Objective:Evaluation of the effectiveness of domestic ultrapulsed thulium laser ablation of isolated porcine kidneys.Methods:Forty-two fresh porcine kidneys were taken. One porcine kidney was dissected along the sagittal plane at the renal hilum, and then dissected along the coronal plane and divided into four pieces.A total of 168 pieces of renal tissues were obtained, of which 162 pieces were selected for the domestic hyperpulsed thulium laser ablation of isolated porcine kidneys using the single-needle method and the double-needle method, respectively. The ablation was carried out with different ablation modes (single-needle and double-needle methods), ablation power (4, 5, and 6 W), ablation modes (continuous, low-energy and high-frequency mode, and high-energy and low-frequency mode), and ablation time (10, 15, and 20 s). The size of the ablation area in terms of the long diameter, the short diameter, and the thick diameter were measured and recorded. The embedded tissues were sectioned and stained with HE to study the microscopic pathological characteristics of the ablation foci. The ablation foci volume and sphericity coefficient (Φ) were calculated, and the ablation foci volume and sphericity coefficient were statistically analyzed by analytic factorial design and one-way ANOVA.Results:The gross specimen showed that the single-needle ablation foci were elliptical in shape, containing black carbonized areas and gray-white necrotic areas with clear boundaries, and the peripheral renal tissues were pink, and the ablation focus was clearly demarcated from the normal tissues. The double-needle ablation foci were a large ellipse containing two small ellipse ablation foci, and the structure was similar to that of the single-needle method. Under the light microscope, the center of the ablation focus was a "cavity-like" carbonized area, outside of which was an area of coagulation necrosis, with deepened cell staining, reduced cell volume, disorganized arrangement, loose nuclei, nuclear consolidation, and some cell nuclei were cleaved and ablated. Ablation mode ( P<0.001), ablation pattern ( P <0.001), ablation time ( P <0.001), and ablation power ( P <0.001) all significantly affected ablation focus volume. The volume of the ablation foci increased with increasing number of needle feeds, longer ablation time, higher ablation power, and with the ablation mode being continuous mode. Single-needle ablation produced the largest volume of ablation foci (259.56 mm 3) in continuous mode, ablation time of 20 s, and ablation power of 6 W. Double-needle ablation produced the largest volume of ablation foci (452.00 mm 3) in continuous mode, ablation time of 20 s, and ablation power of 6 W. The analysis of the sphericity coefficient showed that in single-needle ablation, the sphericity coefficient was the largest (Φ=0.76) under the conditions of continuous mode, ablation power of 4 W, and ablation time of 15 s. The sphericity coefficient was the smallest (Φ=0.56) under the conditions of high-energy, low-frequency mode, ablation time of 10 s, and ablation power of 6 W, and the morphology of the ablation foci were more close to ellipsoid shape. When ablated by the double-needle method, the ablation foci morphology was closer to spherical shape (Φ=0.91) under the conditions of continuous mode, ablation time of 10 s, and ablation power of 5 W. The combined main effect and interaction results showed that ablation mode ( P<0.001), ablation pattern ( P<0.001), and ablation power ( P<0.001) significantly affected the sphericity coefficient of the ablation foci, and ablation time did not affect the ablation foci morphology. Conclusions:Domestic ultrapulsed thulium laser ablates isolated porcine kidney tissues with precise effect, the tissue morphology of the unablated area is normal, the thulium laser ablated foci have a definite safe boundary. Choosing double needle method, continuous mode, ablation power 6 W for 20 s, can ablate spherical cancer foci within 1 cm in diameter, which can be used for spherical small kidney cancer ablation. Selecting the single-needle method, high-frequency, low-energy mode and ablation power of 6 W, the ablation foci were similar to the oval volume, and could be used for the ablation of oval small renal cancer. However, this study is only limited to the animal ex vivo model, which needs to be confirmed by further study.

Objective:To develop the discrimination experience questionnaire for HIV/acquired immunodeficiency syndrome(AIDS) patients and test the reliability and validity of the questionnaire.Methods:Based on the literature review and semi-structured interviews to clarify the operational definition of discrimination for HIV/AIDS and develop the item pool. The questionnaire was developed though 2 rounds Delphi consultation and a pilot test. A total of 410 HIV/AIDS patients in Shanghai Public Health Clinical Center of Fudan University from June to December 2020 were selected to investigate the questionnaire, item analysis was used to screen items. SPSS 22.0 software was used for reliability test and exploratory factor analysis, the AMOS 21.0 software was used for confirmatory factor analysis to test the reliability and validity of the questionnaire.Results:The questionnaire consisted 2 dimensions(external discrimination and internal discrimination) and 10 items. Exploratory factor analysis showed that two common factors were extracted from the frequency of discrimination and the degree of negative psychological impact of discrimination experience on patients, and the cumulative variance contribution rates were 48.367% and 55.403%, respectively. The confirmatory factor analysis on the frequency of discrimination showed that Chi square degree of freedom ratio ( χ2/ df) was 2.831, P<0.05, root mean square of approximation error (RMSEA) was 0.093, goodness of fit index (GFI) was 0.928, comparative fit index (CFI) was 0.925, incremental fit index (IFI) was 0.926; the confirmatory factor analysis on the negative psychological impact of discrimination experience on patients showed that χ2/ df was 1.740, P<0.05; RMSEA was 0.076, GFI was 0.925, CFI was 0.936, IFI was 0.938. The content validity of the questionnaire was 0.9. The Cronbach α coefficientof questionnaire was 0.811, and the test-retest coefficient was 0.862 ( P<0.01). Conclusions:The discrimination experience questionnaire for HIV/AIDS patients has good reliability and validity, and it can be used to measure the discrimination for HIV/AIDS patients.

Vaccination is the most effective way to prevent influenza and severe outcoming caused by influenza viruses.Health care workers(HCW) are exposed to patients with influenza and they are at high risk of occupationally acquired influenza and of causing nosocomial infection among patients,increasing the incidence rate,the risk of severe and death of patients.Improving the influenza immunization in HCW can not only reduce the prevalence of themselves and keep a weel-oiled of health care facilities during the influenza seasons,but also reduce the risk of severe and death among patients and increase the influenza vaccine uptake in whole population.At present,the influenza immunization coverage of HCW is low.The obstacles and myths of influenza vaccine are barriers for vaccine uptake among HCW.The various strategies are critical in order to improve the influenza coverage rates of HCW.

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Objective: To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression. Methods: UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by ³²P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager. Results: UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA. Conclusions UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.

Objective To investigate the incidence and risk factors of acute kidney injury (AKI)in hepatitis B virus (HBV)related acute-on-chronic liver failure (ACLF)patients,and to explore the impact of AKI on the prognosis of ACLF.Methods The medical records of 227 patients who were diagnosed with HBV-related ACLF at the Department of Infectious Diseases in the First Affiliated Hospital of Nanchang University from January 2015 to August 2016 were retrospectively reviewed.Patients were divided into AKI group and non-AKI group based on the AKI criteria published by International Club of Ascites in 2015 .Demographic and clinical data were compared between groups.The AKI incidence and its impact on patients’prognosis were analyzed.The comparison of continuous variables was done by t test or rank-sum test.The comparison of categorical variables was done byχ2 test or Fisher exact test.AKI risk factors were analyzed by using logistic regression.Results There were 66 (29.1 %)cases were diagnosed with AKI among 227 ACLF patients,among which,45 patients (68.2%)were stage Ⅰ,14 (21 .2%) were stage Ⅱ and 7 (10.6%)were stage Ⅲ.Age,cirrhosis,concentrations of total bilirubin and albumin,international normalized ratio (INR),percentage of neutrophils,MELD scores and spontaneous peritonitis rate (SBP)were all statistically different between AKI group and non-AKI group (all P <0.05).The binary logistic regression analysis revealed that only INR (OR=3.132,P =0.001 )and SBP (OR=4.204,P =0.001 )were the independent risk factors of AKI.The optimal cut-off value for INR was 2.025 with AUROC of 0.609 (P =0.01),sensitivity of 59.1 % and specificity of 62.1 %.The 30-day mortality of AKI group was significantly higher than non-AKI group (χ2= 18.324,P < 0.01). Conclusions AKI is relatively common in patients with ACLF.The risk factors of AKI are INR and SBP. AKI has significant impact on the short-term survival rate of ACLF.Therefore,physicians should pay attention to patients with INR of ACLF at admissions and SBP during the management so as to prevent the occurrence of AKI and to reduce the fatality of ACLF.

Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.

Objective To study the influence of Annao tablet on the expression of transforming growth factor beta 1 (TGF-β1) in acute graft-versus-host disease (aGVHD) murine and to explore the interventional mechanism of TGF-β1 on aGVHD. Methods Hematopoietic stem cells of male Balb/c mice were transplanted to female C57BL/6 mice for the development of aGVHD murine model. Recipient mice were divided into Annao group and blank group randomly and respectively administrated with Annao soup (a kind of Chinese herb) and 0.9% sodium chloride intragastrically. Clinical symptom, survival time and body weight were recorded at 14th and 30th day and some sections of liver, small bowel and skin were taken for histological changes. Serum level of TGF-β1 were measured by enzyme-labeled immunosorbent assay (ELISA), splenocyte protein of TGF-β1 by Western Blot and TGF-β1 mRNA by fluorescent quantitation polymerase chain reaction (PCR). Results Serum level of TGF-β1 in both groups had no statistical difference (P = 0.305), but it rose to (148.31 ± 7.95) ng/mL at 14th day and (183.48 ± 5.91) ng/mL at 30th day in Annao group, which had significant difference when compared with that in blank group (P = 0.000). IOD/IODβ-actin value of TGF-β1 protein in Annao group was 0.33 ± 0.05 at 14th day and 0.56 ± 0.04 at 30th day, which was higher than that in blank group (P = 0.000) and the expression of TGF-β1 mRNA of splenocyte in Annao group was 1.24 ± 0.04 at 14th day and 2.14 ± 0.33 at 30th day which was much higher than that in blank group (P = 0.000). Conclusion Annao tablet helps to relieve symptoms of acute GVHD by raising serum level of TGF-β1 and intensifying expression of protein of TGF-β1 and its mRNA.

Objective To investigate the therapeutic effects of haploidentical hematopoietic stem-cell transplantation (Haplo-PBSCT) for acute myeloid leukemia in first relapse after complete remission by standard induction chemotherapy. Methods Eighty-nine cases of AML in first relapse after complete remission by standard DA/Hi-Ara-C regimens induction chemotherapy were evaluated retrospectively. Fiftythree cases were grafted by haplo-PBSCT and 26 cases were treated with iDA/Mid-Ara-C or MA/ Mid- Ara-C agents. Results The second remission rate in haplo-PBSCT group and continuous chemotherapy group was 86. 7 % (46/53 cases) and 38. 1% (9/23 cases) respectively (P＜0. 01). Survival postprogression (SPP) at 36th month was 43. 4 % (23/53 cases) in haplo-PBSCT group and 11.5 % (3/26 cases) in continuous chemotherapy group (P ＜ 0. 05). Conclusion Haplo-PBSCT could significantly increase the second remission rate and prolong the survival time of patients with acute myeloid leukernia in first relapse after complete remission by standard induction chemotherapy.

Objective A group of patients who had undergone total hip arthroplasty were followed up radiologically to explore the effects of abduction angle, the anteversion angle and the height of the femoral head center on dislocation, because these measurements could be used as a basis to evaluate current practice and to seek improvements. Methods A series of 326 hip joints (318 patients) were followed up routinely. The average follow up period was 2.7 years. 10 was found to have dislocation. The abduction angle, the anteversion angle and the height of the femoral head center were measured. The results were statistically analyzed, with a P value less than 0.05 indicating significant difference. Results The abduction angle of 55?or larger was found to be associated with greater risk of dislocation, compared with the abduction angle of less than 55?. Meanwhile, cups with the femoral head center superior placement greater than 30 mm resulted in more dislocation, compared with those less than 30 mm. As an independent variable, the anteversion angle had no significant association with dislocation. Conclusions Based on the results of the current study, the abduction angle and the height of the femoral head center should be considered as prerequisites for maintaining hip stability and decreasing the risk of dislocation. We believe that hips with the abduction angle of less than 55?and the height of the femoral head center less than 30 mm can decrease the risk of dislocation.