ObjectiveA mouse model of vaccinia virus Tiantan strain (VTT)-induced encephalopathy was developed using AG6 mice. MethodsVTT was amplified by infecting Vero cells at a multiplicity of infection (MOI) of 0.01, followed by concentration and titration. After 72 h of incubation, virus-containing cells were collected and subjected to concentration. The concentrated viral suspension was serially diluted (10-fold dilutions) and added to 6-well plates containing confluent Vero cell monolayers for plaque assay. The number of plaques formed in each well was counted, and the virus titer was calculated based on the dilution factor. Fourteen 5-6-week-old AG6 mice (half male and half female, housed separately by sex) were randomly divided into a control group (n=3, PBS), a low-dose group (n=6, 1×10⁵ PFU), and a high-dose group (n=5, 5×10⁵ PFU). The mice were anesthetized by isoflurane inhalation and then infected via intranasal instillation. The mental state of the mice in each group was observed daily, and the body weight and mortality were recorded. On day 13 post-infection, 2% Evans Blue (4 mL/kg body weight) was administered via tail vein injection to assess blood-brain barrier (BBB) disruption. Subsequently, brain tissue samples were collected for immunofluorescence analysis to evaluate the activation of astrocytes and microglia. ResultsThe titer of purified VTT was 1×10⁷ PFU/mL. Compared with the control group, mice in the low-dose group showed no significant change in body weight, and no lethality was observed. In contrast, mice in the high-dose group exhibited significant weight loss starting on day 5 post-infection (P<0.05), accompanied by lethality. On day 13 post-infection, no Evans Blue extravasation was detected in the brain tissues of the low-dose group, while the olfactory bulb region of the high-dose group displayed distinct blue staining, indicating disruption of the BBB. Immunofluorescence analysis revealed no significant proliferation of astrocytes and microglia in the olfactory bulb region of the low-dose group on day 13 post-infection. In contrast, marked activation of glial cells was observable in the high-dose group. ConclusionAn animal model of VTT-induced encephalopathy in AG6 mice is successfully established, characterized by BBB disruption and reactive gliosis specifically localized to the olfactory bulb region, manifested as astrocytic and microglial proliferation.

Objective To observe and analyze the clinical efficacy of Hewei Jiangni Formula in treating patients with non-erosive gastroesophageal reflux disease(NERD)and cold-heat complex syndrome and explore its regulatory mechanism on visceral hypersensitivity.Methods Sixty patients with NERD and cold-heat complex syndrome diagnosed at the Gastroenterology Clinic of Dongfang Hospital,Beijing University of Chinese Medicine from January 2021 to Decemer 2023,were randomly divided into the Hewei Jiangni Formula group and omeprazole group,with 30 patients per group.The Hewei Jiangni Formula group was treated with Hewei Jiangni Formula and omeprazole enteric-coated tablets placebo,whereas the omeprazole group was treated with omeprazole enteric-coated tablets and Hewei Jiangni Formula placebo.The treatment period was scheduled to last for 8 weeks.The primary outcome indicators(Gastroesophageal Reflux Disease Quality of Life Questionnaire(GERD-Q)scale score:response rate,total score,and single symptom score)and secondary outcome indicators(traditional Chinese medicine clinical scores scale)were recorded before and after treatment in both groups.Ten healthy individuals were recruited from the Physical Examination Center of Dongfang Hospital,Beijing University of Chinese Medicine,as the healthy control group.The expression levels of mast cell tryptase(MCT),protease-activated receptor 2(PAR2),transient receptor potential vanilloid 1(TRPV1),substance P(SP),calcitonin gene-related peptide(CGRP),5-hydroxytryptamine(5-HT),and 5-hydroxytryptamine 3 receptor(5-HT3R)were recorded.These indicators were then used for the following comparisons:the NERD patients vs.the healthy group before treatment;the Hewei Jiangni Formula group vs.the omeprazole group after treatment;the intra-group comparisons within both NERD groups.Changes in serum levels of these indicators were observed before and after treatment in both NERD groups.Results According to the GERD-Q scores before and after treatment,the effective rate was 90.00%in the Hewei Jiangni Formula group and 86.67%in the omeprazole group,with no significant differences.Both groups exhibited improved symptoms of heartburn,regurgitation,upper abdominal pain,and sleep disorders caused by heartburn or regurgitation(P<0.05);however,neither group exhibited significantly improved nausea(P>0.05).Before treatment,MCT,PAR2,TRPV1,SP,CGRP,5-HT,and 5-HT3R expression levels in both NERD groups were significantly higher than those in the healthy control group(P<0.05).After treatment with Hewei Jiangni Formula or omeprazole enteric-coated tablets,the expression levels of these indicators decreased in both groups(P<0.05).The omeprazole group was superior to Hewei Jiangni Formula in reducing 5-HT3R expression(P<0.05).Conclusion Hewei Jiangni Formula significantly improves the symptoms of patients with NERD and cold-heat complex syndrome,with efficacy comparable to that of omeprazole enteric-coated tablets.It is superior to omeprazole in improving symptoms of spleen yang deficiency,such as loss of appetite,fatigue,borborygmus with loose stools,and cold extremities.Hewei Jiangni Formula reduces MCT,PAR2,TRPV1,SP,CGRP,5-HT,and 5-HT3R expression levels,regulates related signaling pathways,and alleviates visceral hypersensitivity.

Objective To observe and analyze the clinical efficacy of Hewei Jiangni Formula in treating patients with non-erosive gastroesophageal reflux disease(NERD)and cold-heat complex syndrome and explore its regulatory mechanism on visceral hypersensitivity.Methods Sixty patients with NERD and cold-heat complex syndrome diagnosed at the Gastroenterology Clinic of Dongfang Hospital,Beijing University of Chinese Medicine from January 2021 to Decemer 2023,were randomly divided into the Hewei Jiangni Formula group and omeprazole group,with 30 patients per group.The Hewei Jiangni Formula group was treated with Hewei Jiangni Formula and omeprazole enteric-coated tablets placebo,whereas the omeprazole group was treated with omeprazole enteric-coated tablets and Hewei Jiangni Formula placebo.The treatment period was scheduled to last for 8 weeks.The primary outcome indicators(Gastroesophageal Reflux Disease Quality of Life Questionnaire(GERD-Q)scale score:response rate,total score,and single symptom score)and secondary outcome indicators(traditional Chinese medicine clinical scores scale)were recorded before and after treatment in both groups.Ten healthy individuals were recruited from the Physical Examination Center of Dongfang Hospital,Beijing University of Chinese Medicine,as the healthy control group.The expression levels of mast cell tryptase(MCT),protease-activated receptor 2(PAR2),transient receptor potential vanilloid 1(TRPV1),substance P(SP),calcitonin gene-related peptide(CGRP),5-hydroxytryptamine(5-HT),and 5-hydroxytryptamine 3 receptor(5-HT3R)were recorded.These indicators were then used for the following comparisons:the NERD patients vs.the healthy group before treatment;the Hewei Jiangni Formula group vs.the omeprazole group after treatment;the intra-group comparisons within both NERD groups.Changes in serum levels of these indicators were observed before and after treatment in both NERD groups.Results According to the GERD-Q scores before and after treatment,the effective rate was 90.00%in the Hewei Jiangni Formula group and 86.67%in the omeprazole group,with no significant differences.Both groups exhibited improved symptoms of heartburn,regurgitation,upper abdominal pain,and sleep disorders caused by heartburn or regurgitation(P<0.05);however,neither group exhibited significantly improved nausea(P>0.05).Before treatment,MCT,PAR2,TRPV1,SP,CGRP,5-HT,and 5-HT3R expression levels in both NERD groups were significantly higher than those in the healthy control group(P<0.05).After treatment with Hewei Jiangni Formula or omeprazole enteric-coated tablets,the expression levels of these indicators decreased in both groups(P<0.05).The omeprazole group was superior to Hewei Jiangni Formula in reducing 5-HT3R expression(P<0.05).Conclusion Hewei Jiangni Formula significantly improves the symptoms of patients with NERD and cold-heat complex syndrome,with efficacy comparable to that of omeprazole enteric-coated tablets.It is superior to omeprazole in improving symptoms of spleen yang deficiency,such as loss of appetite,fatigue,borborygmus with loose stools,and cold extremities.Hewei Jiangni Formula reduces MCT,PAR2,TRPV1,SP,CGRP,5-HT,and 5-HT3R expression levels,regulates related signaling pathways,and alleviates visceral hypersensitivity.

Objective:To explore the effect of personalized training based on early start Denver model (ESDM) combined with applied behavior analysis (ABA) in children with autism spectrum disorder (ASD) .Methods:From January 2021 to January 2023, convenience sampling was used to select 90 children with ASD who received treatment at the Shaoxing Seventh People's Hospital and their caregivers as research subjects. The children were divided into two groups using a random number table method in a 1∶1 ratio. The control group received routine rehabilitation, while the observation group was treated with personalized training based on ESDM combined with ABA. Both groups were intervened for six months. Children's Autism Rating Scale (CARS), Autism Behavior Checklist (ABC), Gesell Developmental Schedule (GDS), Sign-Significate Relation (S-S) Language Development Delay Assessment, Social Life Ability Scale for Infant-Junior Middle School Students (S-M), and Modified Barthel Index (MBI) were used to evaluated the effects of intervention on two groups of ASD children before and after intervention.Results:After six months of intervention, the CARS and ABC scores of ASD children in the observation group were lower than those in the control group, and the scores of GDS dimensions and total developmental quotient, S-S Language Development Delay Assessment score, S-M score, and MBI score were higher than those in the control group, and the differences were statistically significant (all P<0.05) . Conclusions:Personalized training based on ESDM combined with ABA has a good effect, which can improve loneliness symptoms of children with ASD, alleviate cognitive, language, and behavioral dysfunction, enhance social adaptation ability and activities of daily living.

Objective To assess effect of ultrasound-guided erector spinae plane block(ESPB)combined with patient controlled intravenous analgesia(PCIA)on postoperative analgesia after cesarean section.Methods A total of 120 full-term singleton pregnant women who underwent cesarean section under spinal anesthesia at Jiaxing Maternal and Child Health Care Hospital from May 2022 to August 2023 were selected.Participants were randomly divided into three groups using a random number table:Group E(ESPB combined with PCIA),group T[transversus abdominis plane(TAP)block combined with PCIA],and control group(PCIA alone),with 40 women in each group.Visual analogue scale(VAS)and Bruggrmann comfort scale(BCS)scores during rest and coughing were recorded at 4h,8h,12h,24h,and 48h postoperatively.Number of effective PCIA presses,total sufentanil dosage,proportion of rescue analgesia and maternal satisfaction were also documented within 48h.Additionally,adverse reactions and neonatal outcomes were observed within the same 48h period.Results Postoperatively,VAS scores for rest and coughing in group E at 8h,12h,24h were significantly lower than those in group T,those in two groups were lower than those in control group(P＜0.05).Postoperatively at 8h,12h,and 24h,BCS scores in group E were significantly higher than those in group T,with both higher than those in control group(P＜0.05).Within 48h after surgery,the number of effective PCIA presses,proportion of rescue analgesia and total sufentanil dosage in group E were lower than those in group T,both lower than those in control(P＜0.05).Moreover,maternal satisfaction score in group E was significantly higher than that in group T,that in two groups was significantly higher than that in control group(P＜0.05).Within 48h after surgery,there were no significant differences in adverse reactions or neonatal outcomes among the three groups(P＞0.05).Conclusion Ultrasound-guided ESPB combined with PCIA outperforms TAP block combined with PCIA,with reducing analgesic dose and enhancing maternal satisfaction and comfort.

Objective: To investigate the willingness to receive measles-containing vaccine (MCV) and its influencing factors among healthcare workers in the Yangtze River Delta region, so as to provide the evidence for improving the measles-containing vaccination rate Methods: Healthcare workers were sampled from 19 medical institutions in each of Shanghai Municipality, Jiangsu Province, Zhejiang Province and Anhui Province for questionnaire surveys using a multi-stage stratified convenience sampling methods from July 2020 to March 2021. Participants' gender, age, educational level, professional title, measles-containing vaccination, awareness of MCV and willingness to receive MCV were collected, and the factors affecting the willingness to receive MCV were identified among healthcare workers using a multivariable logistic regression model. Results: Totally 1 403 questionnaires were allocated, and 1 394 valid questionnaires were recovered, with an effective recovery rate of 99.36%. The respondents included 327 men and 1 067 women, with a male to female ratio of 1∶3.26, and 64.35% (897) were at ages of 31 to 50 years. There were 1 005 respondents with a bachelor degree (72.09%), 765 with middle and senior professional titles (54.88%), 676 with a history of measles-containing vaccination (48.49%), 1 176 with willingness to receive MCV (84.36%) and 218 without willingness to receive MCV due to convenience of vaccination (30.73%) and cost (19.27%). Multivariable logistic regression analysis showed that region (Zhejiang, OR=1.613, 95%CI: 1.054-2.470; Anhui, OR=2.058, 95%CI: 1.259-3.363), and no history of measles (OR=2.219, 95%CI: 1.302-3.781) were factors improving the willingness to receive MCV among healthcare workers, and hospital level (secondary, OR=0.483, 95%CI: 0.306-0.763; tertiary, OR=0.251, 95%CI: 0.160-0.394), history of measles-containing vaccination (no, OR=0.262, 95%CI: 0.172-0.399; unknown, OR=0.386, 95%CI: 0.266-0.559), and unawareness of MCV knowledge (OR=0.208, 95%CI: 0.081-0.536) were factors inhibiting the willingness to receive MCV among healthcare workers. Conclusions The willingness to receive MCV correlates with region, history of measles, hospital level, history of measles-containing vaccination and awareness of MCV knowledge among healthcare workers in the Yangtze River Delta region.

Objective:To investigate the clinical effect of free fibula flap transplantation in repairing the defect of mandibular osteoradionecrosis (ORN).Methods:A total of 151 mandibular ORN patients undergoing free fibular flap transplantation were selected from August 2005 to September 2020 in the Department of Oral and Maxillofacial Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University. Among them, 109 patients were males and 42 patients were females, aged (54.1±10.1) (ranged 31-85) years old. The clinical data of the patients was collected and the survival rate of the flaps and postoperative function were calculated to evaluate the surgical efficacy. The χ 2 test was used for difference analysis. Results:Among the 151 patients, mandibular ORN caused by radiotherapy for nasopharyngeal carcinoma accounted for 79.5% (120/151). The average time for mandibular ORN appeared was 5(6) years after radiotherapy. Facial artery [57.2%(87/152)] and superior thyroid artery [32.9%(50/152)] were the main anastomotic arteries in the recipient area. There was no significant difference in the necrosis rates of the two flaps [10.3%(9/87) and 12.5% (5/50), respectively, P=0.949]. The main anastomotic veins in the recipient area were the external jugular vein [48.4%(135/279)] and the common facial vein [26.5%(74/279)]. Twenty-five cases (16.6%) had one vein anastomosed, and 126 cases (83.44%) had two veins anastomosed. There was no significant difference in the flap necrosis rate between the two conditions [20.0%(5/25) and 7.1%(9/126), respectively, P=0.100]. Ninety-seven cases (64.2%) used the peroneal musculocutaneous-fascia composite flap to repair the maxillofacial soft and hard tissue defects. Thirteen cases (8.6%) underwent the restorations with digital virtual surgery design, of which 5 cases were repaired with dental implants at the same time. After the operations, lower respiratory tract infection occurred in 17 patients (11.3%), and upper respiratory tract obstruction occurred in 3 cases (2.0%). The survival rate of the flap after operation was 90.7% (136/151), and 21 patients (13.9%) had flap vascular crisis. Delayed healing of maxillofacial wounds occurred in 33 cases (21.9%). After 3 to 24 months of follow-ups, 110 patients (76.9%) had no fistula inside/outside the oral cavity, 118 patients (82.5%) had an improvement in opening mouth of increasing (≥0.5 cm) after surgery, 135 patients (94.4%) had pain relief, 97 cases (67.8%) could eat normal diet, semi-liquid or soft food, and 137 cases (95.8%) were satisfied or basically satisfied with the treatment effects. Conclusions:The free fibular flap transplantation is an effective method to repair mandibular ORN defects. Preoperative vascular assessment is helpful for the selection of recipient vessels. Facial artery, superior thyroid artery, external jugular vein and common facial vein can be used as the main recipient vessels. The repair of the peroneal musculocutaneous-fascia composite flap facilitates the closure of internal and external fistulas. Digital technology can help to restore the maxillofacial shape more accurately, improve the patient′s occlusal and chewing function and enhance the quality of life of mandibular ORN patients.

Objective : To analyze the value of virtual surgical planning in the surgical treatment of osteoradionecrosis of the mandible and to provide a reference for clinical practice. .Methods : From September 2017 to June 2018, 13 patients with mandibular osteoradionecrosis were evaluated preoperatively using the 3D virtual surgery software CMF Proplan 2.0. The surgical guide was designed and 3D printed. Bone resection, fibula shaping and bone graft localization were completed during the operation. In some cases, implants were implanted at the same time, and denture restoration was completed 3 to 6 months after surgery. Patients’ general information, perioperative data, and efficacy evaluation were analyzed. Results: All patients underwent surgery successfully. The survival rate of the free fibula musculocutaneous flap was 100% (13/13), and one patient had complications (partial necrosis at the edge of the flap). The follow-up period was 7 to 15 months, and the median time was 10 months. All patients achieved a healing effect. The number of cases with an increase in mouth opening ≥ 1 cm, 0.5 cm ≤ mouth opening increase < 1 cm, and mouth opening increase < 0.5 cm were 5, 6, and 2, respectively. An imaging examination showed that 12 patients had good bone healing, and 1 patient did not completely heal 7 months after operation. The denture restoration was 92.3% (12/13), of which 3 cases were implanted and repaired at the same time. The average chewing efficiency was 56.11% ± 7.12% (42.03%-67.83%). Conclusion Virtual surgical planning is an effective method for the surgical treatment of mandibular osteoradionecrosis, which can reduce the risk of surgery and more effectively perform mandibular shape and function repair.

OBJECTIVE Disturbance of K+ ion balance in inner ear is associated in age-related hearing loss. Our study is to investigate the role of NKCC1 and Na-K-ATPase in cochlea and auditory function regulated by with different expression of NKCC1 and Na-K-ATPase. METHODS Auditory threshold of young or old C57BL/6J mice was measured by auditory brainstem response(ABR). The expression of NKCC1 and Na-K-ATPase in mice cochlea were evaluated by reverse transcription polymerase chain reaction(RT-PCR) and western blotting. Furosemide and Ouabain were applied in vivo to inhibit NKCC1 and Na-K-ATPase in C57BL/6J mice. RESULTS C57BL/6J mice developed hearing loss at 12M by ABR threshold shifting to (75±10), (78±26) and (81±14)dB SPL at frequencies of 8, 16 and 32 kHz; PCR showed that the relative expression of NKCC1 and Na-K-ATPase mRNA in the aged group decreased, which were 0.52±0.06 and 0.35±0.04 times higher than those in the young control group, the difference was statistically significant(t =7.466 and 16.11, all P＜0.05). WB showed that relative expression of NKCC1 and Na-K-ATPase protein level in the aged group decreased by 0.79±0.02 and 0.68±0.05 times as much as that of the young control group, the difference was statistically significant(t =8.857 and 6.771, P all＜0.05). After applied with Furosemide and Ouabain to suppress the two ion transporters, the ABR threshold increased to (50±17), (53±21), (55±17)dB SPL and (56±6), (70±17), (73±6)dB SPL at frequencies of 8, 16 and 32 kHz. CONCLUSION In vivo experiment of C57BL/6J suggested that NKCC1 and Na-K-ATPase might be related to age related hearing loss.

Objective: To establish a TaqMan-MGB probe-based real-time fluorescence RT-PCR assay for avian influenza H5N6 virus used in rapid diagnosis for suspected cases and surveillance for outer environment of live poultry markets. Methods: Based on the conservative sequences of avian influenza H5N6 virus for HA and NA gene published on GenBank, specific primers and TaqMan-MGB probes were designed to develop and optimize for the dual real-time RT-PCR assay. Specificity, sensitivity, repeatability and comparison tests were carried out. Results: This dual real-time RT-PCR detection can be completed within 80 minutes. There was no cross-reaction with other subtypes of influenza virus and common respiratory pathogens. The minimum detection limit could be up to 10 copies/reaction. The correlation coefficient of standard curve for the gene of H5 and N6 were 0.999 and 0.993, and the coefficients of variation for cycle threshold were range from 0.151%-0.549%and 0.213%-0.575%, respectively. The positive and negative coincidence rates of the validation test were 100%. Conclusions This TaqMan-MGB probe-based dual real-time RT-PCR for avian influenza H5N6 virus was rapid, specific and sensitive. It will have a good use in early emergency detection of suspected cases and continuous monitoring of external environment in live poultry trade market.