[Objective]To detect the sensitivity of multidrug-resistant Acinetobacter baumannii(MDRAB)to berberine and investigate the effect of berberine on its resistance.[Methods]Sixty clinical strains of MDRAB obtained from Microbiology Laboratory of Zhejiang Provincial Hospital of Traditional Chinese Medicine between January and December 2023 were utilized.Microbroth dilution was used to measure the minimum inhibitory concentration(MIC)of berberine against MDRAB.Protein profiles of both the original and passaged strains were examined through matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),carbapenemase related genes were detected simultaneously.[Results]The MIC values of 60 MDRAB strains to berberine were all more than or equal to 5 mg·mL-1.After the action of berberine,MALDI-TOF MS analysis of the two groups of strains revealed that the peaks at mass-to-charge ratios of 6 644.47,12 297.67,and 16 985.72 were characteristic peaks with significant differences between the two groups.The detection rate of carbapenemase imipenemase metallo-β-lactamase(IMP)genes decreased from 34.2％to 10.5％,with a statistically significant difference(P＜0.05).[Conclusion]When used alone,berberine has very limited ability to directly inhibit or kill MDRAB;berberine can reverse the drug resistance mechanism of MDRAB by altering the expression of MDRAB proteins and IMP genes of carbapenemases,providing a new approach for the clinical treatment of MDRAB infections.

At present, robotic surgery system, as a cutting-edge technique in minimally invasive surgery, has become a crucial component in the training programs for outstanding surgeons. Given the relatively late start of this technique and immature faculty resources, there is an urgent need to optimize and improve the teaching philosophies and methodologies for robotic skill training in China, and Sino-foreign cooperation has emerged as a significant pathway to promote this reform. Taking the "International Robotic Surgery Training Center (IRSTC)" jointly established by The First Affiliated Hospital of Sun Yat-sen University and University of California as an example, this article provides a comprehensive analysis of collaboration background, construction of the training system, curriculum design, cooperation channels, and localization practices, in order to provide a reference and valuable experiences for Sino-foreign cooperation in clinical skill training.

Objective:To obtain a prevalent respiratory syncytial virus (RSV) clinical isolate in Beijing and analyze the genotype and biological characteristics of the strain.Methods:A nasopharyngeal secretion specimen was collected from a child with RSV infection in Beijing in 2023 and used for viral isolation. Viral nucleic acid was amplified using qRT-PCR. The isolated virus was identified by transmission electron microscopy, indirect immunofluorescence assay, and plaque formation assay. A phylogenetic analysis was conducted based on the whole-genome sequencing results. Virus titers were determined, and replication characteristics were analyzed. The efficacy of the isolated strain for in vitro screening of antiviral drugs was validated. Results:A clinical RSV isolate, named hRSV/C-Tan/BJ 202301, was successfully isolated, which could form syncytia in Hep-2 cells. Spherical, filamentous, and irregular virus particles were observed by electron microscopy. Immunofluorescence detection showed green fluorescence in Hep-2 cells, and plaque assay showed round plaques, which were similar to the Long strain in morphology. Genomic sequence analysis showed that it belonged to ON1 genotype. It exhibited similar cell growth kinetics characteristics with the Long strain and could be used for antiviral drug screening in vitro. Conclusions:In this study, one RSV strain is successfully isolated and identified. The biological characteristics and the phylogenetic relationship of this strain reflect the characteristics of the circulating strains in Beijing, which provides experimental material for RSV vaccine development and antiviral drug screening in China.

Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.

Terahertz(THz)technology,as an emerging tool for characterizing biomacromolecules,has rapidly developed over the past few decades,demonstrating significant application potential in multiple fields.Defensive qi is a vital energy in the human body,responsible for defense functions by warming the skin,regulating the opening and closing of the striae and interstices,and resisting the invasion of external pathogens.Defensive qi is one of the core concepts for disease prevention and health maintenance in traditional Chinese medicine(TCM).However,due to the high abstraction and complexity of TCM theory,the scientific connotation and mechanism of action of defensive qi remained incompletely elucidated.THz waves have the unique physical properties of molecular vibration sensitivity,penetrability,and thermal radiation effects,and are highly sensitive to biological tissues,which make THz technology a promising tool for defensive qi research.By analyzing the characteristics of THz waves,this paper explores the feasibility of applying THz technology to investigate the circulation patterns of defensive qi,the prevention and treatment of disorders related defensive qi,the pathological essence of defensive qi diseases,the material basis of defensive qi,the screening of defensive qi disorders,and the development of novel drugs targeting defensive qi disorders.This approach provides a new perspective and methodology for the research of TCM fundamental theory.

Human alphaherpesvirus 2 (HSV-2) is the main pathogen resulting human genital herpes, which poses a major threat to the socio-economic development, while there is no effective vaccine. In this study, we developed a novel lipopolyplex (LPP)-delivered mRNA vaccine expressing the HSV-2 envelope glycoprotein gD and evaluated its immunogenicity in mice. The mRNA vaccine was prepared from the genetically modified gD mRNA synthesized in vitro combined with the LPP delivery platform and it was named gD-ORI mRNA. The expression of gD antigen in the mRNA vaccine was validated in vitro by Western blotting and indirect immunofluorescence assay, then the immune responses induced by this mRNA vaccine in mice were evaluated. The immunization with gD mRNA alone induced strong humoral and cellular immune responses in mice. Robust and long-lasting gD-specific IgG antibodies were detected in the mouse serum after booster immunization with gD-ORI mRNA. The immunized mice exhibited a Th1/Th2 balanced IgG response and robust neutralizing antibodies against HSV-2, and a clear dose-response relationship was observed. The gD-specific IgG antibodies were maintained in mice for a long time, up to 18 weeks post-booster immunization. At the same time, multifunctional gD-specific CD4⁺ and CD8⁺ T cells in vaccinated mice were detected by intracellular cytokine staining (ICS). This novel gD-expressing mRNA vaccine delivered by LPP induces strong and long-lasting immune responses in mice post booster immunization and has a promising prospect for development and application. This study provides scientific evidence and reference for the development of a new mRNA vaccine for HSV-2.
Animals ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Mice ; Herpes Genitalis/immunology* ; RNA, Messenger/immunology* ; Female ; Mice, Inbred BALB C ; Antibodies, Viral/blood* ; mRNA Vaccines/immunology* ; Antibodies, Neutralizing/blood* ; Humans

Objective To compare the clinical efficacy and safety of lumbar intervertebral space implantation with single and two fusion devices in treatment of lumbar instability under intervertebral foraminoscopy.Methods 101 patients(101 vertebral bodies)diagnosed with L4,L5 or S1 segment Ⅰ° to Ⅱ° spondylolisthesis from January 2019 to June 2022 were enrolled and the patients were randomly divided into group A(single fusion device group)and group B(double fusion devices group)by intervertebral foraminoscopy.Among of them,there were 51 cases of vertebral spondylolisthesis in group A and 50 cases of vertebral spondylolisthesis in group B.The patients were followed up at 1 week,1 month,6 months,9 months and 15 months after surgery,and the differences in spondylolisthesis distance,intervertebral space height,intervertebral space fusion,vertebral sedimentation rate,lumbar pain visual analogue scale(VAS)and lumbar Oswerry disability index(ODI)between the two groups were compared.Results All the 101 patients were followed up throughout the whole process,and there were no significant differences in operation time,intraoperative blood loss,and postoperative drainage between the two groups(P＞0.05),and the VAS was(7.32±0.57)in group A and(7.14±0.61)in group B one week after operation,there was no significant difference between the two groups(P＜0.05).At 6 months after surgery,the ODI of group A was(27.44±9.52)%and group B was(21.89±8.46)%,and there was no significant difference between the two groups(P＞0.05).And at 6,9 and 15 months follow-up,the fusion rates of group A were 11.76%,68.63%and 90.20%,respectively,and the fusion rates of group B were 18.00%,80.00%and 96.00%,respectively,and there was significant difference between the two groups(P＜0.05).At 15 months postoperative follow-up,the intervertebral space height of group A(0.45±0.07)cm was significantly smaller than that of group B(0.58±0.06)cm,and the difference was statistically significant(P=0.038).There was no significant difference between group A and group B(0.33±0.18)mm and(0.28±0.21)mm(P=0.079)of spondylolisthesis distance.Conclusion Implantation of the double fusion devices can provide more contact area and better stability,shorten the vertebral fusion time,increase the vertebral fusion rate,and reduce the occurrence of vertebral settlement.It is worthy of clinical promotion and application.

At present, robotic surgery system, as a cutting-edge technique in minimally invasive surgery, has become a crucial component in the training programs for outstanding surgeons. Given the relatively late start of this technique and immature faculty resources, there is an urgent need to optimize and improve the teaching philosophies and methodologies for robotic skill training in China, and Sino-foreign cooperation has emerged as a significant pathway to promote this reform. Taking the "International Robotic Surgery Training Center (IRSTC)" jointly established by The First Affiliated Hospital of Sun Yat-sen University and University of California as an example, this article provides a comprehensive analysis of collaboration background, construction of the training system, curriculum design, cooperation channels, and localization practices, in order to provide a reference and valuable experiences for Sino-foreign cooperation in clinical skill training.

[Objective]To detect the sensitivity of multidrug-resistant Acinetobacter baumannii(MDRAB)to berberine and investigate the effect of berberine on its resistance.[Methods]Sixty clinical strains of MDRAB obtained from Microbiology Laboratory of Zhejiang Provincial Hospital of Traditional Chinese Medicine between January and December 2023 were utilized.Microbroth dilution was used to measure the minimum inhibitory concentration(MIC)of berberine against MDRAB.Protein profiles of both the original and passaged strains were examined through matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),carbapenemase related genes were detected simultaneously.[Results]The MIC values of 60 MDRAB strains to berberine were all more than or equal to 5 mg·mL-1.After the action of berberine,MALDI-TOF MS analysis of the two groups of strains revealed that the peaks at mass-to-charge ratios of 6 644.47,12 297.67,and 16 985.72 were characteristic peaks with significant differences between the two groups.The detection rate of carbapenemase imipenemase metallo-β-lactamase(IMP)genes decreased from 34.2％to 10.5％,with a statistically significant difference(P＜0.05).[Conclusion]When used alone,berberine has very limited ability to directly inhibit or kill MDRAB;berberine can reverse the drug resistance mechanism of MDRAB by altering the expression of MDRAB proteins and IMP genes of carbapenemases,providing a new approach for the clinical treatment of MDRAB infections.

Objective To compare the clinical efficacy and safety of lumbar intervertebral space implantation with single and two fusion devices in treatment of lumbar instability under intervertebral foraminoscopy.Methods 101 patients(101 vertebral bodies)diagnosed with L4,L5 or S1 segment Ⅰ° to Ⅱ° spondylolisthesis from January 2019 to June 2022 were enrolled and the patients were randomly divided into group A(single fusion device group)and group B(double fusion devices group)by intervertebral foraminoscopy.Among of them,there were 51 cases of vertebral spondylolisthesis in group A and 50 cases of vertebral spondylolisthesis in group B.The patients were followed up at 1 week,1 month,6 months,9 months and 15 months after surgery,and the differences in spondylolisthesis distance,intervertebral space height,intervertebral space fusion,vertebral sedimentation rate,lumbar pain visual analogue scale(VAS)and lumbar Oswerry disability index(ODI)between the two groups were compared.Results All the 101 patients were followed up throughout the whole process,and there were no significant differences in operation time,intraoperative blood loss,and postoperative drainage between the two groups(P＞0.05),and the VAS was(7.32±0.57)in group A and(7.14±0.61)in group B one week after operation,there was no significant difference between the two groups(P＜0.05).At 6 months after surgery,the ODI of group A was(27.44±9.52)%and group B was(21.89±8.46)%,and there was no significant difference between the two groups(P＞0.05).And at 6,9 and 15 months follow-up,the fusion rates of group A were 11.76%,68.63%and 90.20%,respectively,and the fusion rates of group B were 18.00%,80.00%and 96.00%,respectively,and there was significant difference between the two groups(P＜0.05).At 15 months postoperative follow-up,the intervertebral space height of group A(0.45±0.07)cm was significantly smaller than that of group B(0.58±0.06)cm,and the difference was statistically significant(P=0.038).There was no significant difference between group A and group B(0.33±0.18)mm and(0.28±0.21)mm(P=0.079)of spondylolisthesis distance.Conclusion Implantation of the double fusion devices can provide more contact area and better stability,shorten the vertebral fusion time,increase the vertebral fusion rate,and reduce the occurrence of vertebral settlement.It is worthy of clinical promotion and application.