Objective To construct a single chain Fv immunotoxin HAb25(scFv)-PE40 and explore its targeting effects on human hepatocellular carcinoma(HCC) cells.Methods First,the HAb25(scFv) gene was ligated with PE40,a truncated form of pseudomonas exotoxin on the the plasmid vector ply5 to form the HAb25(scFv)-PE40 gene.Then the HAb25(scFv)-PE40 gene was subcloned into the pBV220 E.Coli expression vector for production of the recombinant immunotoxin.SDS-PAGE was used to identify the expression of objective protein.The specificity of HAb25(scFv)-PE40 binding to HCC cells was tested by indirect immunofluorescent staining,and the in vitro targeting cytotoxic assay was employed to observe the selective cytotoxic activity of the objective protein.Results Enzyme digestion and sequencing analysis proved the construction of HAb25(scFv)-PE40 gene in ply5 vector.HAb25(scFv)-PE40 gene was highly expressed in pBV220 vector(up to 43.3% of total cellular proteins).Indirect immunofluorescent staining showed HAb25(scFv)-PE40 could specifically bind to HCC cells as its parent antibody HAb25.The protein was verified to have selective cytotoxic activity when cocultured with HCC cells.Conclusion A single chain Fv immunotoxin HAb25(scFv)-PE40 was successfully constructed.The immunotoxin has selective cytotoxic activity against HCC,which found a basis for further study on its basic and clinical application.

Macrophage migration inhibitory factor(MIF),a potential pro inflammatory cytokine,is a pivotal regulatory element in the nature and adapted immune.This article aims to introduce the histocellular sources,genetic structure,protein structure and biological activity of this cytokine,and further to explore its role in the pathogenesis of septic shock,and its antagonist in the treatment of this disease.

In order to assess the significance of human malignancy-associated nucleolar antigen P24 in the diagnosis of tumors,552 specimens of human tissues were stained and observed with immuno - fluorescent and ABC methods using anti-P24 monoclonal antibody MA2.It was found that P24 was positive in the nucleolus of 97% cases of malignant tumors and in 50% of atypical hyperplastic tissues but negative in 98.6% cases of benign tumors and in all the normal or nontumorous tissues.On the basis of these findings,it is believed that P24 is highly specific to malignant tumors and will be a useful marker for the differential diagnosis between malignant and benign tumors.

OBJECTIVE:To establish evaluation system and quality management of dispensing department in hospital and provide basis for comprehensive and correct assessment of pharmacy dispensing work.METHODS:Based on the platform of human resource management,the flow and job responsibility of drug dispensing were adjusted."Drug dispensing work quality management specification and performance evaluation system" was established and put into practice.RESULTS:Drug dispensing quality management had criteria and principles to go by.The level of management staff were improved dramatically.58.3% of department administrators were younger than 40 years old,who were recruited and went on duty after passing examination.CONCLUSION:"Drug dispensing work quality management specification and performance evaluation system" is the comprehensive requirement of effectively ensuring the quality of drug dispending work and safety of drug utilization and reflects the standard and scientific management of drug dispensing.

Objective:To clone the cDNA in full-length of human mammaglobin,do prokaryotic expression and purify hMAM protein,as a basis for early diagnosis of breast cancer.Methods :hMAM cDNA was amplified through RT-PCR from breast cancer tissue and breast cancer cell line MD-MB453,and the recombination pQE40-hMAM vector was constructed and expressed in E.Coli.M15 after induction by IPTG.The fusion protein was purified with Ni-NTA-His affinity chromatography. Results: Two subtypes of hMAM cDNA and the hMAM(Isoform) cDNA were found,in which consisted of 270 bp,different from the wildtype hMAM cDNA of 279 bp on nine continuous base pair missing.The fusion protein formed inclusion body in prokaryotic expression system and the renatured protein was purified which purity was about 97%.Conclusion: The recombinant hMAM was successfully purified.

Objective To study absorption of shegan heji marker components in blood and their excretion in urine and feces of rats, after intragastric administration of shegan heji. Methods LC-MS/MS was used for determination of marker compounds. Rat metabolic cage technology was employed. Results Excretion of marker components were completed 24 hours after administration. Conclusion Ephedrine can be excreted from rats within 24 hours. The possibility of mutual transformation of flavonoids exists in the body. Taking shegan heji will not cause accumulation of ephedrine and flavonoids in the body.

Objective: To determine the pharmacokinetics of ephedrine hydrochloride in rats after intragastric administration of Shegan mixtures. Methods:Shegan mixtures (1. 0 ml/100 g) were administered to each rat by gavage. Blood samples were collected after the administration. Plasma concentration of ephedrine hydrochloride was determined by LC-MS/MS. The pharmacokinetic parame-ters of ephedrine hydrochloride were obtained using the pharmacokinetic software. Urine and fecal samples were collected in 24 hours after the administration using metabolic cage to determine the recovery of ephedrine hydrochloride. Results: The pharmacokinetic pa-rameters of ephedrine hydrochloride were as follows:Tmax of (1. 30 ± 0. 23)h,T1/2 of (21. 17 ± 1. 35)h, Cmax of (278. 86 ± 46. 41)ng ·ml-1,AUC0~∞ of (1221.98 ±412.64)ng·ml-1 and Vc/F of (1.70 ±0.15)L. Totally 85.66% ephedrine hydrochloride could be recovered from urine in 24 hours after the administration;however, it was not detected in the fecal samples. Conclusion: Most of e-phedrine hydrochloride is excreted through kidney in 24h,therefore, Shegan mixtures can't cause the accumulation of ephedrine hydro-chloride in rats.

Objective:To explore the efficacy and safety of transurethral anatomical vapor incision technique of prostate (VIT) with six-step method high power side-emitting greenlight laser in the treatment of benign prostatic hyperplasia (BPH).Methods:A retrospective analysis of 82 patients with BPH who used high power side-out green laser in the treatment from October 2018 to June 2020 in Gongli Hospital of Naval Medical University was performed. Among them, 40 patients were treated with six-step method VIT, and 42 patients were treated with photoselective vaporization of prostate (PVP). The two groups of patients were compared in age [(71.1±8.7)years vs.(72.1±7.0)years], prostate volume [75 (68.25, 89.00) ml vs. 73 (63.25, 85.00) ml], and peak urinary flow rate (Q max) [6.20 (5.20, 8.20) ) ml/s vs. 5.9 (4.75, 7.50) ml/s], post-void residual volume (PVR) [74.00 (42.50, 103.75) ml vs. 67.00 (58.00, 84.50) ml], international prostate symptom score (IPSS) [(21.2±5.2) vs. ( 21.0±3.9)], quality of life score (QOL) [5 (4, 6) vs. 5 (4, 6) ], prostate specific antigen (PSA) [6.20 (4.12, 8.43) ng/ml vs. 5.40 (3.88, 7.13) ng/ml ]. In general, there was no statistical difference ( P>0.05). The VIT group adopts the six-step method of marking, removing film, grooving, excision, trimming and crushing. In the PVP group, the prostate tissue was uniformly vaporized layer by layer from the inside to the outside. Perioperative indexes and complications were compared between the two groups. The Q max, IPSS, QOL, PVR and PSA between the two groups before and 3 months after surgery were compared. Results:All patients in the VIT group and PVP group successfully completed the surgery, and there was no case of transfer to TURP or open surgery. The average operation time was [60.00(50.00, 73.75)min vs. 70.00(50.00, 73.75)min] ( P<0.05). There was no significant difference in the amount of postoperative hemoglobin decline[15.00(10.00, 17.75)g/L vs. 16.00(14.00, 19.25)g/L], average bladder irrigation time[1(1, 1)d vs. 1(1, 1)d], indwelling catheterization time[3(3, 3)]d vs. 3(3, 3)d] and hospitalization time in patients after operation[4(3, 4)d vs. 4(4, 4)d] ( P>0.05). All patients had no blood transfusion, second bleeding, readmission, TURS, urethral stricture and urinary incontinence.There were 2 cases (5.0%) of postoperative urinary tract infection in the VIT group and 9 cases (21.4%) of postoperative urinary tract infection in the PVP group ( P<0.05), and they were cured after anti-inflammatory treatment. Three months after operation, Q max, IPSS, QOL, PVR and PSA in the two groups were significantly improved compared with preoperatively. Among them, the differences of IPSS [(5.7±2.5) points vs. (7.5±2.8) points] and PSA [2.65(2.10, 3.90)ng/ml vs. 4.00(2.45, 4.45)ng/ml] in the VIT group and PVP group after operation were statistically significant ( P<0.05). Conclusions:Applying the six-step method high power side-emitting greenlight laser transurethral anatomical VIT to treat BPH, there is less intraoperative and postoperative bleeding, short operation time, significant decrease in PSA, and fewer complications. It is a safe and effective minimally invasive technology for the treatment of BPH.