Zhishi Shaoyaosan is the 34^th prescription in the Catalogue of Ancient Classic Formulas （Second Batch） published by the National Administration of Traditional Chinese Medicine in 2023. It is widely used in clinical practice and has a definite curative effect. However， there is currently a lack of its ancient literature analysis and textual research， and there is no corresponding Chinese patent medicine preparation. By consulting and combing the relevant ancient books of traditional Chinese medicine， this paper analyzes and conducts textual research of the origin， composition， measurement， administration， and efficacy of Zhishi Shaoyaosan. The results show that Zhishi Shaoyaosan is derived from Essentials from the Golden Cabinet written by Zhang Zhongjing in the Eastern Han Dynasty. It is mainly recorded in the name of Zhishi Shaoyaosan in the literature of the past dynasties. The prescription is composed of Aurantii Fructus Immaturus and Paeoniae Radix Alba. The processing method is stir-frying Aurantii Fructus Immaturus to scorch and using raw Paeoniae Radix Alba. The dose of the prescription recorded in the ancient books is mainly an equal amount of Aurantii Fructus Immaturus and Paeoniae Radix Alba in one square-cun spoon， taken three times a day， which is converted into a modern dose of 1.5 g each time （0.75 g Aurantii Fructus Immaturus and 0.75 g Paeoniae Radix Alba each time）. The components of the prescription are ground into powder and taken with barley porridge， three times a day. The efficacy is to break stagnated Qi， harmonize blood， and relieve restlessness and pain. It is mainly used to treat postpartum abdominal pain， acute pelvic inflammatory disease， acute cholecystitis and intestinal diseases， stroke sequelae， and other diseases. This study combs and analyzes the ancient literature recording Zhishi Shaoyaosan and clarifies the key information of the prescription， which provides a basis for promoting the research and development of its patent medicine.

Objective: To explore the relationship of physical activity (PA) and sedentary behavior (SB) with cardiorespiratory fitness (CRF) among middle schoold students in Tibet, so as to provide empirical references for improving the cardiorespiratory fitness and health levels of adolescents in Tibet. Methods: From August to December 2020, 1 225 junior and senior high school students were selected from 2 middle schools in Lhasa, Tibet Autonomous Region, using the stratified cluster random sampling method. Triaxial accelerometers were used to evaluate PA and SB behaviors, and the 20 meter shuttle run was employed to assess CRF among the middle school students. Isochronous substitution modeling was used to analyze the associations of SB, low intensity physical activity (LPA), and moderate vigorous physical activity (MVPA) with CRF, and the saturation threshold effect in the dose response relationship between MVPA and CRF was analyzed through restricted cubic spline and two stage linear regression. Results: After adjusting for covariates such as gender, body mass index and sleep quality score, isotemporal substitution analysis showed that among junior high school students aged 13-15, replacing 30 minutes of SB ( B =1.73) or LPA ( B =2.38) with MVPA were positively associated with CRF (both P <0.05). Among senior high school students aged 16-18, replacing SB ( B =0.99) or LPA ( B =1.38) with MVPA were also positively associated with CRF (both P <0.05). Restricted cubic spline and two piecewise linear regression analyses indicated that only middle school girls aged 13-18 exhibited a saturation threshold effect between MVPA and CRF (logarithmic likelihood ratio test=0.03), with the optimal CRF improvement observed at 60 minutes of MVPA per day ( B=0.13, P ＜ 0.01). Conclusions Reducing SB and LPA while increasing MVPA can improve CRF in Tibetan middle school students. To maximize CRF improvement, middle school girls should engage in at least 60 minutes of MVPA daily.

RNA methylation is a common epigenetic post-transcriptional modification with various patterns,such as N1-methyladenosine(m1A),5-methylcytidine(m5C)and N6-methyladenosine(m6A).RNA methylation can perform specific biological functions in corresponding targets,dynamical and reversible,thus bridging exogenous environmental factors and different disease outcomes.Exogenous chemicals can induce oxidative stress,inflammation,autophagy and cell cycle disorders,which are reg-ulated by specific RNA methylation modifications and bring about epigenetic toxic effects.These altera-tions act as new key molecular events during cellular senescence and the development of aging and age-related disorders.The correlation between RNA methylation and cell senescence will provide a new line of thought for prevention of and interventions in aging.

BACKGROUND:Mesenchymal stem cells possess characteristics such as rapid renewal,targeted homing,tissue repair,and immune regulation,which provide potential for the treatment of inflammatory diseases.In most inflammatory diseases,interleukin-1β is highly expressed.Both exogenous and endogenous mesenchymal stem cells unavoidably exist in an environment with high interleukin-1β concentration. OBJECTIVE:To study the interaction of interleukin-1β with mesenchymal stem cells in inflammatory environment and the mechanism of its influence on the migration and adhesion of mesenchymal stem cells to provide a theoretical basis for adjusting stem cell therapy strategies. METHODS:The first author searched for studies involving interleukin-1β enhancing migration and adhesion of mesenchymal stem cells by computer on CNKI,WanFang,VIP,PubMed,and Web of Science using search terms"interleukin-1β,mesenchymal stem cell,nuclear factor-κB,MAPK,ERK,p38,migration,adhesion"in Chinese and English.The literature tracing method was also used to search for some of the literature.Finally,65 articles were included in the review analysis. RESULTS AND CONCLUSION:(1)In the inflammatory environment,interleukin-1β can regulate the migration and adhesion ability of mesenchymal stem cells.This effect may be achieved by recruiting IRAK1 through interleukin-1RI and then activating TAK1 and IKK in turn.After IKK phosphorylation,nuclear factor-κB and ERK signaling pathways are activated or CXCR expression is upregulated through the p38 pathway to promote mesenchymal stem cell migration and adhesion.However,further standardized research needs to be carried out based on the genetic background of mesenchymal stem cells,the dose and processing time of interleukin-1β.(2)In vitro experiments using pre-stimulated mesenchymal stem cells with interleukin-1β can change the survival environment of mesenchymal stem cells and alter their secretion factors to make them develop towards a more anti-inflammatory direction.On the other hand,under the premise of producing higher levels of anti-inflammatory and pro-nutrient factors,extracted mesenchymal stem cell exosomes can exert anti-inflammatory effects.(3)It has been observed in various animal disease models that pre-stimulating mesenchymal stem cells with interleukin-1β regulates their immune regulation ability,thereby affecting the development and outcome of inflammation.However,this is limited to preclinical basic research only;further verification on efficacy and safety of stem cell therapy with interleukin-1β pre-treated mesenchymal stem cells is required in clinical settings.

Cancer is one of the leading causes of morbidity and mortality worldwide. Due to population aging, lifestyle variation and other factors, the morbidity and mortality of cancer continue to rise in China, resulting in a serious public health problem threating people's health. In response to this increasingly serious problem, tremendous efforts have been made in China, including the development of a series of policies and measures for specific needs of cancer prevention and treatment. Currently, China's strategy to reduce the disease burden of cancer mainly focuses on primary and secondary prevention. This paper summarizes the current status of disease burden of cancer and the performance of three-grade cancer prevention in China.

Objective:To understand the epidemiological characteristics and spatiotemporal distribution of viral encephalitis in children and adolescents in Henan Province from 2012 to 2023.Methods:The information about viral encephalitis cases from October 1, 2012 to July 26, 2023 were collected from Zhengzhou Children's Hospital (National Children's Regional Medical Center),Henan Provincial Children's Hospital for the analyses on temporal distribution the cases, the severe illness rate, age distribution, pathogen type and imaging findings of the cases.Results:A total of 6 276 cases of viral encephalitis were included in this study after excluding cases with incomplete information. The cases mainly originated from Zhengzhou (38.96%), followed by Zhoukou (9.93%), Xuchang (8.68%), Zhumadian (7.90%) and Pingdingshan (7.39%). The cases in boys accounted for 62.13% and the cases in girls accounted for 37.87%. Most cases (72.45%) occurred in age group 7-13 years. The overall rate of severe illness cases was 4.51% from 2012 to 2023. There were significant differences in severe illness cases among different areas and years ( χ2=5.33, P=0.021; χ2=48.14, P<0.001). Enteroviruses were mainly detected (31.57%), in which Coxsackie virus was predominant (58.37%). Imaging findings showed that cerebral hemisphere damage was most common in children and adolescents with viral encephalitis (54.93%). Conclusions:From 2012 to 2023, more cases of viral encephalitis occurred in boys in Henan. Children and adolescents aged 7-13 years were the main affected group. The prevention of enteroviruses infection, especially Coxsackie virus, needs to be strengthened. Special attention should be paid to the prevention of cerebral hemisphere damage after viral encephalitis diagnosis.

Objective:To explore the treatment methods and clinical efficacy of penetrating high-voltage electric burns to thoracoabdominal wall.Methods:This study was a retrospective observational study. From March 2020 to March 2023, six male patients with penetrating high-voltage electric burns to thoracoabdominal wall who met the inclusion criteria were hospitalized at West China Hospital of Sichuan University, aged 42 to 57 years. Among them, one patient had thoracoabdominal wall defects with perforation and necrosis of the gastric wall and diaphragm, two patients had thoracoabdominal wall defects with perforation and necrosis of the gastric wall alone, and three patients had abdominal wall defects with perforation and necrosis of the small intestine. In the emergency department, aesthetic plastic and burn surgery, general surgery, and/or thoracic surgery doctors jointly formulated an emergency surgery plan for the patients. Three patients with perforation and necrosis of the gastric wall underwent subtotal gastrectomy and anastomosis. One of them, who also had diaphragmatic perforation and necrosis, underwent resection of the necrotic diaphragm and repair. The other three patients with perforation and necrosis of the small intestine underwent resection and anastomosis of the necrotic intestinal segment. After debriding the thoracoabdominal wall wounds conservatively in stage Ⅰ and repairing the abdominal wall defects with greater omentum coverage, the thoracoabdominal wall wounds were treated with vacuum sealing drainage (VSD). Seven days later, a stage Ⅱ thorough debridement of the thoracoabdominal wall wounds was performed, and bovine acellular dermal matrix was transplanted onto the surface of the greater omentum and the surrounding skin wounds without skin coverage, and the VSD treatment of the thoracoabdominal wall wounds was continued. After 7 days, the VSD treatment was stopped, and after the fresh granulation tissue well developed in the wounds, a stage Ⅲ transplantation of meshed split-thickness skin graft from the lateral femur was performed to close the thoracoabdominal wall wounds, followed by continuing VSD treatment for another 7 days. Data were recorded including the status of patients' enteral nutrition recovery and occurrence of complications such as abdominal infection after stage Ⅰ surgery, the time from starting moist dressing change to the wound being ready for skin grafting after stage Ⅱ surgery, the microbiological culture of wound exudate samples and conditions of wound surrounding areas from starting moist dressing change to wound healing after stage Ⅱ surgery, skin graft survival, and wound healing time. Follow-up was carried out to observe the occurrence of gastrointestinal symptoms, abdominal wall hernia, scars, and functional disorders, etc.Results:All six patients resumed enteral nutrition on day 2 to 4 after stage Ⅰ surgery, with no occurrence of intestinal obstruction, anastomotic leakage, or abdominal infection. The time from starting moist dressing change to the wound being ready for skin grafting after stage Ⅱ surgery was 8 (6, 12) days. During the period from starting moist dressing change to wound healing after stage Ⅱ surgery, microbiological culture of wound exudate sample showed Enterobacter cloacae in one patient and Pseudomonas aeruginosa in another patient, while the remaining four patients had negative cultures; no patient developed wound edge inflammation. All grafted skin survived, and the wound healing time was 38 (30, 46) days. During follow-up from 12 to 36 months after stage Ⅲ surgery, patients had no intractable constipation or intestinal obstruction symptoms, no obvious local tissue herniation requiring surgical treatment, no scar ulceration, and no functional disorders. Conclusions:For penetrating high-voltage electric burns to thoracoabdominal wall, a sequential three-stage surgical treatment plan under a multidisciplinary team collaboration model can effectively protect organ function and control wound infection. Long-term follow-up shows good gastrointestinal function and thoracoabdominal wall appearance. This method is highly reproducible and effective, which is suitable for clinical promotion and use.

Objective To explore the effect of ligustrazine on the learning and memory function of rats with aluminum-induced cognitive impairment and its mechanism.Methods Sixty male Wistar rats were divided into a blank control group,a model group,a low-dose ligustrazine group,a high-dose ligustrazine group,and a piracetam group using a random number table method,with 12 rats in each group.The rats in the blank control group were not subjected to any treatment;the rats in the model group,low-dose ligustrazine group,high-dose ligustrazine group,and piracetam group were first prepared with aluminum toxicity models by daily gavage of 100 mg·kg-1 AlCl3 solution.After successful modeling,the rats in the piracetam group were intragastrically administered with piracetam at a dose of 400 mg·kg-1,while rats in the low-dose and high-dose ligustrazine groups were intragastrically administered with 100 and 200 mg·kg-1 ligustrazine,respectively;the rats in the blank control group and the model group were intragastrically administered with the same volume of physiological saline.All rats in the five groups received intragas tric administration once a day for 30 consecutive days.After 30 days of intervention,the Morris water maze test was used to evaluate the learning and memory function of rats in the five groups.After completing the water maze experiment,rats in the five groups were anesthetized with 200 g·L-1 chloral hydrate,and their brain tissues were quickly removed after decapitation.Immunohistochemical staining was used to observe the expression of calcium voltage-gated channel subunit alpha 1E(CACNA1E),calmodulin(CALM),and brain-derived neurotrophic factor(BDNF)in the hippocampal CA1 region of rats in the five groups;Western blot was used to detect the relative expression levels of CACNA1E,CALM,and BDNF proteins in the hippocampal CA1 region of rats in the five groups;and real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression levels of CACNA1E,CALM,and BDNF mRNA in the hippocampal CA1 region of rats in the five groups.Results On the 1st day of the Morris water maze test,the latent periods of rats in the model group,piracetam group,low-dose ligustrazine group,and high-dose ligustrazine group were significantly higher than those in the blank control group(P＜0.05);there was no statistically significant difference in the latent periods of rats among the piracetam group,low-dose ligustrazine group,high-dose ligustrazine group,and model group(P＞0.05).On the 3rd day of the Morris water maze test,the latent periods of rats in the model group,piracetam group,low-dose ligustrazine group,and high-dose ligustrazine group were significantly higher than those in the blank control group(P＜0.05);the latent periods of rats in the piracetam group and high-dose ligustrazine group were significantly lower than those in the model group and low-dose ligustrazine group(P＜0.05);there was no statistically significant difference in the latent periods of rats between the low-dose ligustrazine group and the model group(P＞0.05).On the 5th day of the Morris water maze test,the latent periods of rats in the model group,piracetam group,low-dose ligustrazine group,and high-dose ligustrazine group were significantly higher than those in the blank control group(P＜0.05);the latent periods of rats in the piracetam group and high-dose ligustrazine group were significantly lower than those in the model group and low-dose ligustrazine group(P＜0.05);there was no statistically significant difference in the latent periods of rats between the low-dose ligustrazine group and the model group(P＞0.05).On the 3rd and 5th days of the Morris water maze test,there was no statistically significant difference in the latent periods of rats between the piracetam group and the high-dose ligustrazine group(P＞0.05).The times of rats crossing platform in the model group,piracetam group,low-dose ligustrazine group,and high-dose ligustrazine group were significantly lower than those in the blank control group,and the times of rats crossing platform in the piracetam group and high-dose ligustrazine group were significantly higher than those in the model group and low-dose ligustrazine group(P＜0.05);there was no statistically significant difference in the times of rats crossing platform between the low-dose ligustrazine group and the model group(P＞0.05);there was no statistically significant difference in the times of rats crossing platform between the piracetam group and the high-dose ligustrazine group(P＞0.05).Under the microscope,brown CACNA1E,CALM,and BDNF positive cells could be observed in the hippocampal CA1 region.The expressions of CACNA1E,CALM,and BDNF proteins in the hippocampal CA1 region of rats in the model group,piracetam group,low-dose ligustrazine group,and high-dose ligustrazine group were significantly lower than those in the blank control group,and the expressions of CACNA1E,CALM,and BDNF proteins in the hippocampal CAl region of rats in the piracetam group and high-dose ligustrazine group were significantly higher than those in the model group and low-dose ligustrazine group(P＜0.05);there was no statistically significant difference in the expressions of CACNA1E,CALM,and BDNF proteins in the hippocampal CAI region of rats between the low-dose ligustrazine group and the model group(P＞0.05);there was no statistically significant difference in the expressions of CACNA1E,CALM,and BDNF proteins in the hippocampal CA1 region of rats between the piracetam group and the high-dose ligustrazine group(P＞0.05).The relative expression levels of CACNA1E,CALM,and BDNF proteins and mRNAs in the hippocampal CA1 region of rats in the model group,piracetam group,low-dose ligustrazine group,and high-dose ligustrazine group were significantly lower than those in the blank control group(P＜0.05);the relative expression levels of CACNA1E,CALM,and BDNF proteins and mRNAs in the hippocampal CA1 region of rats in the piracetam group and high-dose ligustrazine group were significantly higher than those in the model group and low-dose ligustrazine group(P＜0.05);there was no statistically significant difference in the relative expression levels of CACNA1E,CALM,and BDNF proteins and mRNAs in the hippocampal CA1 region of rats between the low-dose ligustrazine group and the model group(P＞0.05);there was no statistically significant difference in the relative expression levels of CACNA1E,CALM,and BDNF proteins and mRNAs in the hippocampal CA1 region of rats between the piracetam group and the high-dose ligustrazine group(P＞0.05).Conclusion Ligustrazine has significant protective effects on aluminum-induced cognitive impairment in rats and can greatly enhance the learning and memory function of rats.The mechanism may be related to the up-regulation of CANA1E,CALM and BDNF expression in the brain induced by ligustrazine.

Objective:To seek the optimal melanin-removal method for hematoxylin and eosin (HE) staining, immunohistochemistry and molecular detection.Methods:Thirty-eight paraffin tissue samples of malignant melanoma diagnosed at the Fujian Cancer Hospital, Fuzhou, China between January 2018 and March 2022 were collected and used to make a tissue microarray. Melanin in these cases was removed using warm hydrogen peroxide, double oxidation depigmentation, modified potassium permanganate-oxalic acid or trichloroisocyanuric acid, followed by HE staining. The cases were divided into two cohorts: one was subject to the one of the above four methods to remove melanin first, followed by immunohistochemistry (SOX-10, Ki-67, HMB45 and Melan A), while the other was subject to immunohistochemical staining first and then a melanin removal. Following that, seventeen melanin-rich paraffin tissue samples were collected and depigmented using the methods described above. DNA extraction was then done, followed by assessments of DNA content and quality. Moreover, the completeness of melanin removal, the effect on HE and immunohistochemical staining, and the quality of DNA were compared between the depigmented methods.Results:Regarding the effectiveness of melanin removal, the modified potassium permanganate-oxalic acid and the warm hydrogen peroxide methods were the most effective, and both showed residual melanin in only 5.26% (2/38) of the cases. The trichloroisocyanuric acid method showed residual melanin in 10.53% (4/38) of the cases. The worst was the double oxidation depigmentation method, which showed pigment residue in 15.79% (6/38) of the cases. For HE staining, the percentage of good staining with the warm hydrogen peroxide method was 92.11%, higher than the other three methods. For immunohistochemical staining, the mean staining scores of immunohistochemistry first followed by melanin removal with modified potassium permanganate-oxalic acid, double oxidation and trichloroisocyanuric acid were 20.84, 26.63 and 35.02, respectively. These immunohistochemical staining scores were higher than those of melanin removal first followed by immunohistochemistry (8.70, 15.41 and 21.22, respectively). The mean staining score of melanin removal by warm hydrogen peroxide method followed by immunohistochemistry was 33.57, superior to that of immunohistochemistry followed by the melanin removal (19.96). Moreover, the staining scores of HMB45, MelanA and Ki-67 with immunohistochemical staining followed by trichloroisocyanuric acid method were 36.45, 33.79, and 36.24, respectively, while the staining score of SOX10 with melanin removal by warm hydrogen peroxide followed by immunohistochemistry was 34.39. The DNA was significantly degraded by modified potassium permanganate-oxalic acid, double oxidation depigmentation and trichloroisocyanuric acid, whereas the mean concentration of DNA extracted after melanin removal by hydrogen peroxide method was 59.59 μg/L, substantially higher than that of DNA extracted without melanin removal (30.3 μg/L, P=0.001). The A260/ A280 of DNA extracted after melanin removal by hydrogen peroxide was between 1.8 and 2.0 in all cases, and the A260/ A230 was above 2.0 in sixteen cases, suggesting high purity of DNA. However, the DNA extracted without removing the melanin showed poor purity, with A260/ A280 below 1.8 in eight cases and A260/ A230 below 2.0 in sixteen cases. Conclusions:Warm hydrogen peroxide showed the least melanin residue, superior HE staining and a minimal effect on DNA purity/quality compared to the other three methods. It thus appears most suitable for PCR, NGS and other molecular detection. Melanin removal with trichloroisocyanuric acid after immunohistochemical staining has the least melanin residual, and thus could be the most convenient and efficient. However, it is noted that the efficacy of the same depigmentation method varies with different antibodies. Therefore, the optimal depigmentation method should be selected based on the specific markers of interest.

Objective:To investigate the efficacy of different types of neoadjuvant therapy for esophageal cancer.Methods:The clinical data of 542 patients with esophageal squamous cell carcinoma (ESCC) who received neoadjuvant therapy in Anyang Tumor Hospital of Science and Technology from January 2015 to May 2022 were retrospectively analyzed. These patients, consisting of 198 females and 344 males, with 289 cases aging ≤ 65 and 253 cases aging >65, were divided into a neoadjuvant chemoradiotherapy (NCRT) group (137 cases), a neoadjuvant chemotherapy (NCT) group (241 cases), and a neoadjuvant immunotherapy plus chemotherapy (NICT) group (164 cases). In this study, primary endpoints included major pathological response (MPR) and pathologic complete response (pCR) rates, and secondary endpoints comprised overall survival (OS), progression-free survival (PFS), and safety. Survival analysis was performed using the Kaplan-Meier method, and inter-group comparisons were made using the Log-rank test. Furthermore, prognostic factors were analyzed based on the Cox proportional hazards regression model.Results:The NCRT, NCT, and NICT groups exhibited MPR and pCR rates of 66.4% (91/137) and 35.3% (85/241), 63.4% (104/164) and 35.8% (49/137), and 6.6% (16/241) and 31.1% (51/164), respectively ( χ2=1.67, P < 0.001). These groups displayed 1-, 2-, and 3-year OS rates of 89.8%, 85.9%, and 91.9%; 82.3%, 71.4%, and 81.5%; and 72.3%, 61.4%, and 77.8%, respectively, with significant differences ( χ2=9.20, P < 0.01). Furthermore, they exhibited 1-, 2-, and 3-year PFS rates of 81.5%, 75.9%, and 80.1%; 67.9%, 61.0%, and 65.5%; and 66.6%, 53.5%, and 65.3%, respectively, with significant differences ( χ2=4.62, P < 0.05). Multivariate analysis showed that therapeutic modality, T stage, and N stage were independent prognostic factors for OS ( P < 0.05). Additionally, there was no difference in adverse reactions and postoperative complications among the three groups. Conclusions:Compared to NCT, NICT and NCRT feature higher pCR and MPR rates, along with more survival benefits. Therefore, neoadjuvant immunotherapy has the potential to serve as a preoperative therapeutic modality for esophageal cancer, yet large-scale randomized controlled trials are still required for confirmation.