Objective To express the capsid(Cap) protein of porcine circovirus 2(PCV2) by insect-baculovirus expression system, evaluate its immunogenicity as a vaccine candidate, and explore whether it can induce humoral and cellular immune responses in BALB/c mice.Methods Following codon optimization, the PCV2 Cap gene was cloned into pFastBac Dual vector and transformed into E.coli DH10 Bac cells. After Bacmid extraction, PCR amplification was carried out with M13 universal primers. The appropriately identified Bacmid was then transfected into Sf9 cells to express Cap protein, and the protein was quantified. Forty-five female BALB/c mice were randomly divided into three groups: PBS control, Cap and commercial vaccine YI Cap group, 15 mice in each group, which were immunized subcutaneously on the back once at 14-day intervals, for a total of three times, with PBS 100 ??L, Cap 10 ??g, and commercial vaccine YI Cap 10 ??g respectively each time. The serum IgG level of mice was measured by ELISA, the cellular immune response type was analyzed by flow cytometry, and the histocompatibility of the subunit vaccine was observed by H&E staining.Results The insect-baculovirus expression system effectively expressed the target protein, and the Cap protein with enhanced green fluorescent protein(eGFP) colocalized with the cell's plasma membrane. Transmission electron microscopy revealed that the Cap protein could self-assemble to produce virus-like particles(VLPs) before constructing another recombinant vector sans the eGFP fluorescent flag. SDS-PAGE and Western blot analysis revealed the conspicuous bands at the desired area. The IgG content of Cap vaccine candidate mice reached the highest level at 42 days. The levels of IFNγ and IL-4 were(27. 75 ± 0. 99) and(17. 63 ± 1. 13) pg/mL, respectively, both of which were significantly higher than those of PBS group(F = 14. 89 and 0. 211 9,P < 0. 05 and < 0. 001, respectively). The stimulation index(SI) of splenic lymphocytes in the Cap group reached 6. 72, which was significantly higher than that in the PBS group(F = 1. 228, P < 0. 001). The Cap vaccine candidate could induce Th1-type cellular immune response and exhibited good histocompatibility.Conclusion The Cap protein was successfully expressed in the insect-baculovirus expression system. The Cap vaccine candidate can induce humoral and cellular immune responses in mice after immunization, which is a potential vaccine candidate for further development of more effective vaccines against PCV-related diseases.

Objective To investigate the perioperative safety of patients with myasthenia gravis who take high doses of oral glucocorticoids. Methods A retrospective analysis was conducted on the clinical data of patients with myasthenia gravis who received oral glucocorticoids and underwent thoracoscopic thymectomy at the Department of Thoracic Surgery, the University of Hong Kong-Shenzhen Hospital from April 2013 to October 2019. Patients were divided into a high-dose steroid group and a medium-to-low dose steroid group based on the dosage of oral steroids, and the clinical data of the two groups were compared. Results A total of 102 patients were included, including 19 (18.62%) males and 83 (81.37%) females, with an average age of (32.25±9.83) years. There were 75 patients in the medium-to-low dose steroid group and 27 patients in the high-dose steroid group. All patients in both groups successfully completed the surgery without major intraoperative bleeding, conversion to open chest surgery, delayed extubation, severe infection, or perioperative death. The daily oral steroid dose for the high-dose steroid group was (35.81±4.29) mg, and for the medium-to-low dose steroid group it was (15.29±2.17) mg. There was no statistical difference in the operation time [(124.69±23.51) min vs. (117.89±21.46) min, P=0.172] and intraoperative blood loss [(21.19±3.48) mL vs. (20.56±3.41) mL, P=0.419] between the two groups. Postoperatively, 12 (11.76%) patients developed complications: one patient of myasthenic crisis (the medium-to-low dose steroid group), which was improved after short-term respiratory support and intravenous immunoglobulin treatment; 11 patients of respiratory/swallowing difficulties (9 in the medium-to-low dose steroid group and 2 in the high-dose steroid group), which were improved after anticholinergic treatment to reduce oral secretions and sputum suction, and the patients were discharged smoothly. There was no statistical difference in the incidence of postoperative complications between the two groups (P=0.637). Conclusion On the basis of good perioperative management, it is safe and feasible for patients with myasthenia gravis who take high dose of oral steroids to undergo thymectomy, and they have the same perioperative safety as patients with medium-to-low dose steroids.

ObjectiveTo compare the differences between the ultrafiltration method and the dextran gel filtration method during the purification of Tfn-modified PCL micelles by using purification efficiency and micelle purity as indicators. MethodsCoumarin-6 （C6） was used as a fluorescent probe and was loaded into HOOC-PEG-PCL to form PCL micelles by the film-dispersion method. Tfn was then conjugated to the surface of PCL micelles via an amidation reaction， resulting in two types of micelles： Tfn/PCL_H and Tfn/PCL_L. The pharmaceutical properties of the two types of micelles were characterized. The micelles were then purified through ultrafiltration method and dextran gel method respectively， and the efficiency of the two methods， along with the purity of the final micelles， was compared. The density of Tfn on the surface of PCL micelles was also calculated. ResultsThe hydrated diameter of PCL micelles was approximately 73 nm， and the C6 loading efficiency was around 0.046%. The size increased to 134 nm and 158 nm for Tfn/PCL_L and Tfn/PCL_H， respectively. The micelle population was monodisperse. The purification results showed that， for the ultrafiltration method， after two and one rounds of purification， the Tfn/C6 ratio stabilized at 23.6 and 3.4 for Tfn/PCL_H and Tfn/PCL_L， respectively. For the dextran gel filtration method， the Tfn/C6 ratio reached 23.7 for the Tfn/PCL_H group after two rounds of purification. However， for the Tfn/PCL_L group， the Tfn/C6 ratio increased during four rounds of dextran gel purification， and a significant difference （P = 0.042 4） was observed between the first and last filtrations. The density of Tfn in the final micelles were calculated. For the ultrafiltration method， the Tfn density of Tfn/PCL_H and Tfn/PCL_L were 94.9% and 13.8%， respectively. For the dextran gel filtration method， the density of the two micelles were 95.6% and 14.4%， respectively. For Tfn/PCL_L group， the density results revealing a statistically significant difference （P=0.000 2）. ConclusionThe purification efficiency of the two methods is comparable. However， the purity of the final micelles shows a significant difference， with the dextran gel filtration method resulting in higher purity， particularly for the Tfn/PCL_L micelles.

Objective To investigate the occurrence and characteristics of Antineutrophil cytoplasmic antibody(ANCA)associated vasculitis induced by methimazole,and to provide references for clinical safe drug use.Methods Case reports of ANCA associated vasculitis induced by methimazole published in Wanfang,CNKI,PubMed,and Web of Science were searched from the inception to October 31 st,2023.Demographic characteristics,drug use,complications,treatment and outcome were analyzed using descriptive statistical method.Results A total of 14 patients from 14literature were included.There were 3 males and 11 females with ages ranging from 8 to 79 years,with a mean age of(47.79±23.47)years.Four patients developed symptoms within 1 year,nine patients developed symptoms from 2 to 12 years,and 1 patient developed symptoms 24 years after medication.ANCA associated vasculitis affected kidney in 5 patients,lung and skin in 5patients,vision in 2 patients and heart in 2 patients.All patients discontinued methimazole,2 patients improved spontaneously without treatment,1 patient improved after anti-infection,and all others received hormonal or immunosuppressive therapy.1 patient developed death,and all others improved or were cured after treatment.Conclusions ANCA-associated vasculitis is a rare adverse reactionof methimazole.Most patients have a long latency period before the onset of disease,mainly involving multiple organs such asskin,kidney,lung,and eyesight.Clinicians should pay attention to differentiate it from primary vasculitis and discontinue the drug as soon as possible.When serious organ damage occurs,glucocorticoids and immunosuppressants should be adminstered promptly to avoid aggravation of the disease and endangerment of life.

Objective:Based on the activities of Helicobacter pylori ( HP) screening in Jiangsu Province in 2024, to evaluate the overall management ability in HP screening, testing, treatment and follow-up in primary medical facilities. Methods:From May 15 to October 18, 2024, the data of HP screening and treatment were retrospectively collected from 79 township health centers, community hospitals, and community health service centers in Jiangsu Province. The rates of screening completion, urea breath test (UBT) completion, treatment rate, UBT follow-up completion, and HP eradication were analyzed. Chi-square test was used for statistical analysis. Results:The completion rate of HP screening was 94.45% (15 489/16 400). There were 6 604 cases (42.64%) with serum HP antibody positive among the 15 489 individuals who completed screening. The positive rate of serum HP antibody in males was higher than that in females (44.77%, 2 643/5 904 vs. 41.32%, 3 961/9 585), and the difference was statistically significant ( χ2=17.69, P<0.001). The positive rates of serum HP antibody in screened individuals aged 18 to 19, 20 to 39, 40 to 59, and 60 to 75 years old were 22.38% (32/143), 36.12% (1 168/3 234), 45.01% (3 240/7 199), and 44.05% (2 164/4 913), respectively, and the difference was statistically significant( χ2=100.73, P<0.001). Among the 6 604 HP antibody-positive individuals, 4 381 cases completed UBT, with a UBT completion rate of 66.34% (4 381/6 604). There were 3 197 individuals with both HP serum antibody and UBT positive, the consistency rate of the 2 tests was 72.97% (3 197/4 381). Totally 2 737 cases received treatment, with a treatment completion rate of 85.61% (2 737/3 197); 2 327 individuals underwent UBT follow-up, with a follow-up completion rate of 85.02% (2 327/2 737). During follow-up, the result of UBT was negative in 1 982 individuals, and the HP eradication rate was 85.17% (1 982/2 327). Conclusions:There are deficiencies in the completion rate of HP screening, testing, treatment, and follow-up in primary hospitals, especially in the completion rate of UBT, which may be related to cognitive insufficiency for HP in residents. It is necessary to strengthen the training of physicians′ abilities in primary hospitals, optimize the allocation of drug resources, enhance health education, and increase residents′ participation and compliance.

Objective:To investigate the clinical application effect and preliminary experience of ureteroscopy-assisted flexible ureteroscopic lithotripsy in the treatment of transplanted kidney stones.Method:A retrospective analysis was conducted on the clinical data of 9 kidney transplant recipients with graft stones who underwent ureteroscopy-assisted flexible ureteroscopic lithotripsy at the First Affiliated Hospital of Zhengzhou University between January 2020 and January 2023. The recipients' general information, surgical procedures, and postoperative outcomes were reviewed and summarized. Additionally, a comprehensive literature search was performed in both Chinese and international databases, including CNKI, Wanfang, and the China Academic Journals Full-text Database, as well as PubMed and Web of Science. The search terms included " kidney transplant" "flexible ureteroscope" "ureteroscope" and " urinary tract stones" . The search covered all publications available up to December 2023.Result:The 9 recipients had a mean age of 37. 56 years (range: 27–54 years) . The onset time of the kidney stones ranged from 1 to 13 months after kidney transplantation. All recipients were diagnosed with transplanted kidney stones by CT and showed no symptoms of kidney colic at the time of onset. All transplanted kidneys were located in the right iliac fossa, with varying degrees of hydronephrosis. The smallest stone diameter was 0. 5 cm, and the largest was 1. 3 cm. The number of stones ranged from 1 to 3 per recipient; 6 cases had solitary stones, and 3 had multiple stones in the renal pelvis and calyces. In all 9 surgeries, the ureteral orifice of the transplanted kidney was located at the apex of the bladder using ureteroscopy, and a hydrophilic guidewire was inserted into the renal pelvis. A second ureteroscopy was then performed, guiding the ureteroscope tip into the transplanted ureteral orifice in coaxial alignment with the guidewire. A flexible ureteroscope sheath was advanced along the guidewire, followed by the insertion of a digital flexible ureteroscope to perform holmium laser dusting lithotripsy. Stone composition analysis revealed mixed calcium oxalate monohydrate and calcium oxalate dihydrate. The surgical duration ranged from 30 to 75 minutes. No complications such as massive bleeding, septic shock, or rejection occurred. Postoperative review at 1 month, after ureteral stent removal, showed no residual stones in any patient. During 6 months of follow-up, no recurrence of stones was observed, and the renal function of the transplanted kidneys remained stable compared to preoperative levels. A total of 19 related articles were identified through literature review, including 11 in English and 8 in Chinese. Chinese reports on transplanted kidney stones were all from single-center studies with small case numbers. In contrast, foreign meta-analyses and multicenter studies showed that treatments for transplanted kidney stones abroad mainly included antegrade/retrograde ureteroscopy and percutaneous nephroscopy.Conclusion:Ureteroscopy-assisted flexible ureteroscopic lithotripsy is a feasible treatment for transplanted kidney stones. It can effectively protect graft kidney function and has promising clinical application prospects.

Objective:To analyze the drug resistance characteristics and influencing factors in human immunodeficiency virus (HIV)-1 treated patients in Henan Province.Methods:HIV-1 treated patients who had received anti-retroviral therapy (ART) for more than six months and had a viral load >200 copies/mL in the Zhengzhou Sixth People′s Hospital from January to December 2024 were enrolled. Plasma samples were collected. Partial pol region gene sequences and integrase gene sequences of HIV-1 were amplified by reverse transcription nested polymerase chain reaction. The REGA HIV-1 subtype analysis tool was used to determine the subtypes of HIV-1 isolates, and the HIV drug resistance database of Stanford University in the United States was used to analyze the genetic drug resistance mutations and antiviral drug susceptibility. The multivariate logistic regression analysis was used to analyze the factors related to drug resistance. Results:Among 933 HIV-1 treated patients with ART failure, 825 samples were successfully amplified, with the amplification success rate of 88.42%. The overall drug resistance rate was 70.06%(578/825), among which the drug resistance rates of nucleoside reverse transcriptase inhibitor (NRTI), non-nucleoside reverse transcriptase inhibitor (NNRTI), protease inhibitor (PI), and integrase inhibitor (INSTI) were 55.15%(455/825), 64.36%(531/825), 5.70%(47/825), and 2.31%(19/821), respectively. The most common drug resistance mutations included M184I/V (47.88%(395/825)), K103N/S (38.18%(315/825)), and K70E/G/N/Q/R/S/T/del (16.61%(137/825)). Multivariate analysis showed that the baseline CD4 + T cell count <200 cells/μL (adjusted odds ratio ( OR)=2.239, 95% confidence interval ( CI)1.011 to 4.960), an initial 2NRTI+ NNRTI-based treatment regimen (adjusted OR=44.332, 95% CI 5.191 to 378.593), initial 2NRTI+ PI/r (r means ritonavir)-based regimen (adjusted OR=14.391, 95% CI 1.304 to 158.805) and a change in the ART regimen (adjusted OR=5.941, 95% CI 2.373 to 14.878) were independent risk factors for drug resistance (all P<0.05). Conclusions:The drug resistance rate after virological failure in HIV-1 treated patients in Henan Province is relatively high, which is mainly characterized by NNRTI resistance. The baseline immune status and the choice of the initial treatment regimen are important factors affecting the occurrence of drug resistance. The treatment monitoring and drug resistance monitoring should be strengthened.

Approximately 20% to 30% of the global workforce is engaged in shift work. As a significant cause of circadian disruption, shift work is closely associated with an increased risk for periodontitis. Nevertheless, how shift work-related circadian disruption functions in periodontitis remains unknown. Herein, we employed a simulated shift work model constructed by controlling the environmental light-dark cycles and revealed that shift work-related circadian disruption exacerbated the progression of experimental periodontitis. RNA sequencing and in vitro experiments indicated that downregulation of the core circadian protein brain and muscle ARNT-like protein 1 (BMAL1) and activation of the Gasdermin D (GSDMD)-mediated pyroptosis were involved in the pathogenesis of that. Mechanically, BMAL1 regulated GSDMD-mediated pyroptosis by suppressing NOD-like receptor protein 3 (NLRP3) inflammasome signaling through modulating nuclear receptor subfamily 1 group D member 1 (NR1D1), and inhibiting Gsdmd transcription via directly binding to the E-box elements in its promoter. GSDMD-mediated pyroptosis accelerated periodontitis progression, whereas downregulated BMAL1 under circadian disruption further aggravated periodontal destruction by increasing GSDMD activity. And restoring the level of BMAL1 by circadian recovery and SR8278 injection alleviated simulated shift work-exacerbated periodontitis via lessening GSDMD-mediated pyroptosis. These findings provide new evidence and potential interventional targets for circadian disruption-accelerated periodontitis.
Pyroptosis/physiology* ; ARNTL Transcription Factors/metabolism* ; Animals ; Periodontitis/etiology* ; Mice ; Phosphate-Binding Proteins/metabolism* ; Shift Work Schedule/adverse effects* ; Intracellular Signaling Peptides and Proteins/metabolism* ; Mice, Inbred C57BL ; Male ; Disease Models, Animal ; Gasdermins

The paper summarizes Professor YANG Jun's thoughts on clinical treatment with acupuncture and moxibustion. Professor YANG Jun puts forward the "refined mode for diagnosis and treatment of diseases with acupuncture and moxibustion", aiming to improve the capacity of diagnosis and treatment in clinical practice. He advocates that the diagnosis and treatment should be guided by the identification of etiologies, syndromes and meridians; in accordance with regulating the shape/form, balancing yin and yang, and harmonizing the mind; and by means of skillful techniques of acupuncture and moxibustion, simplified selection of acupoints and delicate manipulations. Besides, he stresses on the combination of multiple techniques of acupuncture (such as penetrating technique with long needle, stuck needling by lifting and pulling, and micro-acupuncture systems) with moxibustion techniques (moxibustion for resolving stasis and unblocking collaterals, pressing moxibustion, borneol moxibustion, moxibustion with medicinal plaster) in clinical practice, so as to enhance the therapeutic effects.
Moxibustion/methods* ; Acupuncture Therapy/methods* ; Humans ; China ; Acupuncture Points

Metastatic dissemination is the major cause of death from breast-cancer (BC). Fusobacterium nucleatum (F.n) is widely enriched in BC and has recently been identified as one of the high-risk factors for promoting BC metastasis. Here, with an experimental model, we demonstrated that intratumoral F.n induced BC aggressiveness by transcriptionally activating Epithelial-mesenchymal transition-associated genes. Therefore, the F.n may be a potential target to prevent metastasis. Given the fact that cancer-associated fibroblasts (CAFs) are abundant in BC and located near blood vessels, we report an optogenetic system that drives CAF to in situ produce human antibacterial peptide LL37, with the characteristics of biosafety and freely intercellular trafficking, for depleting intratumoral F.n, leading to a 72.1% reduction in lung metastatic nodules number without affecting the balance of the systemic flora. Notably, mild photothermal treatment was found that could normalize CAF, contributing to synergistically inhibiting BC metastasis. In addition, the system can also simultaneously encode a gene of TNF-related apoptosis-inducing ligand to suppress the primary tumor. Together, our study highlights the potential of local elimination of tumor pathogenic bacteria to prevent BC metastasis.