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.

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

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 evaluate incidence of postoperative delirium after hip surgery in elderly patients by meta-analysis.

METHODSFrom January 1, 2014 to December 31, 2013, clinical literatures about postoperative delirium after hip surgery in elderly patients,were searched from the Pubmed. Literature extract table were formed according to inclusion and exclusion criteria. Stata-12.0 was applied for Meta-analysis. P was used to test heterogeneity of study, random-effect model was performed when I2 > 50%. Subgroup analysis was used according to stage of age, assessment scale of delirium and statistical area of literature. Begg test was used to test publication bias.

RESULTSTwenty-one literatures were included. Incidence of postoperative delirium after hip surgery in elderly patients by weighted and combination was 17% [95% CI (16%, 18%)]. Incidence of postoperative delirium after optional hip surgery was decreased more than emergency operation in included 5 literatures [OR = 0.32, 95% CI (0.22, 0.45)]. Incidence of postoperative delirium in patients less than 80 years old was 21% [95% CI (19%, 23%)], while 21% [95% CI (19%, 24%)] in patients more than 80 years old. Incidence of postoperative delirium in CAM evaluation scale was 23% [95% CI (21%, 26%)], while 19% [95% CI (17%, 21%)] in other evaluation scales. Incidence of postoperative delirium in Asian area was 17% [95% CI (15%, 20%)], while 23% [95% CI (21%, 25%)] in European and American area. There was no publication bias tested by Begg test (P < 0.05).

CONCLUSIONIncidence of postoperative delirium after hip surgery in elderly patients increases higher, especially in emergency operation. A standardizing research method is benefit for evaluate incidence of postoperative delirium after hip surgery in elderly patients, decreasing heterogeneity and publication bias.

Aged ; Delirium ; epidemiology ; Hip Fractures ; surgery ; Humans ; Incidence ; Postoperative Complications ; epidemiology ; Publication Bias

The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).