Objective To construct a prostate cancer specific oncolytic adenovirus armed with a fusion protein gene , PSA-IZ-CD40L, and to evaluate its oncolytic efficiency and immune activation ability in vitro.Methods Prostate Specific Antigen (PSA) gene, CD40L-N and CD40L-C genes were obtained from cDNA of LNCaP cells and Jurkat cells using poly-merase chain reaction (PCR) or nested-PCR, respectively.PSA,Linker,CD40L-N and CD40L-C were linked sequentially to generate fusion protein gene PSA-IZ-CD40L (PL) by overlapping PCR.Then, prostate specific oncolytic adenovirus PL-carrying gene, Ad-PL-PPT-E1A,was constructed using the oncolytic adenovirus system , which was based on Adeasy sys-tem.PC3M cells were infected by Ad-PL-PPT-E1A at serial multiplicity of infection (MOI), and the apoptosis was detec-ted by flow cytometry at several time points post-infection.For immune activation detection , PC3M cells were infected with Ad-PL-PPT-E1A at a MOI of 50, and the cell lysate was collected at 48 h post-infection.Peripheral blood mononuclear cells derived (PBMCs) from healthy donors were stimulated by the lysate from PC 3M cells or Ad-PL-PPT-E1A infected PC3M cells before proliferation was assayed using cell counting kit-8 (CCK8).Results Fusion protein gene, PSA-IZ-CD40L, was successfully constructed and cloned into the prostate cancer specific adenovirus to generate Ad -PL-PPT-PL. The expression of E1A and PL protein could be detected by reverse transcription PCR and Western-blotting.Cytopathic effect was observed in PC3M cells infected with Ad-PL-PPT-E1A.Furthermore, the apoptosis rate reached 70.67% ± 2.98%at 48 h post-infection with 200 MOI Ad-PL-PPT-E1A.Compared with the lysate of PC3M cells, that from Ad-PL-PPT-E1A infected cells could promote the proliferation of PBMCs .Conclusion We have constructed a prostate cancer spe-cific oncolytic adenovirus armed can fusion protein gene PL , Ad-PL-PPT-E1A, which could kill PC3M cells effectively and enhance the proliferation of PBMCs in vitro.

Objective: To explore the prevalence and associated distress of lower urinary tract symptoms among female college students in Xuzhou. Methods: Convenience sampling was used to recruit 1 000 female college students in Xuzhou China. Chinese versions of International Consultation on Incontinence Questionnaire LUTS and General Information Questionnaire was used. Results: The prevalence of lower urinary tract symptoms was 95.7%. The top ten prevalence rates were urgency (88.2%), hesitancy (65.4%), nocturia (48.5%), intermittent stream (47.3%), straining (30.5%), increased daytime frequency (30.1%), bladder pain (21.0%), SUI (18.5%), UUI (13.9%), nocturnal enuresis (7.5%). Urinary urgency was the most disturbing symptom (49.0%). Risk factors of urinary urgency included rural household registration (OR=1.67, 95%CI:1.10-2.55, P=0.017) and premature urination (OR=1.65, 95%CI:1.23-2.21, P=0.00). Conclusion Female college students have a high prevalence of lower urinary tract symptoms but with moderate severity and distress. Toileting behaviors affect bladder health, which require early intervention.

Objective:To observe the effect of gait training assisted by an end-effector-based robot on the gait, balance, walking and general lower limb motor functioning of stroke survivors.Methods:Thirty-nine stroke survivors were randomized into a training group ( n=21) and a control group ( n=18). In addition to routine hemiplegia rehabilitation, the control group was given medium frequency electrical stimulation while the training group underwent 20 minutes of gait training assisted by an end-effector-based robot six days a week for 4 weeks. Before and after the intervention, both groups were evaluated using the functional ambulatory categories (FAC), the Fugl-Meyer lower extremity assessment, the Wisconsin Gait Scale and the Berg balance scale (BBS). Results:There were no significant differences between the groups before the intervention. Afterward significant improvement was observed in both groups in all measurements, with the training group scored better in terms of FAC and BBS.Conclusion:Using an end-effector-based robot in gait training might be more advantageous for improving the gait, balance, walking and general lower limb functioning of stroke survivors.

Objective:To explore the molecular basis for an individual with Bel subtype of the ABO blood type due to a novel c. 620T>C variant gene, and assess its impact on the structure of GTB transferase.Methods:An individual who had visited the First Affiliated Hospital of Zhengzhou University on February 11, 2023 was selected as the study subject. ABO phenotyping was initially conducted with serological methods, which was followed by direct sequencing of 7 exons of the ABO gene. Subsequently, single-strand sequencing was carried out by using allele-specific primers, and the variant in the B transferase was homology-modeled using the Modeller software. The impact of the variant on the transferase′s spatial structure was analyzed with the PyMOL software. Results:The serological phenotype of the patient was identified as the Bel subtype. Direct sequencing revealed that she has harbored a novel c. 620T>C variant, resulting in a p. Leu207Pro substitution in the polypeptide chain. Combined with single-strand sequencing, her genotype was ultimately determined as ABO* BELnew/ ABO* O.01.02. Three-dimensional protein structure modeling showed that, compared with the wild type, the distance of one hydrogen bond between Proline and Glycine at position 272 has increased, along with disappearance of another hydrogen bond. Conclusion:The novel c. 620T>C (p.Leu207Pro) variant of B allele may affect the structural stability of the glycosyltransferase. The weakened enzyme activity in turn may lead to reduced B antigen expression, manifesting as the Bel subtype by serological analysis.

Objective:To study the molecular basis for a proband with A subtype B of the ABO blood group and explore the influence of amino acid variant on the activity of glycosyltransferase (GT).Methods:A proband who had presented at the First Affiliated Hospital of Zhengzhou University on July 2, 2020 was selected as the study subject. Serological identification of the ABO blood groups of the proband and her family members were performed by gel card and test tube methods. The ABO gene of the proband was identified by PCR-sequence specific primers (PCR-SSP) and DNA sequencing. A 3D molecular homologous model was constructed to predict the impact of the variant on the stability of α-(1→3)-D-N-acetylgalactosamine transferase (GTA). Results:The red blood cells of the proband, her mother and two younger brothers showed weak agglutination with anti-A and strong agglutination with anti-B. The sera showed 1~2+ agglutination with Ac and no agglutination with Bc. Based on the serological characteristics, the proband was identified as AwB subtype. Pedigree analysis suggested that the variant was inherited from her mother. The blood group of the proband was identified as A223B type by PCR-SSP. ABO gene sequencing analysis showed that the proband has harbored heterozygous variants of c. 297A>G, c. 467C>T, c. 526C>G, c. 657C>T, c. 703G>A, c. 796C>A, c. 803G>C, c. 930G>A and c. 1055insA. Based on the results of clone sequencing, it was speculated that the genotype was ABO* A223/ ABO* B.01. There were c. 467C>T and c. 1055insA variants compared with ABO* A1.01, and c. 1055insA variant compared with ABO* A1.02. Homologous modeling showed that the C-terminal of A223 GT was significantly prolonged, and the local amino acids and hydrogen bond network have changed. Conclusion:Above results revealed the molecular genetics mechanism of A223B subtype. The c. 1055insA variant carried by the proband may affect the enzymatic activity of GTA and ultimately lead to weakening of A antigen.

Objective To understand the management status of emergency and life support medical equipment in town-ship health centers in Huazhou City,providing a reference for improving emergency service levels in grassroots medical institu-tions.Methods A convenience sampling method was used to conduct a questionnaire survey of physicians,nurses,medical technicians,public health personnel,and administrative and logistical staff in 24 township health centers in Huazhou City.The survey focused on seven aspects:procurement and configuration,upgrades,operational use,maintenance,emergency handling,management systems,and quality assessment and monitoring.Results Training on emergency and life support equipment was generally adequate in Huazhou City(65.2％).Health centers in northern and central Huazhou performed better than those in the urban and southern areas regarding equipment training,emergency handling procedures for equipment failures,regular emergency drills,meeting clinical treatment needs,management system completeness,routine maintenance and inspections,and regular quality assessments.Factor analysis revealed significant issues with equipment aging and quality in urban township health cen-ters,while northern township health centers urgently needed life-sustaining and therapeutic equipment.Conclusion There are regional disparities in the management levels of emergency and life support medical equipment in township health centers in Hua-zhou City.It is recommended to enhance the overall quality of personnel in township health centers,optimize"effective training"for staff,and improve annual management plans to further elevate grassroots emergency service capabilities.

Gene editing technology is a genetic operation technology that can modify the DNA sequence at the genomic level. The precision gene editing technology based on CRISPR/Cas9 system is a gene editing technology that is easy to operate and widely used. Unlike the traditional CRISPR/Cas9 system, the precision gene editing technology can perform site-directed mutation of genes without DNA template. This review summarizes the recent development of precision gene editing technology based on CRISPR/Cas9, and prospects the challenges and opportunities of this technology.
Gene Editing ; CRISPR-Cas Systems/genetics* ; Mutation ; Genome

Objective: To propose the concept of a novel regional control and prevention (RCP) system for the outbreak of COVID-19 infectious disease, design an emergency epidemic prevention information system based on the existing network architecture and information system in the region, and a remote intelligent medical consultation and remote office platform, research and develop the technology of risk assessment and early warning for people in the region, and improve the regions’prevention and control ability facing emergency of major infectious diseases. Methods: Taking colleges, affiliated (teaching) hospitals, and cloud applications as typical RCP regional units, the existing local area network interaction methods between the cloud and universities and affiliated (teaching) hospitals are established to realize remote work in the network environment, remote medical imaging, psychological and ethical consultation and interaction; applying multi-agent propagation model based on complex network, combining Global Positioning System (GPS), Radio Frequency Identification (RFID), and electronic fence technology, to realize the risk classification and early warning of units and personnel in the area. Results: In the RCP, a system architecture combining campus network, affiliated (teaching) hospital intranet, and the Internet is used. Dynamic connection is made using distributed technology and cloud storage. The data buffer mechanism of the intermediary database in the network realized telemedicine consultation and telecommuting. Relying on the platform, multi-agent propagation model based on complex network and cellular automaton model are used to realize the score and early warning of population exposure risk in the region by using GPS, RFID and electronic fence technology. Conclusions In the epidemic phase of major infectious diseases, the construction of RCP can improve the response speed of wartime epidemic prevention, provide reasonable data-based warnings and risk ratings, and reduce the exposure risk of susceptible people. The design and development of RCP is a systematic project that needs to combine regional structural and functional characteristics, and the foundation of the early informatization work in the region and the level of the emergency development team determine the development progress, maintenance, and actual application effects. It is recommended to establish a peacetime and wartime combined RCP mode and incorporate it into the government's disease control system to improve the national and regional level of prevention and control of major infectious diseases.