Objective To express and purify the protein of variable region of adeno-associated virus 9(AAV9) capsid in prokaryotic cells,and prepare rabbit polyclonal antibody against it.Methods DNA sequence encoding variable region of AAV9 capsid protein was designed,synthesized and inserted into prokaryotic expression vector pET-30a. The obtained plasmid pET-30a-AAV-VR was transformed to E.coli BL21(DE3),induced by IPTG to express the multivalent antigenic peptide and purified by Ni-NTA resin under denaturation conditions. Male Japanese large-eared white rabbits were immunized with the AAV9 variable region protein after dialysis and renaturation to prepare polyclonal antibody,which was determined for the antibody potency by indirect ELISA,and for the specificity by Western blot and cellular immunofluorescence.Results The recombinant prokaryotic expression plasmid pET-30a-AAV-VR expressing the variable region of AAV9 capsid was constructed correctly as identified by XhoⅠ and BglⅡ digestion. The expressed protein was recognized by His tag antibody after purification with a relative molecular mass of about 20 000. The potency of rabbit polyclonal antibody was 1∶10 240 000,which specifically recognize AAV9 capsid protein.Conclusion The capsid variable region protein of AAV9 was successfully expressed and rabbit polyclonal antibody with high potency was prepared,which laid a foundation of the subsequent development of AAV vector and the research of AAV biological function.

The rapid detection technology in food safety plays a vital role in the field of preventive medicine.The traditional physical and chemical detection methods have some limitations,such as costly,difficult to operate and high requirements on environment,can not meet the needs of quality and safety on-site rapid screening,and are gradually replaced by emerging sensor detection technologies.The aptamer-upconversion biosensor technology is a novel rapid detection technology,which is based on the combination of functionalized upconversion nanomaterials and aptamer technology.Compared with the traditional immunofluorescence technology,it has advantages in sensitivity,specificity and stability,and is widely used in rapid detection.In this paper,the upconversion luminescence and aptamer technologies were briefly introduced,and the aptamer-based upconverting phosphor biosensing technology and its application in rapid detection of food safety were summarized.Based on the current research status,the bottleneck and the future development trend of this technology were analyzed,which provides a reference for the application of this technology in food safety and other fields.

Objective:To explore the diagnostic value of 68Ga-prostate specific membrane antigen (PSMA)-11 PET/MR multiparameter multimodal functional imaging in the diagnosis of naive prostate cancer (PCa), and to analyze its efficacy in the early stages of PCa. Methods:From July to September 2019, 45 suspected or pathologically confirmed PCa patients (average age: 69 years) who met the inclusion criteria were collected to perform 68Ga-PSMA-11 PET/MR examination in Nanjing First Hospital. After the scanning was completed, the method of region of interest (ROI) was used to semi-quantitatively calculate the tumor radioactive uptake in the fusion image, including the maximum standardized uptake value (SUV max), tumor metabolic volume (MTV) and mean standardized uptake value (SUV mean), and PSMA expression load was calculated(SUV mean×MTV). Apparent diffusion coefficient (ADC) values of ROI were measured in ADC images ( b=1 500 s/mm 2). The efficacy of 68Ga-PSMA-11 PET/MR in the preoperative diagnosis of PCa and the effect on clinical staging were evaluated with the pathological results. The correlation between prostate specific antigen (PSA) and radiation uptake in PCa tissues, PSMA expression load and ADC values was analyzed by Pearson correlation. Independent-sample t test was used to analyze the data. Results:Pathologically, 38 of the 45 patients were with PCa and 7 patients had benign prostatic hypertrophy (BPH), and metastasis occurred in 12 of the 38 PCa patients. 68Ga-PSMA-11 PET/MR detected 39 cases of PCa with one of which was false-positive. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of 68Ga-PSMA-11 PET/MR were 100%(38/38), 6/7, 97.4%(38/39), 6/6 and 97.8%(44/45), respectively. The tumor tissues of PCa often showed focal radioactive uptake, and the T 2 weighted imaging (WI) showed focal low signal and limited dispersion. BPH showed slightly uneven uptake, and T 2WI and diffusion weighted imaging (DWI) showed uneven diffuse signals. SUV max of PCa was significantly higher than that of BPH (24.66±19.21 vs 4.97±2.13; t=5.208, P<0.001). ADC values of PCa were significantly lower than that of BPH ((0.91±0.37)×10 -3vs (1.08±0.24)×10 -3 mm/s 2; t=2.816, P<0.05). SUV max and the expression loads of PSMA in PCa were positively correlated with PSA ( r values: 0.42 and 0.71, both P<0.05). ADC values of tumor tissues in PCa were negatively correlated with PSA ( r=-0.37, P=0.013). Conclusion:68Ga-PSMA-11 PET/MR has great merits in the early diagnosis and staging of PCa.

OBJECTIVE:To optimize the extraction technology of Baiyi rectal solution. METHODS:The effects of water amount and extraction time on extraction technology were investigated by central composite design-response surface method using OD value of berberine hydrochloride and ferulic acid and the extract yield as index. Validation test was also conducted. RESULTS:The optimal extraction technology was as 11-fold water,decocting 70 min for the first time;5-fold water,decocting 30 min for the second time. In validation test,OD value of first extraction was 0.940 0,and the bias between observed and predicted values was -2.07%;that of second extraction was 0.851 8,and the bias was -2.41%. CONCLUSIONS:The extraction technology of Baiyi rectal solution is reasonable,feasible and suitable for industrial production.

Objective:To understand the laboratory characteristics for the diagnosis of immunoglobulin G4-related disease (IgG4-RD).Methods:The clinical data of 28 patients with IgG4-RD and renal damage (IgG4-RKD) diagnosed in our hospital from January 2017 to May 2019 were retrospectively analyzed. The correlation between serum IgG4 concentration and clinical features as well laboratory test results was analyzed. The 28 patients were divided into two groups: high serum IgG4 concentration group and normal serum IgG4 concentration group. The serum creatinine value, erythrocyte sedimentation rate, IgG concentration, IgA concentration, complement C3, C4 concentration, peripheral blood eosinophils, hemoglobin, IgG4/IgG and other related parameters were compared between the two groups. SPSS 20.0 statistical software was used for analysis. The two groups of measurement parameters were compared between groups by independent sample t test, non-normal measurement parameters were compared between groups by Mann-Whitney U test analysis, and the correlation between patients' IgG4 and each detection parameter was analyzed by Spearman correlation analysis. Results:Among the 28 patients, 17 were male and 11 were female, with an average age of (62±14) years. The serum IgG4 concentration increased in 75% of the patients ( n=21), with an average value of 3.01(1.41, 7.52) g/L, the serum IgG concentration increased in 64.3% of patients ( n=18), with an average value of 18.91 (12.88, 24.88) g/L, and the complement C3 decreased in 50% of the patients ( n=14), with an average value of(0.77±0.28) g/L. IgG4 was positively correlated with IgG ( r=0.422, P=0.025), IgG4/IgG ( r=0.951, P<0.01), ESR ( r=0.543, P<0.01) and peripheral blood eosinophils ( r=0.487, P<0.01), but negatively correlated with complement C3 ( r=-0.431, P=0.022) and C4 ( r=-0.504, P<0.01) levels. There were significant differences in IgG ( Z=-2.255, P=0.023), IgG4/IgG ( Z=-3.793, P<0.01), C3 ( t=7.380, P<0.01) and ESR ( t=-2.195, P=0.037) between the elevated IgG4 group and the normal group. Conclusion:Serological characteristics of IgG4-RKD combined with clinical manifestations may be able to diagnose IgG4-RKD in early stage.

Objective To evaluate the efficacy of Disposcope endoscope for tracheal intubation in patients with difficult glottis exposure.Methods Sixty adult patients of both sexes,aged 22-68 yr,scheduled for elective oro-tracheal intubation under general anesthesia,whose glottis was not visible under Macintosh laryngoscope (grade Ⅲ or according to Cormach-Lehane Grading of laryngoscopic view),were randomized into 2 groups (n =30 each) using a random number table:group Macintosh laryngoscope-assisted Disposcope endoscope (group DM) and group Disposcope endoscope (group D).The glottis exposure condition and time,intubation condition and time,assisted lifting mandible,laryngeal mucosa bleeding and postoperative sore throat and hoarseness were recorded.Results The success rate of intubation at second attempt achieved 100％ in the two groups.Compared with group DM,the incidence of assisted lifting mandible was decreased,and no significant changes were found in the success rate of glottis exposure,glottis exposure time,intubation time,success rate of intubation at first attempt,incidence of laryngeal mucosa bleeding,and postoperative sore throat and hoarseness in group D.Conclusion Disposcope endoscope can improve the condition for tracheal intubation significantly after lifting mandible in patients with difficult glottis exposure.

Objective To further control the indications and to improve the effect of intra-esophageal metallic stents.Methods 83 metallic stents were be planted into esophageal through endoscopy and under X-ray TV guiding using two kinds of stent meterials in 72 cases with benign or maligant stenosis(esophageal,cardiac orifice).Results The operation of planting stents was successful in all patients and the effect of treatment of esophageal stenosis with stents covered by dacron kint membrane was best.The clinical symptoms and food intaking ablities of all patients were improved.Conclusion Home-made nitinol stents can be plant in treatment of malighent esophageal stenosis.It should be first choice for the patients without indications of surgery or after operation.

Objective: Our aim was to study the role of telomerase activation in the course of cervical carcinogenesis and progression.Methods:Telomeric repeat amplification protocol(TRAP) assay was used to measure telomerase activity in tissue samples with various cervical conditions:40 with cervical cancer, 50 with cervical intraepithelial neoplasia(CIN), 20 with normal cervice. Results:The positive rate of telomerase activity was 95.0%,44.0%, and 10.0% in cervical cancer, CIN, and normal cervices, respectively, which was significantly higher in cervical cancer than that in CIN and normal cervices, so was that in CIN than that in normal cervices (P<0.01) . The positive rate was 22.2%, 37.5%, and 75.0% in CINⅠ,Ⅱ, and Ⅲ, respectively, which was significantly higher in CINⅢ than that in CIN Ⅱand CINⅠ (P<0.01).Conclusion:Telomerase activation may relate to cervical carcinogenesis, which correlates well with the grade of cervical lesions.

Objective To investigate the relationship between cytochrome P450 1A2 (CYP1A2) gene polymorphism and susceptibility to chronic obstructive pulmonary disease (COPD). Methods CYP1A2 gene polymorphisms in 100 COPD cases and 100 healthy controls were tested with polymerase chain reaction- restriction fragment length polymorphism (PCR- RELP). Results Genotype frequencies of 4 SNPs in both the COPD and control groups were in accordance with Hardy-Weinberg equilibrium (P>0.05). There was significant difference between the COPD and control groups in genotype and allele frequencies of 1D and 1F (P<0.05), but not of 1C and 1E (P>0.05). Conclusion CYP1A2*1D and CYP1A2*1F polymorphisms may play an important role in the development of COPD.

Objective To study expression,distribution,significance and relationship of CXCL12,CXCR4,microvessel density (MVD) in multiple myeloma (MM) bone marrow micro niche.Methods 63 cases of patients with MM were chosen as the experimental group,42 cases of healthy persons were chosen as the control group.The expression and distribution of CXCL12,CXCR4,MVD were detected by immunohistochemical method.Results Compared with the control group,the expressions of CXCL12,CXCR4,MVD in the experimental group were higher,the difference was statistically significant (P ＜ 0.05),their expressions in bone marrow were not significantly different from gender,age,immunoglobulin type and light chain classification (P＞0.05);the expression of CXCR4,MVD were positively related with CXCL12 (P＜0.05).Conclusion The expression of CXCL12,CXCR4,MVD may be related to the occurrence of MM;CXCL12/CXCR4 biological axis could promote the angiogenesis of MM bone marrow micro niche.