Objective To evaluate the expression of KAII (CD82) gene inhibited by small interfering RNA (siRNA) in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B,C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAd1 lentivrus particle ( MOI =5) was also used to infect T3 cells. The expression of CD82 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ±0.242,1. 311±0.048, 0. 664 + 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CD82 mRNA in empty vector group and that of A, B, C, D groups was significant (P＜0.01 ). Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.

Objective To characterize the association between the mean CT value on a scout view and the dependent mA selection method, and to evaluate the clinical value of a mA selection method based on scout view mean CT value in obtaining individualized scan protocol and consistent image quality for patient population on 64-row MSCT CT coronary angiography (CTCA).Methods One hundred patients (group A) underwent CCTA consecutively using standard protocol with a fixed mA.The mean CT value of a fixed ROI (region of interest) from the scout AP view and the CTCA image noise (standard deviation on the root of ascending aorta) were measured.The correlation between CT values and noise was studied establish a formula and a list to determine the required mA for obtaining a consistent CTCA image noise based on the measured SV CT value.Another 100 patients (group B) were scanned using the same parameters as group A except the mA and the CT value was also measured.The mA was determined by the list established previously.The CTCA image quality (IQ) as well as the image noise (IN) and the effective dose (ED) from the two groups were statistically analyzed using t-test.The CT findings for the 32 patients in the group B were also compared with the selective coronary angiography (SCA) results.The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of CCTA for detection of significant stenosis were obtained.Results The formula between the required mA and the CT value was : XmA=FmA×[(K1×CTscout + C1)/INa]2.The CCTA images in B group had statistically higher IN (27.66±2.57,22.22±4.17,t=11.33, P=0.000), but no statistical difference between IQ scores for the two groups (3.29±0.66,3.37±0.67,t=0.009, P=0.990), and ED[(8.72±2.51) versus (12.53±0.90) mSv]was 30% lower for the B group (P<0.01).For the 32 patients in the B group who had SCA, the CCTA sensitivity, specificity, positive predictive value, negative predictive value, and stenosis detection accuracy were 94.92% (56/59), 92.13% (82/89) ,88.89% (56/63) ,96.47% (82/85) and 93.24% (138/148) wspectively for stenosis greater than 50%.Conclusion The mA selection method based on the SV CT value for CCTA in 64-row MSCT provides individualized protocol to obtain consistent image quality and to optimize dose delivery to patients.

Objective The study aimed to explore the relationship between AIF related pathway and the inju-ring of cultured SGNs (spiral ganglion neurons)by glutamate toxicity,and to find AIF expression and distribution changes in SGNs.Methods SGNs of 40 newborn rats within 3 day were obtained and cultured in vitro.Cultured cells were divided into four groups:the normal control group,10 mM,20 mM and 40 mM glutamate injured group, separately.After 48 h hours culturing,optical microscopy,immune fluorescence staining and real-time fluores-cence quantitative PCR were used to observe the morphology,AIF distribution,and AIF,calpain,Caspase3 expres-sion changes in SGNs in vitro.TUNEL was used to verify the cell apoptosis.ResuIts Noticeable morphological chan-ges and cell apoptosis were occurred in 20 mM glutamate group,with AIF nuclear translocation.AIF gene expression was significantly higher than normal after glutamate administration (P<0.05);moreover,calpain gene expression increased(P<0.05);but caspase3 expression was not statistically significantly increased in all glutamate treated groups (P>0.05). ConcIusion In the process of cultured SGNs injured by glutamate,AIF participated in the cell apoptosis.Noticeable cell apoptosis were occurred in 20 mM glutamate group with AIF nuclear translocation.Calpain up-expression also contributed to excitatory neurotransmitter injury on SGNs,but Caspase 3 had no obvious effects.

OBJECTIVE: This study was designed to find out the impact of micro-ecological environment on the incidence of allergic rhinitis after developing a model of allergic rhinitis on mice. METHOD: Sixty mice were randomly divided into GF group (n=30) and SPF group (n=30). Mice of GF group were fed in the germ-free environment and mice of SPF group were fed in the specific pathogen-free environment. Then each group were randomly divided into model group (20 mice) and control group (10 mice). Establish allergic rhinitis model in the mice of model group using ovalbumin (OVA) at the age of 6 weeks, observe and score the corresponding symptoms and signs that could been induced. Stain with hematoxylin eosin (HE) staining method for nasal mucosa to observe the morphological changes. Using enzyme linked immunosorbent assay to detect the concentration of IgE, IFN-γ and IL-4 in the peripheral blood serum. RESULT: The chi square test showed that the incidence of allergic rhinithis in the mice of GF group was significantly higher than that in the SPF group (P< 0. 05). HE staining showed that the nasal mucosas of allergic rhinitis positive reaction mice were highly congestive and edematous and had a large number of inflammatory cell infiltration, while there was no abnormal morphology of nasal mucosas in mice with no allergic rhinitis reaction. EOS counting displayed that the number of eosinophilic cells in nasal mucosa of positive allergic rhinitis reaction mice was increased significantly. The concentration of IgE and IL-4 in the serum of positive allergic rhinitis reaction mice was highly increased (P <0. 05), and IFN-γ was significantly decreased (P< 0.05). CONCLUSION The difference of micro-ecological environment may play a key role in the occurrence of allergic rhinitis in mice.
Animals ; Disease Models, Animal ; Environment ; Incidence ; Interleukin-4 ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; Ovalbumin ; Rhinitis ; Rhinitis, Allergic ; etiology

Objective To investigate the underlying mechnisms of Kangai-1(KAI1)gene,a tumor metastasis suppressor gene,on metastasis and prolification of pancreatic cancer cells.Methods The plasmin containing Ad-KAI1 was established and transfected into pancreatic cancer cell line PCNA Ⅰ.The PCNA Ⅰ cells were then divided into different groups according to the times induced by vascular endothelial growth factor(VEGF)and epidermal growth factor(EGF).The morphology and migrational ability of PANC Ⅰ cells were compared before and after transfection by microscopy and transwell method,respectively.The expressions of intercellular adhesion molecule-1(ICAM-1)and matrix metalloproteinases-9(MMP-9)in PANC Ⅰ cells were examined by immunocytochemistry before and after transfection.Results The migrational ability of PCNA Ⅰ cells transfected with Ad-KAI1 was siginificantly decreased compared with untransfected PCNA Ⅰ cells(P＜0.05).Immunocytochemistry study revealed that the expressions of ICAM-1 and MMP-9 were both positive in untransfected PCNA Ⅰcells,but were both negative in transfected PCNA Ⅰ cells.Conclusion The inhibitory mechanism of KAI1 gene on metastasis of pancreatic cancer is associated with down-regulation of ICAM-1 and MMP-9expressions.

OBJECTIVE: To study the value of the pepsin in the sputum for diagnosing and evaluating the effectiveness of treatment of laryngopharyngeal reflux. METHOD: Thirty-six patients with the symptoms of dry pharynx, globus pharyngeus, excessive throat clearing, chronic cough and so on were divided into laryngopharyngeal reflux group and chronic laryngitis group by the results of therapeutic trial taking proton pump inhibitors for 3 months. The estimation of the reflux symptom index (RSI), the reflux finding score (RFS) and the detection of pepsin in the sputum were done before and after the treatment. The difference between two groups and the value of the pepsin were analyzed. RESULT: There were significant decreasing in RSI, RFS and pepsin level (P < 0.01) after the treatment in all patients. There were statistical differences between the laryngopharyngeal reflux group and the chronic laryngitis group in the changes of RSI and pepsin level (P < 0.01). CONCLUSION Pepsin level in the sputum might be used as a objective, effective method for diagnosing and evaluating the effectiveness in laryngopharyngeal reflux.
Adult ; Female ; Humans ; Laryngopharyngeal Reflux ; diagnosis ; therapy ; Male ; Middle Aged ; Pepsin A ; analysis ; Sputum ; chemistry ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the effect of thymic stromal lymphopoietin (TSLP) on transformation of dendritic cell (DC) and T cell in vitro.

METHODSMouse-derived immature dendritic cells and T lymphocytes were co-cultured in vitro, which were divided into 4 groups (TSLP stimulation group, TSLP stimulation and its receptor blocking group, ovalbumin stimulation group and ovalbumin stimulation and TSLP receptor blocking group). IL-4, IL-8 and IFN-β in cell culture supernatant were detected after 2 days by ELISA. SPSS 13.0 software was used to analyze the data.

RESULTSIL-4 levels of TSLP receptor blocking groups [(48.84 ± 1.56) pg/ml, (52.53 ± 2.36) pg/ml]were significantly lower than those of corresponding TSLP stimulation group and ovalbumin stimulation group [(72.55 ± 7.76) pg/ml, (80.47 ± 21.93) pg/ml;t = 5.994, P < 0.05;t = 2.534, P < 0.05]. However, there were not significant differences of IL-8 and IFN-β expression between corresponding two groups of whether or not TSLP receptor blocking (all P > 0.05).

CONCLUSIONTSLP receptor blockade in vitro can inhibit T lymphocyte transformation to Th2, which may provide a new therapeutic strategy for clinical Th2 dominant diseases such as allergic rhinitis and asthma.

Animals ; Cell Transdifferentiation ; Cells, Cultured ; Coculture Techniques ; Cytokines ; antagonists & inhibitors ; Dendritic Cells ; drug effects ; Mice ; Mice, Inbred BALB C ; Th2 Cells ; cytology

Objective:To investigate and analyze disease status and risk factors of venous thromboembolism (VTE) during pregnancy and puerperium in our country.Methods:Clinical datas were collected from 575 patients diagnosed with VTE during pregnancy and puerperium and hospitalized in nine medical institutions in our country from January 1, 2015 to November 30, 2019, and retrospectively analyzed it′s disease status and risk factors.Results:(1) The proportion of VTE in pregnancy and puerperium was 50.6% (291/575) and 49.4% (284/575), respectively. Four patients died, the mortality rate was 0.7% (4/575). The cause of death was pulmonary embolism. (2) The location of VTE during pregnancy and puerperium was mainly in the lower limb vascular (76.2%, 438/575), followed by pulmonary vessels (7.1%, 41/575). (3) In the risk factors of VTE, cesarean section accounted for 32.3% (186/575), maternal advance age accounted for 27.7% (159/575), braking or hospitalization during pregnancy accounted for 13.6% (78/575), other risk factors accounted for more than 5% were previous VTE, obesity, preterm birth, assistant reproductive technology conception and so on, pre-eclampsia and multiple pregnancy accounted for 4.9% (28/575) respectively. In addition, some patients with VTE did not have any of the above risk factors, and the incidence rate was as high as 23.1% (133/575).Conclusions:The occurrence of VTE during pregnancy and puerperium is related to multiple risk factors, and could lead to matemal death, It is very necessary to screen VTE risk factors for all pregnant women, to make corresponding prevention and control measures.

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.

ObjectiveTo describe the epidemic characteristics of COVID-19 after policy adjustment from “Category B notifiable disease with category A management” to “Category B notifiable disease with category B management”， and to explore the protective effect of previous infection with SARS-CoV-2 on common symptoms of reinfection. MethodsHealthcare workers infected with SARS-CoV-2 in a grade A tertiary hospital in Shanghai were included in the study from December 4， 2022 to January 11， 2023. Data on demographic characteristics， clinical symptoms， medical history， and COVID-19 vaccination history were collected. We determined the epidemiological curve and characteristics， and then compared the difference in the severity of clinical symptoms between primary and reinfection subjects. ResultsA total of 2 704 cases were included in the study， of which 45 had reinfection， 605 （22.4%）were males， 608 （22.5%）were doctors， 1 275 （47.2%） were nurses， and 2 351 （86.9%） received ≥3 doses of COVID-19 vaccination. The average age of these healthcare workers was （34.9±9.1） years old. The number of cases with mild/moderate illness， asymptomatic infection， fever， headache， dry cough， expectoration， and chest tightness were 2 704 （100.0%）， 92 （3.4%）， 2 385 （88.2%）， 2 066 （76.4%）， 1 642 （60.7%）， 1 807 （66.8%）， and 439 （16.2%）， respectively. Reinfection was a protective factor for fever （OR=0.161， P<0.001）， headache （OR=0.320， P<0.001）， and peak body temperature （β=-0.446， P<0.001）. ConclusionFollowing the COVID-19 policy adjustment as a category B notifiable disease， healthcare workers at a grade A tertiary hospital in Shanghai predominantly experiences mild to moderate COVID-19 symptoms. Reinfection results in milder clinical manifestations， with a lower proportion of being asymptomatic.