OBJECTIVE To investigate the mechanism by which Naozhenning granules （NZN） improve learning and memory impairment in a rat model of multiple cerebral concussion （MCC）. METHODS The MCC rat model was established using the closed controlled cortical impact method. The experiment was set up with a blank group （normal saline）， a model group （normal saline）， a piracetam group （positive control group， 0.324 g/kg）， and high-， medium-， and low-dose NZN groups （5.4， 2.7， 1.35 g/kg）， with 11 rats in each group. Drugs or normal saline were administered by gavage once daily for 28 consecutive days. General condition and body weight were monitored throughout the experiment. The sucrose preference rate and novel object recognition index were measured； Evans blue （EB） extravasation in the cerebral cortex was detected； pathological changes of cortical neurons were observed； the levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， interleukin-6 （IL-6）， IL-10， and tumor necrosis factor-α （TNF-α） in the cerebral cortex were determined； and the phosphorylation levels of AMP-activated protein kinase （AMPK）， glycogen synthase kinase 3β （GSK3β）， and Tau protein were detected. RESULTS Compared with the blank group， the model group showed poor mental state， sluggish response to external stimuli， reduced food and water intake， decreased limb flexibility， and disheveled fur. Body weight， sucrose preference rate， and novel object recognition index were significantly decreased （ P ＜0.05）； EB extravasation in the cerebral cortex was significantly increased （ P ＜0.05）， with severe neuronal damage. The positive area ratio of Bax protein， IL-6 and TNF-α levels， and Tau protein phosphorylation level were all significantly increased （ P ＜0.05）， whereas the positive area ratio of Bcl-2 protein， IL-10 level， and AMPK and GSK3β protein phosphorylation levels were significantly decreased （ P ＜0.05）. Compared with the model group， all NZN dose groups showed improvements in general condition and pathological damage， with quantitative indices partially restored， and the differences in quantitative indices in high-dose NZN group were statistically significant （ P ＜0.05）. CONCLUSIONS NZN can effectively improve learning and memory impairment in MCC model rats. The mechanism may be related to activating the AMPK/GSK3β pathway， inhibiting inflammatory response， reducing Tau protein phosphorylation level， and then repairing the neuronal injury.

BACKGROUND: Endoscopic sphincterotomy (EST) is widely used to treat common bile duct stones (CBDS); however, long-term studies have revealed the increasing incidence of recurrent CBDS after EST. Loss of sphincter of Oddi function after EST was the main cause of recurrent CBDS. Reparation of the sphincter of Oddi is therefore crucial. This study aims to investigate the effectiveness and safety of endoclip papilloplasty (ECPP) for repairing the sphincter of Oddi and elucidate its mechanism. METHODS: Eight healthy Bama minipigs were randomly divided into the EST group and the ECPP group at a 1:1 ratio, and bile samples were collected before endoscopy and 6 months later. All minipigs underwent transabdominal biliary ultrasonography for the diagnosis of cholelithiasis 6 months after endoscopy. The biliary microbiota composition and alpha and beta diversity were analyzed by 16S ribosomal RNA gene sequencing. Differential metabolites were analyzed by bile acid metabolomics to explore the predictive indicators of cholelithiasis. RESULTS: Three minipigs were diagnosed with cholelithiasis in the EST group, while none in the ECPP group showed cholelithiasis. The biliary Firmicutes/Bacteroidota (F/B) ratio was increased after EST and decreased after ECPP. The Chao1 and observed species index significantly decreased 6 months after EST ( P  = 0.017 and 0.018, respectively); however, the biliary α-diversity was similar before and 6 months after ECPP. The β-diversity significantly differed in the EST group before and 6 months after EST, as well as in the ECPP group before and 6 months after ECPP (analysis of similarities [ANOSIM]: R  = 0.917, P  = 0.040; R  = 0.740, P  = 0.035; respectively). Glycolithocholic acid (GLCA) and taurolithocholic acid (TLCA) accumulated in bile 6 months after EST. CONCLUSIONS ECPP has less impact on the biliary microenvironment than EST and prevents duodenobiliary reflux by repairing the sphincter of Oddi. The bile levels of GLCA and TLCA may be used to predict the risk of cholelithiasis.
Animals ; Swine, Miniature ; Swine ; Cholelithiasis/prevention & control* ; Sphincterotomy, Endoscopic/methods* ; Sphincter of Oddi/surgery* ; Female ; Male

OBJECTIVE: To establish a three-dimensional finite element model of a digital wire loop space maintainer for the mandible and primary tooth loss, in order to investigate the stress, deformation, and shear force experienced by patients with the loss of the second primary molar when wearing the wire loop space maintainer. METHODS: Cone beam computed tomography (CBCT) scans were performed on the patients to create a digital model of the mandible with the absence of the second primary molar using Mimics 21.0 software. A digital model integrating the crown's retention and the wire loop structure of the full crown and ring wire loop space maintainer was constructed using pediatric space maintainer design software, utilizing three different materials: cobalt-chromium alloy, polyether ether ketone (PEEK), and titanium alloy. In ANSYS Work Beach 2023 R2 software, vertical loads of 70 N, tilted 45° along the long axis of the tooth loads of 70 N, and a 10 N load on the surface of the wire loop were applied to the occlusal surfaces of models 46 and 84, simulating centric and lateral occlusions during chewing with the wire loop space maintainer in place. The stress states of the wire loop space maintainer and supporting teeth were analyzed. RESULTS: Under various loading conditions, the maximum principal stress of the ring wire loop space maintainer was significantly lower than that of the full crown. Stress contour maps indicated that the peak of the maximum principal stress occurred at the junction of the wire loop and crown structure, indicating that this area was more susceptible to fracture. The ring wire loop space maintainer made from PEEK material exhibited the lowest maximum shear stress on the internal organizational surfaces, with equivalent stresses of 23.18 MPa and 36.35 MPa for models 46 and 84, respectively. Stress contour maps demonstrated that the maximum stress on tooth 46 was located at its mesial, while the maximum stress on tooth 84 was situated near the root area on its distal, in contact with the wire loop space maintainer. CONCLUSION In cases of second primary molar loss, wearing the digital ring wire loop space maintainer can effectively distribute stress, and the ring wire loop space maintainer made from PEEK material reduces the stress experienced by supporting teeth to some extent, demonstrating its superiority in clinical application.
Finite Element Analysis ; Humans ; Tooth, Deciduous ; Cone-Beam Computed Tomography ; Space Maintenance, Orthodontic/methods* ; Imaging, Three-Dimensional ; Orthodontic Wires ; Dental Stress Analysis ; Mandible ; Stress, Mechanical

Objective: To explore the effect of HIF-1α on osteogenic-angiogenic coupling response in bone mesenchymal stem cells (BMSCs) and provide new concepts for engineered bone tissue in vitro. Methods: With the approval of the hospital’s experimental animal ethics committee, BMSCs were harvested from Wistar rats. The lentivirus carrying hypoxia-inducible factor-1α (HIF-1α) and empty lentivirus were stably transfected into the third generations of BMSCs to form LV-HIF-1α-BMSCs and LV-BMSCs. Meanwhile, BMSCs without transfection of lentivirus were used as a blank control. Then, the effect of HIF-1α transfection was verified by qPCR and Western Blot. LV-HIF-1α-BMSCs were induced to differentiate into endothelium-like cells (iECs). The morphology was observed by optical microscopy, the differentiation rate was detected by cellular flow CD31, and the Transwell test was used to detect the migration ability. At the same time, LV-HIF-1α-BMSCs and LV-BMSCs were continuously cultured to form osteogenic cell sheets (OCTs), which were stained by alkaline phosphatase on day 14 and alizarin red staining on day 21, and counted for mineralization capacity. Finally, iECs were implanted into OCTs to form prevascularized osteogenic cell sheets (P-OCTs), immunofluorescence CD31 was performed to detect the formation of vascular networks, and the results were recorded on days 1, 3, 7, and 14. Meanwhile, osteopontin (OPN) and osteocalcin (OCN) were detected by western blot to verify their ability for osteogenic differentiation on days 1, 7, and 14. Results: The optimal multiplicity of infection (MOI) for lentiviral transfection was 30, and the transfection efficiency was >80%. The results of qPCR and western blot showed that compared with the LV-BMSCs group and BMSCs group, the LV-HIF-1α-BMSCs group had stable and high expressions of HIF-1α (P<0.05). LV-HIF-1α-BMSCs showed an enhanced ability to differentiate into endothelial cells, with a differentiation rate as high as 91.81%. Transwell assay verified that HIF-1α could recruit iECs in vitro. Alkaline phosphatase staining and alizarin red staining confirmed that OCTs formed by LV-HIF-1α-BMSCs had a statistically significant osteogenic differentiation ability compared with LV -BMSCs control group (P<0.05). When iECs were implanted into the LV-HIF-1α-BMSCs group OCTs to form P-OCTs, iECs substantially proliferated and rapidly fused, and formation of the progressive lumen was revealed by immunofluorescent CD31 staining. The expressions of OPN and OCN were significantly enhanced compared with those of the LV-BMSCs control group; OCN was the highest on day 7, and OPN was the highest on day 1 (P<0.05). Conclusion BMSCs transfected by HIF-1α have good osteogenic-angiogenic effect after induction and differentiation, which provides experimental foundation for optimizing the construction of three-dimensional prevascularized bone tissue.

OBJECTIVE To study the effects of peripheral blood-derived exosomes （Exo） intervened by Naozhenning （NZN） on injury of neuron cells HT22 induced by microglia BV-2 cells. METHODS Wistar rats were selected to prepare peripheral blood- derived Exo intervened by NZN （66.83 g/kg）， referred to as NZN-Exo； peripheral blood-derived Exo intervened by normal saline and piracetam （PLXT， 1.62 g/kg） were prepared using the same method， denoted as KB-Exo and PLXT-Exo respectively， and all Exo were subsequently identified. Meanwhile， BV-2 cells were stimulated with 1 μg/mL lipopolysaccharide （LPS） to prepare LPS- stimulated supernatant， and non-LPS-stimulated supernatant was prepared following the same protocol. HT22 cells were divided into four groups: KB-Exo group （treated with non-LPS-stimulated supernatant+KB-Exo）， model group （treated with LPS-stimulated supernatant+KB-Exo）， PLXT-Exo group （treated with LPS-stimulated supernatant+PLXT-Exo）， and NZN-Exo group （treated with LPS-stimulated supernatant+NZN-Exo）， with the concentration of the corresponding Exo in all groups being 50 μg/mL. After 24 hours of culture， the proliferation of HT22 cells was detected by the CCK-8 assay and EdU assay； the apoptosis of HT22 cells was detected； the microstructure of HT22 cells was observed； the contents of interleukin-1β （IL-1β）， IL-10， nuclear factor-κB （NF- κB）， and tumor necrosis factor-α （TNF-α） in HT22 cells were measured， as well as the expression levels of TNF-α， NOD-like receptor thermal protein domain associated protein 3 （NLRP3）， Caspase-1， B-cell lymphoma-2（ Bcl-2）， and Bcl-2-associated X protein （Bax）. RESULTS KB-Exo， PLXT-Exo and NZN-Exo were successfully prepared， and all Exo exhibited typical cup-shaped contours and membrane-enclosed characteristics. Compared with KB-Exo group， model group showed significantly decreased cell proliferation rates （detected by CCK-8 and EdU）， intracellular IL-10 levels， and Bcl-2 protein expression levels （P＜0.05）； while the cell apoptosis rate， intracellular levels of IL-1β， TNF-α， and NF-κB， as well as the expression levels of NLRP3， TNF-α， Caspase-1， and Bax proteins were significantly increased （P＜0.05）. Additionally， in the model group， the cells showed volume swelling， incomplete cell membrane， nucleolar rupture， significant swelling and deformation of mitochondria， and severe vacuolization. Compared with model group， the above quantitative indicators in the PLXT-Exo group and NZN-Exo group were significantly reversed （P＜0.05）， with large and round cell nuclei， intact nuclear membranes， and reduced mitochondrial vacuolization. CONCLUSIONS Peripheral blood-derived Exo intervened by naozhenning can alleviate the injury of neuronal cells HT22 by inhibiting inflammatory responses and cell apoptosis.

Objective: To determine the clinical efficacy of platelet-rich plasma (PRP) injection combined with warm acupuncture and moxibustion for treating patients with knee osteoarthritis and cold dampness obstruction syndrome. Methods: One hundred and twenty-eight patients with knee osteoarthritis and cold dampness obstruction syndrome who visited the Rehabilitation Department and Orthopedics Department of the Second Affiliated Hospital of Anhui University of Chinese Medicine from January 2023 to March 2024 and who met the inclusion and exclusion criteria were randomly divided into an experimental (n=64) and control group (n=64) using the random number table method. The experimental group was treated with PRP injection combined with warm acupuncture and moxibustion, whereas the control group was treated with normal saline injection combined with warm acupuncture and moxibustion treatment. PRP and normal saline injections were administered once every two weeks, a total of four times. Patients were treated with warm acupuncture and moxibustion once a day, six times a week, for four consecutive weeks. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Traditional Chinese Medicine (TCM) syndrome, visual analog scale (VAS), and Lysholm scores were determined before treatment, at week 4 and week 8 of treatment, and week 16 of follow-up. Serum interleukin-6 (IL-6), matrix metalloproteinase-3 (MMP-3), tumor necrosis factor-α (TNF-α), osteoprotegerin (OPG), bone gla protein(BGP), and cartilage oligomeric matrix protein (COMP) levels were compared between the two groups before and after 8 weeks of treatment. The clinical efficacy and safety indicators between the two groups were also compared. Results: There was no statistical difference in baseline data such as gender, age, disease duration, and body mass index between the two groups of patients. Compared with before treatment, both groups showed decreased WOMAC total and subscale, TCM syndrome total score and symptom scores, and VAS scores, and an increase in Lysholm scores at 4, 8, and 16 weeks after treatment. After treatment, serum IL-6, MMP-3, TNF-α, and COMP levels decreased, whereas serum OPG and BGP levels increased (P<0.05). Compared with the control group, patients in the experimental group showed decreased WOMAC total and subscale, TCM syndrome total score and symptom scores, and VAS scores, and an increase in Lysholm score at 4, 8, and 16 weeks after treatment. Compared with the control group, patients in the experimental group showed decreased serum IL-6, MMP-3, TNF-α, and COMP levels and an increase in serum OPG and BPG levels after treatment (P<0.05). The total effective rate of the experimental group was 91.94%, higher than that of the control group (81.97%; P<0.05). Conclusion PRP injection combined with warm acupuncture and moxibustion can improve various TCM symptoms, improve knee joint function and bone metabolism, and reduce inflammation in patients with knee osteoarthritis and cold dampness obstruction syndrome.

[Purpose]To analyze the incidence trend and age,period,cohort effects of brain and other central nervous system tumors(brain tumor)in Qidong City of Jiangsu Province from 1972 to 2021.[Methods]The incidence data of brain tumor from 1972 to 2021 were collected from the Qidong cancer registry database.The crude incidence rate(CR),age-standardized rate by Chi-nese standard population(ASRC),age-standardized rate by world standard population(ASRW)and average annual percentage change(AAPC)were calculated.The age-period-cohort(APC)model was used to analyze the age,period,and cohort effects of brain tumor incidence in Qidong City from 1972 to 2021.[Results]A total of 2 801 cases of brain tumor occurred in Qidong City from 1972 to 2021,including 1 475 male cases and 1 326 female cases.From 1972-1976 to 2017-2021,the CR increased from 1.81/10 5 to 9.28/10 5,the ASRC increased from 2.07/10 5 to 4.96/10 5,and ASRW increased from 2.00/105 to 4.98/105.From 1972 to 2021 the AAPC of CR,ASRC and ASRW were 3.97％(3.53％for male and 4.37％for female),2.02％(1.66％for male and 2.24％for female)and 2.06％(1.63％for male and 2.24％for female)(all P＜0.001).The Wald's Chi-square test of the APC model showed that there were significant differences in the trends of age,period and birth cohort effect of brain tumor risk(all P＜0.05).The age effect showed that the incidence of brain tumor increased with age,starting from the age group of 45～49 years old and reaching a peak of 25.84/10 5 in age group of 75～79 years old(95％CI:21.17/10 5～31.53/10 5).The period ef-fect showed that the risk of recent period was higher than that in the early period using 1992-1996 period as the reference,reaching the highest in 2012-2016(RR=1.64,95％CI:1.38～1.95).The birth cohort effect showed that the highest risk was in 2017-2021 births cohort(RR=11.17,95％CI:4.26～29.26)using 1952-1956 birth cohort as the reference.[Conclusion]The incidence of brain tumor in Qidong City has been rising;and age,period and cohort are the main influencing factors,suggesting that the middle-aged and elderly population should be the focus of brain tumor prevention and control.

Objective:To analyze the incidence trends of malignant tumors in the elderly population aged over 60 years in Qidong City from 1972 to 2021, as well as to predict the incidence rate for the next decade.Methods:Data were obtained from the Qidong Cancer Registry and Reporting System.The crude incidence rate(CR), age-standardized incidence rate using the Chinese standard population(ASRC), and age-standardized incidence rate based on Segi's world standard population(ASRW)were calculated.Joinpoint regression analysis was conducted using Joinpoint 4.9.1.0 software to determine the average annual percentage change(AAPC)in incidence.Additionally, the ARIMA model implemented in SAS 9.2 software was utilized for time series analysis to forecast incidence trends over the forthcoming 10 years.Results:In Qidong City, a total of 87, 401 malignant tumors were reported in the elderly population.The ASRW increased from 736.85 per 100, 000 in the years 1972-1976 to 1 056.33 per 100, 000 in 2017-2021.Specifically, the ASRW for males rose from 968.56 per 100, 000 to 1 332.75 per 100, 000, while the ASRW for females increased from 550.62 per 100, 000 to 825.44 per 100, 000, with AAPC values of 1.16%, 0.94%, and 1.44% over 50 years(all P<0.001).The incidence trend exhibited an upward trajectory with age, peaking in the 75-79 age group.The AAPC values for the incidence rates in the age groups of 60-64, 65-69, 70-74, 75-79, and those aged over 80 were 0.64%, 0.93%, 0.92%, 2.02%, and 2.44%, respectively(all P<0.001).Among the various cancers, lung cancer, which ranked first, saw an increase in ASRW from 100.87 per 100, 000 in 1972-1981 to 248.84 per 100, 000 in 2012-2021.In contrast, gastric cancer, ranked second, decreased from 216.23 per 100, 000 in 1972-1981 to 103.54 per 100, 000 in 2012-2021.Liver cancer, ranked third, fluctuated from 113.47 per 100, 000 in 1972-1981 to 125.13 per 100, 000 in 2012-2021.Colorectal cancer, ranked fourth, increased from 40.06 per 100, 000 in 1972-1981 to 123.47 per 100, 000 in 2012-2021, while esophageal cancer, ranked fifth, decreased from 63.42 per 100, 000 in 1972-1981 to 28.65 per 100, 000 in 2012-2021.The AAPC values over 50 years for these cancers were 2.25%, -1.89%, 0.36%, 3.13%, and -1.86%, respectively(all P<0.05).Projections indicate that by 2031, the incidence of malignant tumors among the elderly population in Qidong will reach 1 253.84 per 100, 000, with estimates of 1 566.67 per 100, 000 for males and 983.14 per 100, 000 for females. Conclusions:The incidence of malignant tumors among the elderly population in Qidong City is increasing.Common types of cancer in this demographic include lung cancer, gastric cancer, liver cancer, colorectal cancer, and esophageal cancer.Notably, lung cancer, liver cancer, and colorectal cancer are on the rise and should be prioritized in cancer prevention and control efforts.

Objective:A retrospective study was conducted to investigate the safety of Percutaneous Endoscopic Gastrostomy(PEG)in elderly patients with Amyotrophic Lateral Sclerosis(ALS)and to identify the factors influencing major complications.The aim is to establish a more robust foundation for optimizing both the procedure and its timing, thereby enhancing the evidence base for refining the surgical approach.Methods:We retrospectively collected clinical data from elderly ALS patients(≥60 years old)who underwent PEG at Peking University Third Hospital between January 2006 and January 2024.This dataset includes information on general health conditions, the progression of ALS, comorbidities, surgical details, and related complications.Additionally, we analyzed the postoperative complications experienced by the included patients, focusing specifically on the risk factors associated with aspiration pneumonia.Results:A total of 140 patients were included in the study, with a mean age of 68.2±5.9 years.Among these, 69 were male(49.3%)with a mean age of 68.4±6.5 years, and 71 were female(50.7%)with a mean age of 67.7±6.0 years.Successful outcomes were observed in 139 cases(99.3%).One case was not completed due to respiratory arrest caused by intraoperative aspiration.Postoperative complications occurred in 36 cases(25.7%), which included 9 cases of wound infection(6.4%), 1 case of abdominal infection(0.7%), 21 cases of aspiration pneumonia(15.0%), 1 case of local fistula leakage(0.7%), 2 cases of transient fever(1.4%), and 2 cases of death during hospitalization(1.4%).Both univariate and multivariate analyses indicated that an onset duration of ≤1 year( P=0.020)and a half-sitting position( P=0.022)significantly influenced the occurrence of aspiration pneumonia, acting as protective factors( β<0, OR<1). Conclusions:PEG is a safe method for providing enteral nutrition to elderly patients with ALS.While most complications associated with the procedure are mild and can be managed, Aspiration pneumonia remains a common and serious complication.However, early surgical intervention and the use of a semi-seated position during the procedure can help reduce the incidence of aspiration pneumonia.

Objective: To investigate the prevalence and associated factors of spinal curvature abnormalities among primary and secondary school students in the Guangxi Zhuang Autonomous Region, so as to provide a scientific basis for the prevention and control of such abnormalities. Methods: From September to November 2023, adopting a stratified cluster random sampling method, spinal curvature screenings and questionnaire surveys were conducted among 168 931 students from grade 4 of primary school to grade 12 of high school in 111 districts and counties across 14 cities in Guangxi. Chi square tests and binary Logistic regression analysis were used to analyze influencing factors of spinal curvature abnormalities. Results: In 2023, the detection rate of poor posture among students above grade 4 in Guangxi was 4.24% , and the detection rate of spinal curvature abnormalities was 2.13%. The detection rate was higher among urban students (2.84%) than rural students (1.66%), boarding students (2.61%) than non-boarding students (1.60%), and high school students (3.16%) than junior high (2.45%) and primary school students (1.15%), and the differences were statistically significant ( χ 2=269.85, 221.44, 565.10, P <0.01). A trend of increasing detection rates with higher grade levels was observed ( χ 2 trend =617.63, P <0.01). Binary Logistic regression analysis indicated that students without boarding at school ( OR =0.82, 95% CI =0.75-0.90), engaging in high-intensity physical activity for over 60 min per day ≥5 days per week ( OR =0.90, 95% CI =0.82-0.98), and adequate sleep ( OR =0.87, 95% CI =0.81-0.94) had lower risks of detecting spinal curvature abnormalities ( P <0.05). Conclusions The prevalence of spinal curvature abnormalities increases with grade level among primary and secondary school students in Guangxi. Regular moderate-to-vigorous physical activity demonstrates protective effects against spinal abnormalities.