AIM:To investigate the effects of the Qingfei-Jiedu-Huatan formula(QJHF)on damage to the lung vascular endothelial barrier induced by Klebsiella pneumoniae in mice with severe pneumonia,as well as to elucidate its underlying mechanisms.METHODS:Fifty-one C57BL/6J mice were randomly divided into control group(n=6),model group(n=15),QJHF group(n=15),and ceftriaxone sodium(CRO)group(n=15).Severe pneumonia was in-duced in the mice by a single tracheal intubation with 50 μL of 1×1011 CFU/mL Klebsiella pneumoniae on day 0.Six hours after modeling,the mice in QJHF and CRO groups received their respective treatments,while those in control and model groups were administered an equal volume of saline.All mice were sacrificed on day 3 after the end of gavage.Lung histo-pathological changes were assessed using hematoxylin-eosin(HE)staining.Levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6 in lung tissues were measured by enzyme-linked immunosorbent assay(ELISA).Flow cytometry was used to detect CD11b+Ly6g+cells in bronchoalveolar lavage fluid(BALF).Proteomics and network pharma-cology analyses were conducted to elucidate the mechanisms of drug action.Western blot was conducted to assess the ex-pression levels of vascular endothelial cadherin(VE-cadherin),zonula occludens-1(ZO-1),occludin,vascular endothe-lial growth factor(VEGF),P38 mitogen-activated protein kinase(P38),and phosphorylated P38(p-P38)in lung tis-sues.RESULTS:Treatment with QJHF significantly attenuated the symptoms such as mental status and respiratory dis-tress,reduced mortality,mitigated lung tissue lesions,and decreased levels of IL-6,TNF-α,IL-1β,as well as BALF to-tal protein concentration,total cell count and neutrophil content in a mouse model of severe pneumonia(P＜0.05 or P＜0.01).Additionally,QJHF increased the expression of VE-cadherin,ZO-1 and occludin proteins in lung tissues.Pro-teomic analysis demonstrated that QJHF modulated the expression of 129 proteins in the lung tissues of mice suffering from severe pneumonia.Network pharmacology identified 328 potential targets associated with 14 major bioactive components of QJHF and 1 665 genes related to severe pneumonia,with 125 overlapping genes between the two datasets.The construc-tion of a protein-protein interaction(PPI)network,along with Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of the regulated proteins and overlapping genes,indicated that QJHF primarily in-fluenced the PI3K-Akt,MAPK and Rap1 signaling pathways,as well as VEGFR.Western blot analysis showed that QJHF significantly inhibited the expression of VEGF and P38 in lung tissues(P＜0.05 or P＜0.01).CONCLUSION:Treatment with QJHF attenuates severe pneumonia in mice,potentially by inhibiting VEGF/P38 signaling to protect the vascular endothelial barrier.

Moloney leukemia virus 10(MOV10)gene knockout(MKO)mouse neuro 2a(N2a)cell lines was constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)gene editing technology.First,a recombinant plasmid pMD18T-U6-sgRNA expressing MOV10 gene-specific guide RNA(sgRNA)was constructed,and then pMD18T-U6-sgRNA and pMJ920-Cas9-eGFP were co-transfected into N2a.The results showed that the MKO N2a cell line had normal cell activity and cell proliferation ability.The infection test of the MKO N2a cell line was carried out using the rabies virus(RABV)and vesicular stomatitis virus(VSV)of the Rhabdoviridae family.The results showed that the replication level of the Rhabdoviridae virus in the MKO N2a cell line was significantly enhanced.The results showed that a MKO N2a cell line was successfully constructed in this study,which provided a preliminary basis for the exploration of the biological function and antiviral mechanism of MOV10 and the develop-ment of a recombinant viral vector vaccine with RABV/VSV as the vector.

Moloney leukemia virus 10(MOV10)gene knockout(MKO)mouse neuro 2a(N2a)cell lines was constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)gene editing technology.First,a recombinant plasmid pMD18T-U6-sgRNA expressing MOV10 gene-specific guide RNA(sgRNA)was constructed,and then pMD18T-U6-sgRNA and pMJ920-Cas9-eGFP were co-transfected into N2a.The results showed that the MKO N2a cell line had normal cell activity and cell proliferation ability.The infection test of the MKO N2a cell line was carried out using the rabies virus(RABV)and vesicular stomatitis virus(VSV)of the Rhabdoviridae family.The results showed that the replication level of the Rhabdoviridae virus in the MKO N2a cell line was significantly enhanced.The results showed that a MKO N2a cell line was successfully constructed in this study,which provided a preliminary basis for the exploration of the biological function and antiviral mechanism of MOV10 and the develop-ment of a recombinant viral vector vaccine with RABV/VSV as the vector.

AIM:To investigate the effects of the Qingfei-Jiedu-Huatan formula(QJHF)on damage to the lung vascular endothelial barrier induced by Klebsiella pneumoniae in mice with severe pneumonia,as well as to elucidate its underlying mechanisms.METHODS:Fifty-one C57BL/6J mice were randomly divided into control group(n=6),model group(n=15),QJHF group(n=15),and ceftriaxone sodium(CRO)group(n=15).Severe pneumonia was in-duced in the mice by a single tracheal intubation with 50 μL of 1×1011 CFU/mL Klebsiella pneumoniae on day 0.Six hours after modeling,the mice in QJHF and CRO groups received their respective treatments,while those in control and model groups were administered an equal volume of saline.All mice were sacrificed on day 3 after the end of gavage.Lung histo-pathological changes were assessed using hematoxylin-eosin(HE)staining.Levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6 in lung tissues were measured by enzyme-linked immunosorbent assay(ELISA).Flow cytometry was used to detect CD11b+Ly6g+cells in bronchoalveolar lavage fluid(BALF).Proteomics and network pharma-cology analyses were conducted to elucidate the mechanisms of drug action.Western blot was conducted to assess the ex-pression levels of vascular endothelial cadherin(VE-cadherin),zonula occludens-1(ZO-1),occludin,vascular endothe-lial growth factor(VEGF),P38 mitogen-activated protein kinase(P38),and phosphorylated P38(p-P38)in lung tis-sues.RESULTS:Treatment with QJHF significantly attenuated the symptoms such as mental status and respiratory dis-tress,reduced mortality,mitigated lung tissue lesions,and decreased levels of IL-6,TNF-α,IL-1β,as well as BALF to-tal protein concentration,total cell count and neutrophil content in a mouse model of severe pneumonia(P＜0.05 or P＜0.01).Additionally,QJHF increased the expression of VE-cadherin,ZO-1 and occludin proteins in lung tissues.Pro-teomic analysis demonstrated that QJHF modulated the expression of 129 proteins in the lung tissues of mice suffering from severe pneumonia.Network pharmacology identified 328 potential targets associated with 14 major bioactive components of QJHF and 1 665 genes related to severe pneumonia,with 125 overlapping genes between the two datasets.The construc-tion of a protein-protein interaction(PPI)network,along with Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of the regulated proteins and overlapping genes,indicated that QJHF primarily in-fluenced the PI3K-Akt,MAPK and Rap1 signaling pathways,as well as VEGFR.Western blot analysis showed that QJHF significantly inhibited the expression of VEGF and P38 in lung tissues(P＜0.05 or P＜0.01).CONCLUSION:Treatment with QJHF attenuates severe pneumonia in mice,potentially by inhibiting VEGF/P38 signaling to protect the vascular endothelial barrier.

To establish a simple,convenient,sensitive,and specific method for rapid detection of Zi-ka virus(ZIKV),the whole genome sequences of ZIKV isolated from different times and regions were analyzed.The specific primers and probes were designed based on the screened target se-quences located in the conserved region of the ZIKV NS5 gene.By combining RT-LAMP isother-mal amplification technology and immunochromatography technology,a reverse transcription loop mediated isothermal amplification nucleic acid and flow visualization strip(RT-LAMP-VF)detec-tion method for ZIKV was established.The results showed that the method had good specificity and sensitivity.When the ratio of inner,outer,and ring primers(FIP∶LF∶F3)was 4∶2∶1,the detection method can specifically detect 102 copies/pL RNA transcripts or 2.15 pfu ZIKV at 61 ℃for 45 minutes,with no cross reaction with other flaviviruses such as Japanese encephalitis virus and classical swine fever virus.Other RNAs in blood tissue samples did not affect the sensitivity and specificity of RT-LAMP-VF,indicating that the method can be applied to clinical practice.The ZIKV RT-LAMP-VF detection method established in this study is easy to perform and does not require special instruments and equipment.It is particularly suitable for the rapid detection of ZIKV in grassroots units,providing technical support and material support for the establishment of on-site rapid detection and early warning and prediction systems for ZIKV disease.

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.

Objective: To study the clinical application of three-dimensional (3D) visualization technology in liver resection of complicated liver cancer. Methods: A retrospective analysis of 28 patients with complicated liver cancer treated from June 2017 to June 2018 in the Department of Hepatobiliary Surgery, the Affiliated Tumor Hospital of Guangxi Medical University. There were 26 males and 2 females, aged (46±10) years old. A treatment plan on how to perform liver resection for these patients was developed under the guidance of 3D visualization technology. The actual surgical procedures, operation time, intraoperative blood loss, and postoperative complications were documented. The virtual resected liver volume was compared with the actual resected liver volume. The virtual surgical resection margin was also compared with the actual surgical resection margin. Results: All the 28 patients with complicated liver cancer completed the 3D visualization analysis with the location, shape and quantity of tumor being clearly shown. Of the 27 patients who underwent liver resection, 13 underwent anatomical hepatectomy, and 14 underwent non-anatomical hepatectomy. The operation time ranged from 145 to 350 min (median 240 min). The intraoperative blood loss ranged from 100 to 1 500 ml (median 300 ml). The incisional wound healed slowly in 4 patients, pleural effusion developed in 8 patients, and ascites in 2 patients. There were no significant differences in the virtual resected liver volume compared with the actual resected liver volume (P>0.05). There was an excellent positive correlation between the patient's virtual resected liver volume and the actual resected liver volume (r=0.986, P<0.05). There was no significant difference between the virtual surgical resection margin and the actual surgical resection margin (P>0.05). There was an excellent positive correlation between virtual surgical resection margin and the actual surgical resection margin (r=0.983, P<0.05). Conclusion Three-dimensional visualization technology accurately assessed the liver status, optimized surgical procedures, and played an important role in liver resection of complex liver cancer.

Objective To compare the differences between electronic lesion of pyramidal tract and internal capsule hemorrhage in establishing spastic rat models.Methods Experimental animals were randomly assigned into three groups:electronic lesion group (EL),internal capsule hemorrhage group (ICH) and control group (CON,n=28).Seven,14,21 and 28 d after each treatment,the general conditions of the rats and behavioral tests on them were recorded; and the gastrocnemius Hoffman reflex (H reflex) was detected; and the percentage of type Ⅰ muscle fiber in quadriceps femoris were examined by skeletal muscle myosin ATP staining.Results Rats in EL group and ICH group showed significant differences in behavioral tests as compared with CON group (P＜0.05); the duration of symptoms of neurological deficit was significantly longer in EL group than the other two groups (P＜0.05).In contrast to CON group,the amplitude of gastrocnemius H reflex in EL group and ICH group was increased and the latency of H reflex was decreased with significant difference (P＜0.05),which could be observed till 28 d and 14 d,respectively,after each treatment.The percentage of type Ⅰ muscle fiber in quadriceps femoris EL and ICH groups was obviously increased as compared with that in CON group (P＜0.05),which persisted to 21 and 14 d,respectively,after each treatment.Meanwhile,rats in EL group showed significant differences in behavioral test,gastrocnemius H reflex and percentage of type] muscle fiber in quadriceps femoris as compared with ICH rats (P＜0.05).Conclusion The muscle tone of EL rats is higher,and lasts longer than that ofICH rats.