Objective:To explore the effect of necroptosis inhibitor necrosulfonamide (NSA) on traumatic brain injury (TBI) mouse model and BV2 cell pyroptosis model and their mechanisms.Methods:(1) In vivo experiments: 50 mice were randomly divided into sham-operated group, TBI group, TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group, and TBI+10 mg/kg NSA group, with 10 mice in each group. TBI model was established using a modified Feeney's weight-drop method; 4 h after modeling, 90% corn oil, 1 mg/kg NSA, 5 mg/kg NSA, or 10 mg/kg NSA was administered into the mice, respectively. Mice in the sham-operated group only had circular bone window opened without being subjected to impact. At 48 hours after modeling, neurological function was evaluated by modified neurological function score (mNSS), serum lactate dehydrogenase (LDH) content was detected by LDH detection kit, contents of interleukin (IL)-18, IL-1β and tumor necrosis factor-α (TNF-α) in the brain tissues were detected by enzyme-linked immunosorbent assay (ELISA), and expressions and localizations of ionized calcium binding adaptor molecule 1 (IBA-1), cysteinyl aspartate specific proteinase-1 (Caspase-1) p20 and gasdermin D (GSDMD) in the injured parietal cortex were detected by double immunofluorescent staining. (2) In vitro experiments: BV2 cells were divided into control group, lipopolysaccharide (LPS)+adenosine triphosphate (ATP)+dimethyl sulfoxide (DMSO) group, LPS+ATP+5 μmol/L NSA group, LPS+ATP+10 μmol/L NSA group, and LPS+ATP+15 μmol/L NSA group. Cells in the latter 4 groups were induced by LPS+ATP to establish BV2 cell pyroptosis model, and incubated with 2 μL DMSO, 5 μmol/L NSA, 10 μmol/L NSA, and 15 μmol/L NSA for 1 hour, respectively; cells in the control group were cultured conventionally. Contents of LDH, IL-1β, IL-18, and TNF-α in the cell culture supernatant were detected by ELISA; pyroptosis was detected by calcein acetoxymethyl ester (CAM)/propidium iodide (PI) double staining; protein expressions of nucleotide binding domain-like receptor protein 3 (NLRP3), Caspase-1 p20, GSDMD, and N-terminal fragment of GSDMD (GSDMD-N) were detected by Western blotting. Results:(1) Compared with the TBI group, the TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group and TBI+10 mg/kg NSA group had decreased mNSS score and serum LDH content, decreased IL-1β and IL-18 contents in the brain tissues and number of Caspase-1 p20 + cells in the injured parietal cortex, successively, with significant differences ( P<0.05). Compared with the TBI group ([287.80±12.26] cells/mm 2), the TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group, and TBI+10 mg/kg NSA group had decreased number of Iba-1 +GSDMD + cells in the injured parietal cortex ([213.70±11.87] cells/mm 2, [205.30±9.15] cells/mm 2, [131.70±13.69] cells/mm 2),successively, with significant differences ( P<0.05). Compared with the TBI group, the TBI+5 mg/kg NSA group and TBI+10 mg/kg NSA group had significantly decreased number of Iba-1 + cells in the injured parietal cortex, and the TBI+10 mg/kg NSA group had significantly decreased TNF-α content in the brain tissues and number of GSDMD + cells in the injured parietal cortex ( P<0.05). Compared with the TBI group ([247.20±9.88] cells/mm 2), the TBI+10 mg/kg NSA group had significantly decreased number of Iba-1 +Caspase-1 p20 + cells in the injured parietal cortex ([181.70±9.37] cells/mm 2, P<0.05). (2) Compared with the LPS+ATP+DMSO group, the LPS+ATP+5 μmol/L NSA group, LPS+ATP+10 μmol/L NSA group, and LPS+ATP+15 μmol/L NSA group had decreased IL-18 content in the supernatant, successively, with significant differences ( P<0.05); and compared with the LPS+ATP+DMSO group, the LPS+ATP+10 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased contents of LDH, IL-1β, and TNF-α in the supernatant and ratio of PI +/CAM + cell counts ( P<0.05). Compared with the LPS+ATP+DMSO group (2.62±0.50), the LPS+ATP+10 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased Caspase-1 p20 protein expression (1.36±0.14, 1.32±0.07, P<0.05). Compared with the LPS+ATP+DMSO group (5.00±1.67), the LPS+ATP+5 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased GSDMD protein expression (1.42±0.26, 1.68±0.32, P<0.05). Compared with the LPS+ATP+DMSO group (2.28±0.24), the LPS+ATP+15 μmol/L NSA group had significantly decreased GSDMD-N protein expression (1.23±0.08, P<0.05). Conclusion:NSA can inhibit microglial pyroptosis after TBI by inhibiting the Caspase-1 p20/GSDMD pathway, thereby playing a neuroprotective role.

Objective To explore the distribution of circuit-qi indicators at birth in the patients with chronic atrophic gastritis(CAG)based on five-circuit and six-qitheory,and to summarize their congenital circuit-qi endowment features,so as to explore treatment strategies for CAG accordingly.Methods From January 2018 to December 2023,a total of 2 046 cases of patients with CAG admitted to Shenzhen Bao'an Traditional Chinese Medicine Hospital Affiliated to Guangzhou University of Chinese Medicine(i.e.,Shenzhen Bao'an Traditional Chinese Medicine Hospital)were selected,and the distribution of the five-circuit and six-qiindicators at birth in the patients was analyzed by using the SPSS 24.0 statistical software.The features of the congenital circuit-qi endowment were summarized,and then the corresponding treatment strategies were put forward.Results The CAG patients had the highest proportions of circuit-qiendowment features at birth,showing as the yearly circuit being deficiency fire circuit,the dominant circuit being metal circuit,the dominant qibeing yangmingdryness-gold,the guest qibeing taiyangcold-water,and sitianbeing shaoyangminister fire and zaiquanbeing j ueyinwind-wood,and the differences in the distribution of the dominant circuit,dominant qi,and guest qiwere statistically significant(P<0.05 or P<0.01),but differences in the distribution of the yearly circuit and sitian/zaiquanwere not statistically significant(P>0.05).The analysis of the combined circuit-qifeatures at birth showed that the proportion of patients with CAG born in the year of celestial restriction,mildly-rebellious year and disharmony year was the highest,and the proportion of those born in the convergent year and iso-convergent year was the lowest,with statistically significant differences(P<0.05).Conclusion The congenital circuit-qiendowment of patients with CAG is characterized by liver depression and qistagnation,dryness and coldness.Therefore,the treatment for CAG should be from the perspective of the liver,with the method of soothing liver and harmonizing stomach,supplemented by nourishing yin to moisten dryness,and protecting yang qiof the spleen and stomach.The combined use of acup-moxibustion and Chinese medicines is recommended.

ObjectiveTo observe the clinical efficacy of Shentong Zhuyutang combined with Dilongtang in the treatment of lumbar disc herniation （LDH） with Qi stagnation and blood stasis syndrome， and its effect on nucleus pulposus reabsorption and immune-inflammatory factors， exploring its therapeutic mechanism from the perspective of reabsorption. MethodsA total of 120 patients with LDH from the Fourth Clinical Medical College of Guangzhou University of Chinese Medicine， treated between June 2020 and January 2023， were randomly divided into the control group （52 cases， with 8 dropouts） and the observation group （49 cases， with 11 dropouts） according to a random number table. The control group received routine treatment， while the observation group was treated with Shentong Zhuyutang combined with Dilongtang in addition to routine treatment. Visual Analogue Scale （VAS）， Oswestry Disability Index （ODI）， Japanese Orthopaedic Association （JOA） score， and traditional Chinese medicine （TCM） syndrome score were measured before treatment and after 3 courses of treatment. Venous blood samples were collected for the determination of serological indexes. MR examination was performed during the 6-month follow-up to calculate the absorption rate. ResultsAfter treatment， both groups showed significant reductions in VAS， ODI， TCM syndrome score， serum tumor necrosis factor （TNF）-α， matrix metalloproteinase （MMP）-9， and vascular endothelial growth factor （VEGF） levels， and a significant increase in JOA score compared with pre-treatment values （P<0.05）. Compared with the control group， the observation group showed significantly lower VAS， ODI， TCM syndrome score， serum TNF-α， MMP-9， and VEGF levels， and a significantly higher JOA score （P<0.05）. The proportion of nucleus pulposus reabsorption in the observation group was 57.14% （28/49）， significantly higher than 21.15% （11/52） in the control group （χ²=6.161， P<0.05）. ConclusionShentong Zhuyutang combined with Dilongtang can effectively relieve pain， improve lumbar function， and alleviate TCM clinical symptoms in LDH patients with Qi stagnation and blood stasis syndrome. Imaging findings suggest that the treatment promotes the reabsorption of nucleus pulposus protrusion， while laboratory testing shows reduced serum levels of TNF-α， MMP-9， and VEGF， which contribute to the rehabilitation of patients.

Objective:To explore the effect of necroptosis inhibitor necrosulfonamide (NSA) on traumatic brain injury (TBI) mouse model and BV2 cell pyroptosis model and their mechanisms.Methods:(1) In vivo experiments: 50 mice were randomly divided into sham-operated group, TBI group, TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group, and TBI+10 mg/kg NSA group, with 10 mice in each group. TBI model was established using a modified Feeney's weight-drop method; 4 h after modeling, 90% corn oil, 1 mg/kg NSA, 5 mg/kg NSA, or 10 mg/kg NSA was administered into the mice, respectively. Mice in the sham-operated group only had circular bone window opened without being subjected to impact. At 48 hours after modeling, neurological function was evaluated by modified neurological function score (mNSS), serum lactate dehydrogenase (LDH) content was detected by LDH detection kit, contents of interleukin (IL)-18, IL-1β and tumor necrosis factor-α (TNF-α) in the brain tissues were detected by enzyme-linked immunosorbent assay (ELISA), and expressions and localizations of ionized calcium binding adaptor molecule 1 (IBA-1), cysteinyl aspartate specific proteinase-1 (Caspase-1) p20 and gasdermin D (GSDMD) in the injured parietal cortex were detected by double immunofluorescent staining. (2) In vitro experiments: BV2 cells were divided into control group, lipopolysaccharide (LPS)+adenosine triphosphate (ATP)+dimethyl sulfoxide (DMSO) group, LPS+ATP+5 μmol/L NSA group, LPS+ATP+10 μmol/L NSA group, and LPS+ATP+15 μmol/L NSA group. Cells in the latter 4 groups were induced by LPS+ATP to establish BV2 cell pyroptosis model, and incubated with 2 μL DMSO, 5 μmol/L NSA, 10 μmol/L NSA, and 15 μmol/L NSA for 1 hour, respectively; cells in the control group were cultured conventionally. Contents of LDH, IL-1β, IL-18, and TNF-α in the cell culture supernatant were detected by ELISA; pyroptosis was detected by calcein acetoxymethyl ester (CAM)/propidium iodide (PI) double staining; protein expressions of nucleotide binding domain-like receptor protein 3 (NLRP3), Caspase-1 p20, GSDMD, and N-terminal fragment of GSDMD (GSDMD-N) were detected by Western blotting. Results:(1) Compared with the TBI group, the TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group and TBI+10 mg/kg NSA group had decreased mNSS score and serum LDH content, decreased IL-1β and IL-18 contents in the brain tissues and number of Caspase-1 p20 + cells in the injured parietal cortex, successively, with significant differences ( P<0.05). Compared with the TBI group ([287.80±12.26] cells/mm 2), the TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group, and TBI+10 mg/kg NSA group had decreased number of Iba-1 +GSDMD + cells in the injured parietal cortex ([213.70±11.87] cells/mm 2, [205.30±9.15] cells/mm 2, [131.70±13.69] cells/mm 2),successively, with significant differences ( P<0.05). Compared with the TBI group, the TBI+5 mg/kg NSA group and TBI+10 mg/kg NSA group had significantly decreased number of Iba-1 + cells in the injured parietal cortex, and the TBI+10 mg/kg NSA group had significantly decreased TNF-α content in the brain tissues and number of GSDMD + cells in the injured parietal cortex ( P<0.05). Compared with the TBI group ([247.20±9.88] cells/mm 2), the TBI+10 mg/kg NSA group had significantly decreased number of Iba-1 +Caspase-1 p20 + cells in the injured parietal cortex ([181.70±9.37] cells/mm 2, P<0.05). (2) Compared with the LPS+ATP+DMSO group, the LPS+ATP+5 μmol/L NSA group, LPS+ATP+10 μmol/L NSA group, and LPS+ATP+15 μmol/L NSA group had decreased IL-18 content in the supernatant, successively, with significant differences ( P<0.05); and compared with the LPS+ATP+DMSO group, the LPS+ATP+10 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased contents of LDH, IL-1β, and TNF-α in the supernatant and ratio of PI +/CAM + cell counts ( P<0.05). Compared with the LPS+ATP+DMSO group (2.62±0.50), the LPS+ATP+10 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased Caspase-1 p20 protein expression (1.36±0.14, 1.32±0.07, P<0.05). Compared with the LPS+ATP+DMSO group (5.00±1.67), the LPS+ATP+5 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased GSDMD protein expression (1.42±0.26, 1.68±0.32, P<0.05). Compared with the LPS+ATP+DMSO group (2.28±0.24), the LPS+ATP+15 μmol/L NSA group had significantly decreased GSDMD-N protein expression (1.23±0.08, P<0.05). Conclusion:NSA can inhibit microglial pyroptosis after TBI by inhibiting the Caspase-1 p20/GSDMD pathway, thereby playing a neuroprotective role.

Objective To investigate the role of TLR4/NF-κB signal pathway in pathogenesis of brain inju-ry during deep hypothermia circulatory arrest(DHCA). Methods BV2 microglia cells were subjected to oxygen-glucose deprivation/reoxygenation(OGD/R),in vitro model for DHCA. The BV2 were randomly divided into the control group(C group)and the experimental group(O group). BV2 viability was determined by CCK-8 assay. TLR4 and its downstream signaling molecules ,MyD88 and phosphorylated NF-κB (p-p65) expressions were detected by Western blotting. TLR4 mRNA expression in BV2 microglial cells were determined by RT-PCR. Level of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in culture medium was detected by ELASA. Results Compared with the group C,BV2 microglia cell viability in experiment group was obviously weaker(P<0.05). Expressions of TLR4,MyD88 and phosphorylated NF-κB(p-p65)from the experiment group increased remarkedly than those from the group C (P < 0.05). TLR4 mRNA level was higher significantly in the group O than in the group C (P < 0.01). Production of IL-6 and TNF-α in the group O were up-regulated apparently compared to the group C(P<0.01). Conclusion TLR4/NF-κB signaling pathway contributed to activation of BV2 microglia cells treated by OGD/Reoxygenation ,which was probably the exactly way that involved in pathogenesis of brain injury during deep hypothermia circulatory arrest.

Objective To describe a rhesus monkey model of temporal lobe epilepsy (TLE) established via repetitive unilateral intra-amygdala kainic acid (KA) injection and provide experimental basis for epileptogenic network and related research. Methods Eight male adult rhesus monkeys were randomly divided into KA injection group (n=6) and saline injection group (n=2). Brain stereotaxic technique, micro catheter implantation into the right amygdaloid nucleus, subcutaneous bladder connection, and continuous video-EEG monitoring were performed, and KA or saline injection into their right amygdala was achieved. Interictal epileptic discharges (IEDs), ictal discharges and behavioural performance between the two groups were compared right after injection and within 6 months of first discharge. Results Typical IEDs were recorded in the 6 monkeys from KA injection group after 2-4 times of KA injection, with focal spike waves discharges at the right temple area as manifestation; ictal discharges were recorded in 4 monkeys, with discharge patterns of discharges from the right temple area to the whole brain as manifestation, and during epileptic attack, these 4 monkeys suddenly stopped and dumbfounded without obvious limb seizures. Monkeys from the saline injection group showed no obvious abnormal behaviors. Conclusion Through a modified protocol of unilateral repetitive intra-amygdala KA injection, a rhesus monkey model with high similarity of behavioral and brain electrical features to TLE is developed.

Objective To investigate the status of soil fertility of Good Agricultural Practices ( GAP) base for Spatholobus suberectus Dunn. (SSD) in Pingyuan county of Guangdong province, thus to provide reference for GAP research and the subsequent fertilization for SSD. Methods The deep layer and superficial layer of GAP soil were collected for the physiochemical detection and nutrient assay. Compared with the classification standard of the second national general soil investigation, single base soil fertility index was diagnosed and the comprehensive soil fertility was evaluated with modified Nemoro Index. Results The soil pH value and the contents of exchangeable calcium and magnesium were unbalanced, and the contents of macroelements of nitrogen and phosphonium, microelements, and organic matter were low. Therefore, the measures for improving the base soil fertility should be as follows: ( 1) soil amendments of bentonite, gypsum and slaked lime should be used to adjust the soil pH value; ( 2) each plant should be given 10 kg of slaked organic fertilizer as base fertilizer; ( 3) in the process of nurturing, some special micro-fertilizer solution should be used to treat the cut slips, and 5 kg of urea should be used for every 667 meter square of land; ( 4) besides compound fertilizer, every 667 meter square of land should be fertilized with 15 kg of ammonium dihydrogen phosphate for the supplement of nitrogen and phosphorus, and slaked lime and magnesium carbonate should be used for the supplement of soil moderate-quantity elements after transplantation. Conclusion The comprehensive fertility of Pingyuan GAP base for Spatholobus suberectus Dunn. is at low level, and should be improved in combination with GAP requirements.

ObjectiveTo investigate the dietary intake of phytosterols in middle-aged and elderly residents in Guangzhou.Methods The dietary data were collected from 599 middle-aged and elderly residents (222 men and 377 women) recruited by stratified cluster random sampling in Guangzhou.All the subjects were surveyed using Food Frequency Questionnaire (FFQ).The dietary intake of phytosterols was estimated using Chinese data of phytosterol composition of a range of foods.ResultsThe dietary intake of total phytosterols was 336.36 ±142.88 mg/d,which included β-sitosterol 218.53 ± 95.20 mg/d,campesterol 48.33 ± 23.69 mg/d,stigmasterol 36.40 ± 14.38 mg/d,β-sitostanol 30.65 ± 13.62 mg/d,and campestanol 4.67 ±2.77 mg/d.Women had a significantly higher intake of phytosterols than men [ (345.45 ±141.06) mg/d vs.(320.93 ±144.95) mg/d,P=0.0425].Edible oil,vegetable,cereal,and fruit were the four major food sources of phytosterols,representing 37.2％,19.8％,18.5％,and 12.5％ of the total phytosterols intake respectively.Energy-adjusted intake of phytosterols was (42.94 ± 15.66) mg/1000 kJ,and women had a significantly higher intake than men [ (46.04 ±15.90) mg/1000 kJ vs.(37.69 ± 13.76) mg/1000 kJ,P =0.0000 ].ConclusionAmong the middle-aged and elderly residents in Guangzhou,women have higher phytosterols intake than men.

Objective To investigate the prevalence of metabolic syndrome(MS)and its components among people aged over 15 years in Guangxi Zhuang Autonomous Region and to compare the difference between Zhuang and Han populations.Methods Adopting cluster sampling,a survey of diabetes mellitus was conducted in Guangxi from 2003 to 2005.A total of 27 240 subjects aged over 15 years with complete data,including background information of each individual,blood pressure,lipid profile,plasma glucose,blood uric acid and fasting insulin were analyzed in this study.The prevalence of MS and its components were analysed in Han and Zhuang Chinese in Guangxi.The criteria of International Diabetes Federation(IDF)in 2005 and the China Diabetes Society(CDS)in 2004 were applied for diagnosis.Results(1)The crude prevalence rates of MS according to IDF definition were 13.15%in total,12.41%in male and 14.11%in female respectively.The age- standardized prevalence rates of MS(according to the population composition in China in 2000)were 7.66%in total,7.26%in male and 8.81%in female.The crude prevalence rates of MS according to CDS definition were 10.75%in total,13.45%in male and 7.28%in female respectively and the age-standar-dized prevalence rates of MS were 5.9%in total,7.21%in male and 4.31%in female.The prevalence of MS in total,male and female was increasing with age(P