of the patients were mainly fever,sweating,fatigue,joint and muscle pain,weight loss,and liver or spleen or lymph nodes swelling.The blood culture was identified as 38 cases of Brucella melitensis and 1 case of Brucella suis.All strains were sensitive to common antibiotics in vitro.All cases were diagnosed as acute stage of brucellosis,2 cases with orchitis,1 case with brucellosis meningoencephalitis,3 cases with spondylitis,and 3 cases with misdiagnosis.Thirty-nine patients were cured according to the "Brucellosis Diagnosis and Treatment Guidelines (Trial)" and were followed up for 1 year.Conclusions Patients with brucellosis in Shenzhen are mainly infected with Brucella melitensis;fever,sweating,joint and muscle pain are the main clinical symptoms;the patient's efficacy and prognosis are better after treatment;for the occurrence of occasional misdiagnosis,it is recommended that in immigrant cities,medical staff should strengthen their understanding of brucellosis.

Objective:To discuss the effect of human umbilical cord mesenchymal stem cells culture supernatant(hUCMSCs-Sup)on the proliferation,apoptosis,and endometrium receptivity of the human endometrial stromal cells(hEndoSCs)treated with mifepristone(Ms),and to clarify the possible mechanism.Methods:The hEndoSCs were cultured in vitro and divided into control group and 40,60,80,and 100 μmol·L-1 Ms groups.The survival rates of the cells in various groups were detected by MTT assay.The hEndoSCs were divided into control group,40 μmol·L-1 Ms group,and 60 μmol·L-1 Ms group.The apoptotic rates of the cells in various groups were detected by flow cytometry;the expression levels of apoptosis-related protein B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)proteins in the cells in various groups were detected by Western blotting method,and the ratio of Bcl-2/Bax was calculated.After treated with hUCMSCs-Sup,the hEndoSCs were divided into control group,Ms group,Ms+hUCMSCs-Sup group,and Ms+hUCMSCs-Sup+3-methyladenine(3-MA)group.The survival rates of the cells in various groups were detected by MTT assay;the apoptotic rates of the cells in various groups were detected by flow cytometry;the expression levels of microtubule-associated protein 1 light chain 3B-Ⅱ(LC3B-Ⅱ)and microtubule-associated protein 1 light chain 3B-I(LC3B-Ⅰ)proteins in the cells in various groups were detected by Western blotting method,and the ratio of LC3B-Ⅱ/LC3B-Ⅰwas calculated;the expression levels of endometrium receptivity marker molecules mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.Results:Compared with control group,the survival rates of the cells in 40,60,80,and 100 μmol·L-1 Ms groups were significantly decreased(P<0.05)in a time-dependent and dose-dependent manner.Compared with control group,the apoptotic rates of the cells in 40 and 60 μmol·L-1 Ms groups were significantly increased(P<0.05),and the ratios of Bcl-2/Bax were significantly decreased(P<0.05).After treated with hUCMSCs-Sup,compared with control group,the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms group were significantly decreased(P<0.05),the apoptotic rate was significantly increased(P<0.05),and the expression levels of homeobox A10(HOXA10),leukemia inhibitory factor(LIF),and integrin subunit beta 3(ITGB3)mRNA in the cells were significantly decreased(P<0.05);compared with Ms group,the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰin the cells in Ms+hUCMSCs-Sup group were significantly increased(P<0.05),the apoptotic rate was significantly decreased(P<0.05),and the expression levels of HOXA10,LIF,and ITGB3 mRNA in the cells were significantly increased(P<0.05);compared with Ms+hUCMSCs-Sup group,the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms+hUCMSCs-Sup+3-MA group were significantly decreased(P<0.05).Conclusion:hUCMSCs-Sup can increase the survival rate and decrease the apoptotic rate of the hEndoSCs after treated with Ms,and increase the endometrium receptivity,and its mechanism may be associated with the activation of autophagy of the hEndoSCs by hUCMSCs-Sup.

OBJECTIVE To establish a method for simultaneous determination of 15 bile acids in Tongren niuhuang qingxin pills， and to determine the contents of 15 batches of samples. METHODS Using dehydrocholic acid as internal standard， the determination was performed by ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method. The determination was performed on Hypersil GOLD C18 column with methanol-0.1% formic acid solution as the mobile phase by gradient elution at the flow rate of 0.2 mL/min. The column temperature was 40 ℃ ， and the sample size was 2 µL. Using heated electrospray ion source， parallel reaction monitoring mode scanning was performed in negative ion mode. SPSS 24.0 software was used for chemical pattern recognition analysis of content determination results. RESULTS The 15 bile acid components had a good linear relationship with peak area （all R2≥0.998 9）； their precision， repeatability and stability were all good （all RSD≤5.49%）； the average recoveries were 93.8%-105.7% （RSD was 0.5%-5.8%）. The average contents of taurocholic acid， 7-oxodeoxycholic acid， 12-dehydrocholic acid， glycocholic acid， 3-oxo-7α， 12α-hydroxy-5β-cholanoic acid， taurochenodeoxycholic acid， 3α-hydroxy- 7-oxo-5β -cholanic acid， hyocholic acid， taurodeoxycholic acid sodium salt hydrate， hyodeoxycholic acid， cholic acid， glycochenodeoxycholic acid， glycodeoxycholic acid， chenodeoxycholic acid and deoxycholic acid were 670.56， 25.97， 10.54， 280.12， 4.04， 29.81， 182.98， 813.55， 120.95， 220.31， 797.37， 18.37， 68.59， 30.13， 59.82 μg/g， respectively. Both cluster analysis and principal component analysis divided 15 batches of Tongren niuhuang qingxin pills into 2 categories， S1-S12 as one category and S13-S15 as the other category. CONCLUSIONS The established method is accurate， sensitive and specific， and can determine many types of bile acids. It also can quickly achieve the quantitative analysis of 15 bile acids in Tongren niuhuang qingxin pills， which is suitable for the quality control of this drug.

Objective: To understand the epidemiological and clinical features of patients with hemorrhagic fever and renal syndrome (HFRS) in Shenzhen, and to accumulate experience in the diagnosis and treatment of HFRS in this area. Methods: A retrospective analysis was conducted by collecting the clinical data from 46 patients who were confirmed with HFRS and admitted to the Department of Infectious Diseases of Shenzhen People's Hospital from January 2015 to December 2018. The demographic characteristics, epidemiological, clinical manifestations, examinations, treatments and prognosis, and other characteristics were analyzed. Results: All the 46 patients with HFRS were residens in Shenzhen, with a male-to-female ratio of 6.67∶1.00(40∶6), aged (40.18 ± 15.63) years old, and 38 patients (82.61%) aged 23-45 years old. There were 41 patients (89.13%) with a history of HFRS epidemiology, and there were mice in their houses or workplaces. The houses of 39 patients (84.78%) were rented, and 34 patients(87.18%) rented their houses in urban villages. There were morbidity throughout the year, and 33 patients (71.74%) were ill from January to June. In clinical classification, 44 cases (95.65%) were mild, 2 cases (4.35%) were medium, and there were no severe or critical cases. The clinical manifestations were that all patients were hospitalized due to fever mainly with hyperthermia. Thirty-nine patients (84.78%) were presented with systemic aches, headaches, low back pain and eyelid pain, and 28 patients (60.87%) had skin and mucous membrane hyperemia flushing. Clinical stages showed that all patients had pyretogenesis stage and polyuria stage, including pyretogenesis stage [(7.34 ± 6.82) d], polyuria stage [(9.94 ± 5.77) d], only 4.35% (2/46) patients with hypotension shock stage, all patients did not have oliguric stage. On the next day of admission, the number of white blood cells in 46 patients was (8.17 ± 3.19) × 10⁹/L, and 38 cases (82.61%) in the normal range; platelet was (61.92 ± 32.53) × 10⁹/L, and 42 cases (91.30%) were decreased; the procalcitonin was (1.62 ± 0.38) ng/ml, and 41 cases (89.13%) were increased; C-reactive protein was (74.33 ± 30.48) mg/L, and 46 patients (100.00%) were elevated; creatinine was (176.25 ± 55.15) μmol/L, and 19 cases (41.30%) were increased. Abnormal liver function was manifested by increased enzymology, alanine aminotransferase was (137.58 ± 46.76) U/L, and aspartate aminotransferase was (129.82 ± 40.29) U/L. All patients were positive for Hantavirus IgM. B-ultrasound results showed that in 58.70% (27/46) patients, both kidneys were plump and the parenchymal echo was enhanced. Liver injury occurred in 39 patients (84.78%) and sinus bradycardia in 2 patients (4.35%). All the 46 patients were clinically cured. Conclusion The epidemiological characteristics of patients with HFRS in Shenzhen are typical, the clinical manifestations are mild, and the curative effect and prognosis are good.