OBJECTIVE To establish a quantitative analysis of multi-components by single-marker （QAMS） method for simultaneous determination of six flavonoids and two phenolic acids in Perilla frutescens leaves using scutellarin and rosmarinic acid as internal reference substances， and apply this method to determine the contents of eight components in 20 batches of P. frutescens leaves samples from different regions. METHODS Scutellarin served as the internal reference to calculate relative correction factors （RCFs） for scutellarin-7-O-diglucuronide， luteolin-7-O-diglucuronide， apigenin-7-O-diglucuronide， luteolin-7-O- β-D-glucuronide and apigenin-7-O-glucuronide. Rosmarinic acid was employed as the internal reference to determine the RCF for caffeic acid. The contents of the above flavonoids and phenolic acids were calculated with QAMS， and compared with the results of external standard method. RESULTS The eight analytes demonstrated excellent linearity within their respective concentration ranges （r≥0.999 0）. The mean recovery rates for spiked samples ranged from 95.60% to 102.15%， with relative standard deviations （RSDs） of 0.72% to 2.70% （n＝6）. The method exhibited good precision， repeatability， and stability （RSD＜2.50%， n＝6）. Variations in instruments， columns， column temperature， flow rate， and formic acid volume fraction had minimal impact on the RCFs （RSD＜3%， n＝3）. Comparison with the external standard method showed no significant differences in the content of each component across batches， except for caffeic acid in the ZS12 batch （absolute value of RE＜5%， n＝2）. The contents of six CARS-21） flavonoid components in P. frutescens leaves samples varied significantly across different geographic origins， while the content of total flavonoids showed no significant difference. In contrast， the contents of two phenolic acid components and total phenolic acid exhibited significant variation among samples from different regions. CONCLUSIONS The developed QAMS method can simultaneously determine the contents of six flavonoids and two phenolic acids in P. frutescens leaves. It is convenient for detection， highly accurate， and cost-effective. This method is suitable for the quality control of P. frutescens leaves， and the variation of flavonoid and phenolic acid content in samples from different regions provides a reference for the selection of optimal cultivation areas.

BACKGROUND:Upregulation of hedgehog protein signaling can increase the expression of osteoarthritis markers,Runx2,a disintegrin and metalloproteinase with thrombospondin motifs,collagen type X alpha 1,and matrix metalloproteinase 13,while inhibition of hedgehog proteins attenuates the severity of osteoarthritis.It is speculated that osteoarthritic chondrocytes can influence bone formation by affecting osteoblasts through the Indian hedgehog protein(IHH)signaling pathway. OBJECTIVE:To investigate the effect of human osteoarthritic chondrocytes on subchondral osteoblasts. METHODS:Tibial plateau specimens from patients with osteoarthritis were collected.Chondrocytes were extracted using enzymatic digestion,and osteoblasts were extracted using enzymatic pre-digestion + bone block method.Chondrocytes were identified by toluidine blue staining and immunofluorescence and osteoblasts were identified by alkaline phosphatase staining and immunofluorescence.Chondrocytes were cultured in sodium alginate beads to maintain chondrocyte phenotype and co-cultured with osteoblasts.The co-culture system was added with IHH signaling pathway inhibitor(cyclopamine,10 nmol/L)and activator(purmorphamine,10 nmol/L)separately.After 48 hours of co-culture,osteoblasts from each group were collected,mRNA expressions of Gli1,osteoprotegerin,Runx2,parathyroid hormone-related peptide,alkaline phosphatase,receptor activator of nuclear factor-kB ligand(RANKL)and osteocalcin were detected by qRT-PCR,and protein expressions of GLi1,oseoprotegerin and RANKL in osteoblasts were detected by western blot. RESULTS AND CONCLUSION:The mRNA expression levels of GLi1,osteoprotegerin and RUNX2 in osteoblasts were significantly increased,while the mRNA expression levels of parathyroid hormone-related peptide were decreased(P＜0.05)when co-cultured with human osteoarthritic chondrocytes.The mRNA and protein levels of Gli1 were significantly decreased after the addition of IHH signaling pathway inhibitor(cyclopamine)(P＜0.05),and the mRNA and protein levels of Gli1 were significantly increased after the addition of IHH signaling pathway activator(purmorphamine)(P＜0.05).Osteoprotegerin showed the same trend as Gli1 in the experiment.The osteoprotegerin/RANKL ratio followed the same trend as osteoprotegerin.To conclude,human osteoarthritic chondrocytes can promote the expression of Gli1,osteoprotegerin,Runx2 and other proteins in osteoblasts.The upregulation of osteoprotegerin is related to the IHH signaling pathway.Osteoarthritic chondrocytes can up-regulate the expression of osteoprotegerin in osteoblasts through the IHH signaling pathway and thus up-regulate the osteoprotegerin/RANKL ratio,which will contribute to bone formation in subchondral bone.

Objective We aimed to observe the differences in the effects of different acupuncture technique on the endometrium of mice with simple endometrial hyperplasia model and to explore the potential mechanisms. Methods According to the random number tables,32 female C57BL/6J mice were divided into a blank control group,a model group,a quick needle group and a retaining needle group,with 8 mice in each group. A mouse model of simple endometrial hyperplasia was established using bilateral ovariectomy combined with estrogen loading. In the quick needle group,mice were punctured at the bilateral for "Yinbai"(SP1) points and withdrawn immediately,with the treatmeat performed once every other day for a total of 12 times. In the retaining needle group,mice were punctured at the bilateral "Yinbai"(SP1) points and the needles were retained for 15 min each time,with the treatment also performed once every other day for a total of 12 times. After the intervention,samples were collected. HE staining was used to observe morphological changes in the mouse uterine tissue;ELISA was used to detect serum estradiol level;flow cytometry was used to detect the ratio of M1 and M2 macrophages(M1/M2) and immunohistochemical method was used to measure the expression of CD86 and CD206 in uterine tissue;and Western blotting was used to detect the expression of interleukin-13 (IL-13) and interferon-γ(IFN-γ) in uterine tissue. Results The endometrium of mice in the model group showed simple hyperplasia. Compared with the blank control group,the endometrium of the model group was thickened (P<0.01);the level of estradiol in the serum was increased (P<0.01);M1/M2 in uterine tissues was decreased (P<0.01),the expression of CD86 was decreased (P<0.01),and the positive expression of CD206 was increased (P<0.01);and the level of IFN-γ protein expression in uterine tissues was decreased (P<0.01),and the expression of IL-13 protein was increased (P<0.01). Compared with the model group,the endometrial thicknesses of the quick needle group and the retaining needle group were reduced (P<0.05),the levels of estradiol in serum were reduced (P<0.05),M1/M2 in uterine tissues increased (P<0.01),and the reduction of CD206 positive expression,and IL-13 protein expression reduced (P<0.01);the level of CD86 positive expression,IFN-γ protein expression increased (P<0.01). Compared with the quick needle group,IL-13 protein expression increased in the retaining needle group (P<0.01).Conclusion Both quick needle and retaining needle may be through the regulation of the expression of IFN-γ and IL-13,thus prompting the polarization of macrophages from M2 to M1 type,inhibiting the pro-cell proliferative ability and tissue repair ability of M2 type macrophages,thus reducing the degree of endometrial hyperplasia,and the quick needle group was superior to the retaining needle group in regulating the expression of IL-13.

Objective To develop and evaluate the equivalence of the Mandarin test material for tracking of noise tolerance(TNT)test.Methods Six different speech materials were developed(themes including daily life,entertainment,family,festivals,outdoors,and school).Four-minute TNT tests were measured in 21 normal hear-ing subjects using six different test materials.For each session,the tolerable noise level(TNL)and TNT scores were acquired and calculated for 3 time windows(31～240 s,31～120 s,151～240 s).Results Statistic analysis showed significant differences in the TNL(F=43.611,P＜0.05)among the normal hearing listeners.There were statistically significant differences in standardize z-scored TNT scores of the six different materials in the three time windows(P＜0.05).Post-hoc comparisons revealed that all significant differences involved the family and daily life themes.Conclusion Entertainment,festival,outdoors and school themed test materials can serve as the materials of Mandarin tracking of noise tolerance test and can be appied in research and clinical testing.

Objective:To analyze the occurrence characteristics of heat-related diseases in the 19th Hangzhou Asian Games beach volleyball events, strengthen the ability of prevention and early identification of heat-related diseases, and provide reference for the holding of large-scale outdoor events in summer and reasonable allocation of medical resources.Methods:The medical insurance of heat-related diseases of relevant personnel in the beach volleyball competition from September 19 to September 28, 2023 was retrospectively analyzed, and the incidence of heat-related diseases in the personnel involved in Asia was analyzed.Results:During the beach volleyball competition in Ningbo Region of the Hangzhou Asian Games, a total of 103 people were provided with health services in the medical service field (61 people had mild discomfort due to excessive outdoor temperature; Other cold, minor injury, bandage 42 people); Medical services provided 44 times (4 referrals). Among them, 11 cases were sports injury and trauma (29.5%), 11 cases were heat stroke and other related symptoms (25%), 6 cases were sunburn (13.6%), 10 cases were oral diseases of five senses (22.8%), 4 cases were upper respiratory tract infection (9.1%).Conclusions:The holding of large-scale outdoor events in summer should focus on heat-related diseases, and it is necessary to effectively do the corresponding planning work in advance in terms of reasonable allocation of medical resources and targeted training of professionals.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.

Objective:To delineate the histomorphological disparities of subchondral bone between hemophilic arthritis (HA) and osteoarthritis (OA) and to explore the mechanisms underpinning aberrant bone remodeling in HA.Methods:Fifteen male HA patients, aged 32.60±7.58 years (range 22-45), who underwent total knee arthroplasty at the Second Affiliated Hospital of Anhui Medical University from January 2021 to June 2023, were included. All patients had hemophilia A and tested negative for coagulation factor VIII antibodies. Simultaneously, fifteen male OA patients, aged 75.67±5.09 years (range 71-87), also underwent arthroplasty. Tibial plateau bones were extracted for micro-CT, which assessed morphological parameters. Histological changes in the subchondral bone plate (SBP) and trabecular bone were evaluated with HE and Safranin O-Fast Green staining. TRAP staining determined osteoclast differentiation levels, and VEGF-A and Osterix immunohistochemistry gauged angiogenesis and osteoblast differentiation.Results:Micro-CT revealed that HA patients had a BV/TV of 25.14%±0.70% (medial) and 22.31%±0.53% (lateral), Conn.D. of 4.20±0.10 1/mm 3 (medial) and 3.27±0.08 1/mm 3 (lateral), BMD of 0.288±0.006 g/cm 3 (medial) and 0.285±0.004 g/cm 3 (lateral), Tb.Th of 0.257±0.008 mm (medial) and 0.206±0.008 mm (lateral), Tb.N of 0.984±0.043 1/mm (medial) and 0.908±0.026 1/mm (lateral), and Tb.Sp of 0.683±0.008 mm (medial) and 0.808±0.010 mm (lateral). These parameters were significantly lower than those in the OA group except for Tb.Sp, which was higher ( P<0.001). Histological staining indicated that the HA group's SBP thickness was 177.43±6.42 μm (medial) and 117.96±5.08 μm (lateral) with significant differences observed ( P<0.001). TRAP staining showed that TRAP + osteoclasts accounted for 33.4%±3.1% (medial) and 25.1%±2.3% (lateral) in HA subchondral bone, again significantly different ( P<0.001). Immunohistochemical staining revealed VEGFA + cells at 34.1%±5.9% (medial) and 25.9%±3.7% (lateral), and Osterix + cells at 14.6%±1.4% (medial) and 5.8%±1.1% (lateral) in HA patients, differing significantly from the OA group ( P<0.001). Conclusion:The HA group exhibited more extensive subchondral bone destruction, thinner trabeculae, a nearly absent tidemark, higher osteoclast differentiation, increased angiogenesis, and reduced osteoblast differentiation, indicating severe osteoporosis, despite thicker SBP. These findings suggest that targeting abnormal bone remodeling and angiogenesis in HA could retard its progression and provide therapeutic benefits.

Objective:To investigate the expressions of Yes-associated protein 1 (YAP1), YTH domain-containing family protein 2 (YTHDF2) and microRNA-888-3p (miR-888-3p) in patients undergoing radical resection for lung cancer, and their predictive value for short-term prognosis of patients.Methods:A total of 420 lung cancer patients who underwent radical resection for lung cancer in Shanxi Province Cancer Hospital from May 2015 to May 2022 were selected and divided into a modeling set (280 cases) and a validation set (140 cases) in a ratio of 2∶1 by using the computer-generated random number method. The paracancerous normal lung tissues removed by surgery (>5 cm from the edge of the cancer tissues) were used as the control. The relative expressions of YAP1 and YTHDF2 proteins in cancer tissues and paracancerous tissues of the modeling set was detected by Western blotting, and the relative expression of miR-888-3p was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The patients were followed up for 1 year to analyze the prognosis, and the patients in the modeling set who had recurrence, metastasis or cancer-caused death were considered as the poor prognosis group, and the rest of the patients were considered as the good prognosis group. The relative expressions of YAP1 protein, YTHDF2 protein, miR-888-3p and general information of the patients in the two groups were compared; the Cox proportional hazards model was used to analyze the influencing factors of the short-term prognosis of radical resection for lung cancer in the modeling set; a Nomogram prediction model was constructed for the short-term prognosis of radical resection for lung cancer, and its efficacy was verified.Results:The relative expression of miR-888-3p in the modeling set of cancer tissues and paracancerous tissues was 0.49±0.08 and 0.16±0.05, and the difference was statistically significant ( t = 58.53, P < 0.001); the relative expression of YAP1 protein was 0.48±0.06 and 0.12±0.03, and the difference was statistically significant ( t = 89.80, P < 0.001); the relative expression of YTHDF2 protein was 0.23±0.04 and 0.59±0.07, and the difference was statistically significant ( t = 74.72, P < 0.001). After 1 year of follow-up in the modeling set, 32 cases were lost, and 81 (32.66%) of the remaining 248 cases had a poor prognosis (poor prognosis group), while 167 cases (67.34%) were in the good prognosis group. Compared with the good prognosis group, the relative expressions of YAP1 protein and miR-888-3p in cancer tissues was elevated in the poor prognosis group (both P < 0.05), and the relative expression of YTHDF2 protein was decreased ( P < 0.05); the proportions of patients with smoking, coronary artery disease, TNM stage Ⅲ-Ⅳ, maximum tumor diameter ≥ 5 cm, and undifferentiated/lowly differentiated tissues were high in the poor prognosis group, the physical status (PS) score was elevated, and the proportion of patients with regular postoperative radiotherapy was reduced (all P < 0.05). The results of multivariate Cox regression analysis showed that the proportion of patients with smoking ( HR = 2.098, 95% CI 1.143-3.852, P = 0.009), TNM stage ( HR = 2.421, 95% CI 1.350-4.341, P = 0.002), maximum tumor diameter ( HR = 1.883, 95% CI 1.036-3.424, P = 0.011), degree of tissue differentiation ( HR = 3.133, 95% CI 1.590-6.173, P < 0.001), regular postoperative radiotherapy ( HR = 0.417, 95% CI 0.219-0.797, P = 0.003), YAP1 ( HR = 7.043, 95% CI 2.842-17.452, P < 0.001), YTHDF2 ( HR = 0.560, 95% CI 0.418-0.752, P < 0.001), and miR-888-3p ( HR = 4.100, 95% CI 1.789-9.394, P < 0.001) were the independent influencing factors for the short-term prognosis of radical resection for lung cancer. A Nomogram prediction model was constructed, and internal validation showed that the consistency index of the model was 0.822 (95% CI 0.774-0.870); the result of receiver operator characteristic curve showed that the sensitivity of the Nomogram prediction model for predicting the short-term prognosis of radical resection for lung cancer in the modeling set was 97.5% (95% CI 86.8%-99.9%), and the specificity was 82.4% (95% CI 73.0%-89.6%), and the area under the curve (AUC) was 0.943 (95% CI 0.889-0.976); the sensitivity of predicting the short-term prognosis of radical resection for lung cancer in the validation set was 91.53% (95% CI 81.3%-97.2%), and the specificity was 81.5% (95% CI 71.3%-89.2%), and the AUC was 0.922 (95% CI 0.865-0.961). Conclusions:YAP1, YTHDF2 and miR-888-3p are all influencing factors for short-term prognosis of radical resection for lung cancer, and the Nomogram prediction model constructed based on these factors has good predictive efficacy for the short-term prognosis of radical resection for lung cancer.

Objective To quickly observe the tonic effects of Cervi Colla on enriching blood,strengthening bones and anti-aging in zebrafish model.Methods Glyphosate(Gly)was used to construct the erythrocyte injury model in adult zebrafish,and methotrexate(MTX)was used to construct the hematopoietic function injury model of juvenile zebrafish.Prednisolone(Pred)was used to establish the inhibition model of bone formation in zebrafish larvae.The ocular cell apoptosis model of zebrafish larvae was established by dibutyl phthalate(DBP).Results Cervi Colla could improve the Gly-induced abnormal erythrocyte nucleus in adult zebrafish and promote the expression of hematopoietic factors SCL and GATA1.Cervi Colla improved Pred-induced inhibition of bone formation in juvenile zebrafish,and promoted the expression of osteoblast-related gene ALP and Runx2a.The number of ocular cell apoptosis induced by DBP was decreased,and the expression of anti-apoptotic factor Bcl-2 was promoted.Conclusion Cervi Colla has significant effects on protecting erythrocyte,protecting hematopoiesis,protecting bone formation and anti-apoptosis.These effects may be related to replenish blood,anti-osteoporosis,and anti-aging.This study provides a scientific basis for the clinical application of Cervi Colla,and lays a foundation for further development and application.

Intra-abdominal infections (IAI) is common surgical emergencies and have been reported as major contributors to non-trauma deaths in hospitals worldwide. The principles of IAI management included early diagnosis, adequate source control, appropriate antimicrobial therapy, and prompt physiologic stabilization using critical care resources, combined with an optimal surgical approach. In order to facilitate clinical management, establish a global standard and provide guidance for clinicians, the World Society of Emergency Surgery (WSES), the Global Alliance for Infection in Surgery (GAIS), the Surgical Infection Society-Europe (SIS-E), the World Surgical Infection Society (WSIS), and the American Association for the Surgery of Trauma (AAST) worked together to complete an international multi-society document, which provided the evidence-based clinical pathways. Herein, we made a comprehensive interpretation for the clinical pathways combined with the latest domestic and international research developments, aiming to provide evidence for domestic doctors on the diagnosis and treatment of IAI, and ultimately benefit patients.