Acute Disease ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Muscle Weakness ; chemically induced ; epidemiology ; Organophosphate Poisoning ; Pesticides ; poisoning ; Respiratory Insufficiency ; chemically induced ; epidemiology ; Risk Factors

Acute Disease ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Organophosphate Poisoning ; Pesticides ; poisoning ; Pneumonia, Ventilator-Associated ; etiology ; Retrospective Studies ; Risk Factors ; Young Adult

Objective To establish the rapid purification of Coxsackievirus A16 using ultracentrifugation .And To prepare and i‐dentify the neutralizing monoclonal antibody against CA16 .Methods The CA16 culture supernatant was harvested and then con‐centrated by 100K capsule .The concentration of CA16 was purified by cesium chloride ultracentrifugation .Purification of CA16 were identified by transmission electron microscopy .BALB/c mice were immunized with inactivated CA16 .Spleen cells were harves‐ted and fused with SP2/0 myeloma cells ,hybridoma cell strain secreting mAb against CA16 were objected to screening .Character‐ization of the prepared mAb were analyzed by ELISA and microneutralization assay .Results The purified CA16 method of cesium chloride gradient ultracentrifugation was established ,TEM analysis was showed that CA16 particles have icosahedral structure ,the diameters of the viral particles were approximately 20-30 nm .Two hybridoma cell strains secreting mAb against CA16 were ob‐tained ,the subtypes of two mAbs were IgG2a ,the binding titers of Anti/CA16/5 and Anti/CA16/10 were 103 and 104 respectively . Neutralizing titer of the two mAbs were 1∶256 and 1∶1 024 respectively .Conclusion Establishment method of cesium chloride gradient ultracentrifugation was performed to purify CA16 ,the two mAbs with neutralizing ability to against CA16 may become ap‐plication of treatment and vaccine .

To study the kinetics in vivo of a Hantaan virus DNA vaccine, we constructed a fusion DNA vaccine,pEGFP/S, by cloning the S segment of Hantavirus into the vector, pEGFP-C1, which encodes Green fluorescent protein EGFP. In this report, we provide evidence that pEGFP/S was distributed and persistently expressed for more than 60 days in several organs after inoculation. Our findings suggest that the persistent immune responses induced by a Hantaan virus DNA vaccine are likely due to the plasmid pEGFP/S deposited in vivo, which acts as a booster immunization.

The surface-enhanced Raman spectra of bilirubin and biliverdin were obtained.By the bands analysis of the spectra,the orientation of bilirubin and biliverdin on the surface of silver colloid was discussed.In such case,the bilirubin was adsorbed on the silver colloid particle with the two planar pyrromethenone groups intercalated into the globe silver colloid particle,however,the biliverdin might lie flat on the surface of silver colloid with syn-synsyn conformation.

Objective To investigate the abnormal expression of microRNA (miRNA)-25 in the serum of gastric cancer patients and its clinical significance. Methods In Xijing Hospital of Digestive Diseases,Fourth Military Medical University,86 gastric cancer patients with operation and completed follow-up data,70 gastric adenoma patients and 80 healthy controls were selected as study objects.Total RNA was isolated from the serum. After the stable and sensitive miRNA-25 absolute quantity detection method established,the serum levels of miRNA-25 in gastric carcinoma patients,gastric adenoma patients and healthy controls were tested according to this method. The expression differences of miRNA-25 in the serum of patients with gastric cancer and gastric adenoma and healthy controls were analyzed with statistic analysis,and the correlation between miRNA-25 expression level and clinic pathological features of gastric cancer was also analyzed. Results The expression level of miRNA-25 in the serum of gastric cancer patients (135. 6 fmol/μg total RNA) was significantly higher than that of gastric adenoma patients (67. 7 fmol/μg total RNA) and healthy controls (62. 2 fmol/μg total RNA)(P＜0. 01). The receiver operating characterisstic curve of miRNA-25 indicated that serum miRNA-25 with good specificity and sensitivity in gastric cancer diagnosis (AUC=0. 827). The serum level of miRNA-25 in gastric cancer patients with lymph node metastasis [(148. 3±10. 2) fmol/μg total RNA] or clinicopathological stage Ⅲ /Ⅳ patients [(146. 7±9.5) fmol/μg total RNA] was significantly higher than that of gastric cancer patients without lymph node metastasis [(120. 3±10. 1)fmol/μg total RNA] or clinicopathological stage Ⅰ/Ⅱpatients [(119. 4±12. 2) fmol/μg total RNA] (P＜0.05). The correlation statistical analysis result indicated that there was no significant difference in survival period between serum miRNA-25 highly expressed and lowly expressed gastric cancer patients (P＞0. 05).Conclusion Serum miRNA-25 testing maybe helpful in diagnosis and prognosis of gastric cancer.

Objective To investigate the effect of integrin-?1 antisense oligodeoxynucleotide(ASODN) on(human) pancreatic cancinoma transplanted subcutaneously in nude mice.Methods The models of human(pancreatic) cancinoma transplanted subcutaneously were established in nude mice,then divided randomly into 3 groups and different treatment was given respectively(control group,random oligodeoxynucleotide group and ASODN group).After treatment,the weight of nude mice and tumor volume were observed,and the tumor growth inhibitory rate and the tumor response rate were calculated.The expressions of integrin-?1 mRNA and protein in tumor tissue were determined by RT-PCR and Western-blot.Results The tumor growth inhibitory rate in the random oligodexynucleotide group and the ASODN group was 4.75% and 72.70%,respectively.The tumor decrease rate of the ASODN group was 10.91%.The expression level of integrin-?1 mRNA and protein was decreased in the ASODN group compared with other 2 control groups. Conclusions Our findings suggest that integrin-?1 antisense oligodeoxynucleotides result in marked inhibition of human pancreatic(cancinoma) growth in nude mice.It may be a novel treatment approach for human pancreatic carcinoma.

OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

OBJECTIVETo study the effect of Zhibai Dihuang Pill (ZBDHP) on urokinase-type plasminogen activator (uPA) and sperm quality in ureaplasma urealyticum (Uu) infection infertile patients.

METHODSRecruited were 80 infertility patients with Uu infection at Andriatrics Clinics and Department of Reproduction, including 130 cases of positive Uu semen and 50 cases of negative Uu semen. Patients with positive Uu semen were randomly assigned to the observation group (72 cases) and the control group (58 cases) according to the visit sequence. All patients took antibiotics for 2 weeks. Patients in the observation group additionally took ZBDHP, 6 g each time, twice daily. Those in the control group additionally took Vit E (100 mg each time, twice per day) and ATP (40 mg each time, twice per day). The therapeutic course for all was 90 days. Semen parameters and uPA contents of the sperm membrane were detected and comparatively analyzed.

RESULTSThe sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm in Uu positive infected patients were lower than those in Uu negative infected patients with statistical difference (P < 0.05), but with no significant difference in the sperm density between the two groups (P > 0.05). There was no statistical difference in pre-treatment sperm membrane uPA contents and sperm parameters between the two groups (P > 0.05). Compared with before treatment in the same group, the sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm obviously increased in the two groups with statistical difference (P < 0.05). After treatment, the sperm membrane uPA content increased more obviously in the observation group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSInfection with Uu leads to decreased uPA content of sperm membrance and the sperm motility. ZBDHP could effectively treat Uu infected infertility possibly through fighting against Uu damaged sperm membrane and make the sperm membrane uPA content return to normal, and elevate the fertilizability of sperms.

Anti-Bacterial Agents ; administration & dosage ; pharmacology ; therapeutic use ; Communicable Diseases ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Humans ; Infertility ; Infertility, Male ; Male ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum ; drug effects ; Urokinase-Type Plasminogen Activator ; metabolism

Objective Epithelial mesenchymal transition (EMT) has a role in the proliferation and metastasis of various types of cells.This study investigates the hepatic oval cell's mechanism of EMT induced by histone deacetylase inhibition and the resulting cell motility increase from EMT.Methods Hepatic oval cell stem cell markers and marker changes were detected by flow cytometry,and after histone deacetylase inhibition induced EMT,the morphological changes were recorded.Western blot and quantitative real-time polymerase chain reaction detected the expression of E-cadherin,vimentin and Snail.Furthermore,confocal microscopy analysis recognized the nuclear translocation of Snail.Results Flow cytometry revealed no changes in the stem cell properties of hepatic oval cells in the cell culture process.Oval cell EMT,induced by HDACi,was observed through morphological changes,a reduction of the epithelial cell marker E cadherin,and an increase of the mesenchymal cell marker Vimentin.HDACi can promote the expression and nuclear translocation of Snail,which is the key hepatic oval cell transcription factor for both EMT and enhanced motility.Therefore,Snail RNA interference can suppress HDACi induced EMT in hepatic oval cells.Conclusions In conclusion,histone deacetylase inhibition induces hepatic oval cell epithelial-mesenchymal transition by Snail.