Objective:We genetically modified our non-replicating vaccinia virus NTV to improve its immunogenicity.Methods:We constructed NTV-modified strain NTV-ΔF1L-C7L by homologous recombination of vaccinia virus based on CRISPR-Cas9 technology by inserting the C7L gene while deleting the F1L gene. The recombinant virus NTV-ΔF1L-C7L was then immunized with 10 7 PFU in BALB/c mice, and the levels of humoral and cellular immunity induced by NTV-ΔF1L-C7L were detected by ELISA and ELISpot method, respectively, and the levels of neutralizing antibodies were determined by the phage-reduced neutralization assay. Results:The PCR and western- blot identification proved that the F1L gene of the constructed NTV-modified strain NTV-ΔF1L-C7L was missing, while the C7L gene was inserted back in the region, and the C7L gene could be expressed normally, indicating that the recombinant virus was constructed correctly. After immunization of mice with NTV-ΔF1L-C7L, ELISA result showed that the recombinant virus NTV-ΔF1L-C7L induced a higher level of IgG antibody than NTV; ELISpot result also showed that the recombinant virus was able to induce a higher level of IFN-γ; and the result of plaque reduction neutralization test showed that the recombinant virus was able to induce a higher level of IFN-γ antibody than that of NTV.Conclusions:We correctly constructed the NTV gene-modified strain NTV-ΔF1L-C7L, which induced stronger humoral and cellular immunity compared with NTV, and provided reference data for the research and development of replacement products for smallpox or monkeypox vaccines.

TCM dispensing is the most basic clinical pharmaceutical work of TCM.In recent years,based on the 9 key technologies of TCM dispensing,the TCM dispensing industry has ushered in great development,and innovative TCM dispensing information system and intelligent dispensing equipment have appeared.This article sorted out the current situation of TCM dispensing industry and looked forward to its future development route.The results showed that the introduction of new technology and new equipment in the key technical links of procurement acceptance,dispensing review,TCM decocting,medication guidance and so on have improved the quality of dispensing service and ensured the quality and safety of medication.In the development of modern TCM dispensing industry,it is necessary to improve the quality control standard system,service standard system and core equipment standard system in the standardization of dispensing technology;in terms of talent cultivation in the field of dispensing,it is necessary to focus on restructuring and building new educational models to cultivate new medical talents that intersect medical and engineering fields;in terms of informatization and intelligence,it is necessary to develop intelligent equipment that is more in line with the characteristics of TCM,and further promote and improve the"shared TCM pharmacy"model.Through improving the content of TCM clinical pharmaceutical care,developing new technology and equipment of TCM dispensing,and improving the level of dispensing service and education,it is expected to gradually realize the standardization,informatization and intelligent development of modern TCM dispensing industry.

Objective:To evaluate the functional and anatomical outcomes of autologous single retinal pigment epithelium (RPE) transplantation for severe obsolete submacular hemorrhage (SMH) in late age-related macular degeneration (AMD).Methods:A retrospective clinical study. From January 2012 to December 2015, 11 patients with AMD (11 eyes) with obsolete SMH who were diagnosed and treated by pars plana vitrectomy (PPV) combined with autologous RPE transplantation at the Department of Ophthalmology, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were included. Among them, there were 9 eyes in 9 males and 2 eyes in 2 females. All the eyes underwent the examinations of best corrected visual acuity (BCVA) and optical coherence tomography; 4 eyes underwent macular fixation function (MAIA) at the same time. The BCVA examination was carried out using the international standard visual acuity chart, which was converted into logarithm of the minimum angle of resolution (logMAR) visual acuity during statistics. All eyes were treated with PPV combined with autologous single-layer RPE transplantation or autologous RPE-choroidal full-thickness transplantation, and were divided into S group and C group, with 5 and 6 eyes respectively. The differences of age ( t=-0.363), gender composition ratio ( χ2=0.549), course and thickness of SMH ( t=0.118, 0.231), average times of anti-vascular endothelial growth factor drug treatments ( t=0.129), times of PPV ( t=-0.452) between the two groups were not statistically significant ( P>0.05). The follow-up period was 6-40 months after the operation, and the BCVA, MAIA, graft status and complications of the eyes after the operation were observed. The comparison of continuous variables between groups was performed by independent-sample t test; the comparison of categorical variables was performed by χ2 test. Results:At the last follow-up, the average logMAR BCVA of the eyes in group S and C were 1.62±0.34 and 1.03±0.20, respectively; group C was better than group S, however, the difference was not statistically significant ( t=1.532, P=0.160). There were 4 eyes (80%, 4/5) and 6 eyes (100%, 6/6) in S group and C group with BCVA better than preoperative, the difference was no statistical significance ( χ2=0.677, P=0.895). There were 2 (40%, 2/5) and 3 (50%, 3/6) eyes with logMAR BCVA better than 1.0 in S group and C group, and the difference was not statistically significant ( χ2=0.572, P=0.423). After the operation, 6 eyes of grafts were in good condition and 5 eyes were in poor condition; the BCVA of grafts in good condition was significantly higher than that of poor condition, the difference was statistically significant ( t=4.894, P=0.001). Among the 4 eyes that underwent MAIA examination, 2 eyes were unstable and diffusely fixed on the graft; the fixation point was located at the normal retina adjacent to the graft area in 2 eyes. Secondary subretinal hemorrhage occurred in 3 eyes after the operation; the intraocular pressure was high in 1 eye after the operation. During the follow-up period, no intraocular infection, secondary retinal detachment, recurrent choroidal neovascularization or low intraocular pressure occurred in all eyes. Conclusions:Both autologous single-layer RPE transplantation and autologous RPE-choroidal full-thickness transplantation can help stabilize or even improve the visual function of eyes with severe SMH secondary to advanced AMD. The visual acuity after surgery is closely related to the state of the graft.

Objective:To investigate the efficacy and safety of humanized CD19-specific chimeric antigen receptor T cells (hCART19s) in treating children and young adults with relapsed/refractory acute lymphoblastic leukemia (R/R ALL) and to analyze relevant factors affecting its curative effect and prognosis.Methods:We conducted a single-center clinical trial involving 31 children and young adult patients with R/R B-ALL who were treated with humanized CD19-specific CAR-T cells (hCART19s) from May 2016 to September 2021.Results:Results showed that 27 (87.1%) patients achieved complete remission (CR) or CR with incomplete count recovery (CRi) one month after CAR-T cell infusion. During treatment, 20 (64.5%) patients developed grade 1-2 cytokine release syndrome (CRS) , and 4 (12.9%) developed grade 3 CRS. Additionally, two patients had grade 1 neurological events. During the follow-up with a median time of 19.3 months, the median event-free survival (EFS) was 15.7 months (95% CI 8.7-22.5) , and the median overall survival (OS) was 32.2 months (95% CI 10.6-53.9) . EFS and OS rates were higher in patients who have undergone hemopoietic stem cell transplantation (HSCT) than in those without [EFS: (75.0 ± 12.5) % vs (21.1 ± 9.4) %, P=0.012; OS: (75.0 ± 12.5) % vs (24.6 ± 10.2) %, P=0.035]. The EFS and OS rates were significantly lower in patients with >3 treatment lines than in those with <3 treatment lines [EFS: 0 vs (49.5±10.4) %, P<0.001; OS: 0 vs (52.0±10.8) %, P<0.001]. To the cutoff date, 12 patients presented with CD19 + relapse, and 1 had CD19 - relapse. Conclusion:hCART19s are effective in treating pediatric and young adult R/R ALL patients, with a low incidence of severe adverse events and reversible symptoms. Following HSCT, the number of treatment lines can affect the long-term efficacy and prognosis of pediatric and young adult R/R ALL patients.

Xp11.2 translocation/TFE3 gene fusion-related renal cancer often affects young people, and lymph node metastasis is also common, with a good short-term prognosis. This article reports a 19-year-old young female with a huge metastatic Xp11.2 translocation/TFE3 gene fusion-related renal carcinoma, which metastasize to lung and lymph node. The patient underwent axitinib neoadjuvant targeted therapy before the operation. During the operation, lymph node metastasis was found and the para-aortic lymph node dissection was performed.After the operation, the patient continued to be treated with axitinib, and the lung metastases improved. There was no local recurrence and metastasis after 1 year follow-up.

Objective:To investigate the relationship between genotype and phenotype in children with CRB1 mutated Leber congenital amaurosis (LCA) and early onset retinal dystrophy (EOSRD). Methods:A retrospective clinical study. From January 2013 to December 2019, 10 children with CRB1 mutated LCA/EOSRD were enrolled in the study. The patients were identified as CRB1 mutation by the second generation targeted capture sequencing, Sanger sequencing and the family segregation analysis. All children underwent electroretinogram (ERG) and fundus examination. At the same time, 6 cases were examined by optical coherence tomography (OCT); 1 case was examined by fluorescein fundus angiography (FFA), 7 cases were examined by wide-angle laser scanning ophthalmoscope (UWF SLO). Results:There were 6 cases of LCA and 4 cases of EOSRD in 10 patients with CRB1 gene mutations. The average age of first visit was 3.61 years old. The light and dark wave of ERG was flat in 6 cases, and decreased in 4 cases. A total of 19 pathogenic mutations were detected. There were 1 homozygous mutation and 9 compound heterozygous mutations. There were 4, 2 and 1 cases of "copper-coin" like, "salt and pepper" like and "osteocyte" like pigment changes in retina, 1 case of "crystalline pigment" change and 2 cases of macular pigment scar. In 7 cases of UWF SLO examination, different degrees of para-arteriolar pigment epithelium retention (PPRPE) were found in the middle and peripheral fundus. In 6 cases examined by OCT, the outer layer of retina atrophied and the band of ellipsoid disappeared. Symmetrical cystoid macular edema, splitting cystoid macular degeneration and adhesion of epi-macular membrane to optic disc and macular area were found in 1 case, respectively, the retinal structure was rough and thickened, and the fovea became thinner in 3 cases. In FFA examination, 1 case showed uveitis-like changes with late optic disc fluorescein staining, macular fluorescence accumulation, strong fluorescence diffusing along the blood vessels in each quadrant, peripheral PPRPE of "frost-branch" like strong fluorescence. Conclusion:The relationship between genotype and phenotype of CRB1 mutation is complex, and PPRPE is a common characteristic change.

The urothelium is a transitional epithelium which lines on the renal pelvis, ureter, bladder and proximal urethra adjacent to the bladder. It’s a permeability barrier. In the process of urothelial development or injury repair, the mature urothelial cells expressing uroplakin appears after the hierarchical transcription of a series of transcription factors in basal stem cells. Understanding normal urothelial differentiation process of the urothelial cells could be beneficial to uncover the development of urothelial carcinoma so that to provide targeted therapy options and the study of the urinary tract repair process after injury. In addition, the study of stem cell differentiation microenvironment also provides important theoretical support for tissue engineering and tissue reconstruction. In this review, we discuss the latest findings on urothelial cell differentiation and its clinical significance.

Objective:To evaluate the prognostic value of combined detection of serum C-reactive protein(CRP), procalcitonin and lactic acid in elderly patients with community acquired pneumonia(CAP).Methods:Ninety-five elderly CAP patients in the emergency department of Shangqiu First People's Hospital were included as the case group, and 45 elderly healthy people in the emergency department of Shangqiu First People's Hospital were enrolled as the control group.Levels of blood lactic acid, procalcitonin and CRP were compared between the two groups.Meanwhile, blood levels of lactic acid, procalcitonin and CRP were compared between patients with different outcomes in the case group.The receiver operating characteristic(ROC)curve was used to evaluate the prognostic value of CRP, procalcitonin and blood lactic acid in elderly patients with CAP.Results:Compared with the control group, blood levels of lactic acid, procalcitonin and CRP were increased in the case group( t=20.77, 26.03 and 31.27, all P<0.01). During a 12-month follow-up, 13 cases(13.68%, 13/95)died and 82 cases(86.32%, 82/95)survived in the case group.Blood levels of lactic acid, procalcitonin and CRP were higher in the death group than in the survival group( t=25.56, 8.30 and 13.56, all P<0.01). ROC curve analysis showed that the sensitivity and specificity of serum lactic acid, procalcitonin and CRP in predicting the prognosis of elderly CAP patients were 76.92% and 73.17%, 84.62% and 78.05%, and 69.23% and 70.73%, respectively.The sensitivity and specificity of combined detection of the three indicators were 92.31% and 89.02%, respectively, higher than those of the individual indicators. Conclusions:Serum levels of lactic acid, procalcitonin and CRP are increased in elderly patients with CAP.Combined detection of the three indicators can improve the prognostic value and therefore has important clinical significance.

OBJECTIVETo investigate the role of poly(ADP-ribose) polymerases-1 (PARP-1)-mediated blockade of autophagic flow in myocardial ischemia-reperfusion injury.

METHODSH9c2 cells, a rat cardiac myocyte line, were divided into control group, hypoxia/ reoxygenation model group (H/R group), PARP-1 inhibitor (PJ34) group, and PJ34 + H/R group. The total protein was extracted from the cells in each group to detect the expressions of pADPr, Bax, the DNA damage marker protein p-YH2ax, and autophagic flow-associated proteins LC3BⅡ/LC3Ⅰ, Beclin-1, and P62 using Western blotting.

RESULTSCompared with the control cells, the cells with H/R exhibited significantly increased expressions of pADPr, Bax and p-YH2ax ( < 0.05). The expressions of LC3B Ⅱ, beclin-1 and p62 were also increased significantly in the cells with H/R ( < 0.05), indicating the block of the autophagic flow. The application of PARP-1 inhibitor PJ34 in the cells with H/R significantly inhibited the expressions of pADPr ( < 0.05) and Bax ( < 0.01), and alleviated DNA damage in the cells. PJ34 treatment did not cause significant changes in the expressions of LC3B Ⅱ and beclin-1 but significantly decreased the expression of p62 ( < 0.05) in the cells with H/R.

CONCLUSIONSBlock of autophagic flow mediated by PARP-1 activation plays a role in myocardial ischemiareperfusion injury, and inhibition of PARP-1 activity can reverse autophagic flow block to reduce the injury.

Objective To accurately assess the condition of trauma patients at the emergency department (ED),the risk factors of in-hospital death were explored. Methods A total of 86 emergency trauma patients were retrospectively investigated. They were divided into survival and non-survival groups,in the First Hospital of China Medical University,from August 2016 to February 2017. Clinical parameters,such as sex,age,heart rate,oxygen saturation,mean arterial pressure,white blood cell count,serum creatinine,urea nitrogen, prothrombin time,activated partial thromboplastin time,hemoglobin,platelet count,serum albumin,fibrinogen,glutamic-pyruvic,total bilirubin,Glasgow coma scale (GCS),sequential organ failure assessment (SOFA) score,and injury severity score were evaluated and recorded. The parameters which were significantly different (P < 0. 1) between the two groups were analyzed using the logistic regression analysis to determine the independent risk factors of death at the ED. Receiver-operating characteristic curves were drawn to evaluate their prognostic abilities. Results GCS and SOFA score were the independent risk factors of in-hospital death in trauma patients (P < 0. 05). Conclusion Organ function,especially that of the brain,is closely related to the prognosis of adult trauma patients.