The mechanism of Tibetan medicine Zuotangkaca pills (ZTKCW) for the treatment of hypertension was explored by network pharmacology and in vivo experiments. 68 active ingredients of ZTKCW and 518 drug-disease targets were screened by network pharmacology. Eight core components of ZTKCW (vasicolinone, luteolin, (–)-isocorypalmine, esculetin, liquiritigenin, etc.) and eight key targets (AKT1, TNF, IL6, and STAT3, etc.) were screened by network topology analysis. KEGG enrichment analysis showed that the core targets were mainly enriched in lipids and atherosclerosis, JAK/STAT, and inflammation-related pathways. An in vivo experiment was conducted using spontaneously hypertensive rats (SHR), which were gavaged with ZTKCW at doses of 0.41, 0.82, and 1.64 g/kg for 12 weeks, respectively. The results showed that ZTKCW at a dose of 1.64 g/kg significantly reduced both systolic and diastolic pressure in SHR rats and decreased the phosphorylation levels of AKT1, PI3K, STAT3, and JAK2 in the thoracic aorta and heart tissues. This study demonstrates that ZTKCW may exert its antihypertensive effects through PI3K/AKT and JAK2/STAT3 pathways, providing some insights and a theoretical basis for the use of ZTKCW in hypertension.

Objective:To analyze the acupoint selection law in treating insomnia with warming-needle moxibustion using data mining technology.Methods:The clinical research literature on warming-needle moxibustion in treating insomnia was retrieved from the databases of CNKI, VIP, Wanfang Data, China Medical Journal Full-text Database, SinoMed, PubMed from the establishment of the databases to June 30, 2023. Acupuncture prescriptions were extracted and a database was established. Descriptive analysis was carried out on the intervention measures, the number of moxibustion numbers, the total effective rate, and the relevant information of acupoint selection. Clustering analysis and association rule analysis by using SPSS Statistics 26.0, SPSS Modeler 18.0 were performed on the high-frequency acupoints used in body acupuncture and warming acupuncture, respectively.Results:A total of 67 articles about warming-needle moxibustion for insomnia were included. The main intervention was single warming-needle moxibustion, followed by combined auricular acupuncture. Moxibustion with moxa sticks of 1 to 2 cm in length was often used. The total effective rate was mostly between 92% and 94%. A total of 85 prescriptions were obtained, involving 55 acupoints (569 times in frequency), among which Baihui (DU 20) (55 times), Shenmen (HT 7) (54 times) and Sanyinjiao (SP 6) (45 times) were widely used. The selected acupoints were mainly distributed on the head, face, back and waist, and the affiliated meridians were mainly bladder meridian, Governor Vessel and heart meridian. The specific acupoints were mainly crossing points, followed by back-shu acupoints. Clustering analysis of high-frequency acupoints obtained three effective groups, including "Xinshu (BL 15)- Pishu (BL 20) - Shenshu (BL 23) - Ganshu (BL 18)", etc. Baihui (DU 20) and Shenmen (HT 7) were the most relevant acupoint combinations. A total of 44 acupoints (frequency 272 times) were involved in the selection of warming acupuncture, among which Sanyinjiao (SP 6) (32 times), Xinshu (BL 15) (23 times) and Pishu (BL 20) (23 times) were more frequently used. Clustering analysis of high-frequency acupoints obtained three effective groups, including "Xinshu (BL 15)- Pishu (BL 20) - Shenshu (BL 23) - Ganshu (BL 18)", etc. Pishu (BL 20) and Xinshu (BL 15) were the most relevant acupoint combinations.Conclusions:Warming-needle moxibustion for the treatment of insomnia focuses on the application of acupoints on the bladder meridian on the back and waist, cooperates with the selection of acupoints on the head and face, and attaches importance to the application of intersection points in specific acupoints. The selection of acupoints by warming-needle moxibustion is mainly based on the back-shu acupoints of zang-fu organs, which plays the role of balancing yin and yang, regulating zang-fu organs, tonifying deficiency and tranquilizing mind to help sleep.

Objective:To observe any effect of dry needling of myofascial trigger points on substance P and synaptophysin expression in the spinal dorsal horn.Methods:Sixty-four Sprague-Dawley rats were randomly divided into a control group ( n=16) and a model group ( n=48). Myofascial trigger points were induced in the model group by a blunt strike and eccentric running. That group was then randomly divided into a no-treatment group ( n=15), a massage group ( n=16), and a dry needling group (16 rats). The rats in the two treatment groups received 4 weeks of dry needling or Chinese massage. Pressure pain thresholds were recorded before the experiment and after the 4 weeks. The content of substance P and synaptophysin in the spinal dorsal horn were measured using immunoblotting and immunohistochemistry. Results:After the treatment 14 rats (93%) in the model group had trigger points, significantly higher than the 8 rats (50%) in the massage group and the 7 rats (44%) in the dry needling group. After treatment, the average pressure pain thresholds of the no-treatment and massage groups was significantly lower than the control group′s average, while the difference between the dry needling group and the control group was not significant. The average pressure pain threshold had improved significantly in the no-treatment group, the massage group and the dry needling group, and the averages of the massage group and the dry needling group were significantly higher than that of the no-treatment group. The level of substance P was significantly higher in the no-treatment group than in the other three groups and the ratio of substance P to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH)was significantly higher. The substance P: GAPDH ratio of the massage group was significantly higher than that of the control group.Conclusions:Dry needling and massage are effective in relieving myofascial pain, at least in rats. Both can reduce the content of substance P in the spinal dorsal horn.

Objective To investigate the effect of high fat on the expression of high-mobility group box 1 (HMGB1),α-smooth muscle actin (α-SMA),and matrix metalloproteinase-2 (MMP-2) in rat hepatic stellate cells (HSC-T6 cells).Methods HSC-T6 cells were cultured in vitro and treated with palmitic acid (PA) at a concentration of 300 μmol/L for 24 hours,and the HSC-T6 cells in the control group were treated with bovine serum albumin (BSA) of the same volume for 24 hours;Westem blot was used to measure the expression of α-SMA and MMP-2.The HSC-T6 cells in the dose-effect group were treated with PA at concentrations of 0,100,200,300,500,and 1000 μmol/L for 24 hours;the HSC-T6 cells in the time-effect group were treated with PA for 0,4,8,12,24,and 48 hours;in the control group,PA was replaced by BSA of the same volume;Western blot was used to measure the expression of HMGB 1,α-SMA,and MMP-2.The HSC-T6 cells were treated with recombinant HMGB1 (rHMGB1) at concentrations of 0,50,100,200,and 500 ng/ml for 48 hours,and in the control group,PA was replaced by phosphate buffer of the same volume.Western blot was used to measure the expression of α-SMA and MMP-2.The HSC-T6 cells in the PA group were treated with PA at a concentration of 300 μmol/L for 24 hours;the HSC-T6 cells in the PA+HMGB1-siRNA group were treated with PA at a concentration of 300 μmol/ L for 24 hours after HMGB1 in HSC-T6 cells was down-regulated;in the blank control group,PA was replaced by BSA of the same volume.Western blot was used to measure the expression ofHMGB1,α-SMA,and MMP-2.One-way analysis of variance was applied for continuous data,and the t-test was applied for comparison between two groups.Results (1) The expression of α-SMA and MMP-2 increased significantly after HSC-T6 cells were treated with PA at a concentration of 300 μmol/L for 24 hours (P ＜ 0.05).(2) Compared with the HSC-T6 cells in the untreated group (0 μmol/L),the HSC-T6 cells treated with different concentrations of PA showed significant increases in the expression of HMGB1 and MMP-2 (P ＜ 0.01),as well as a significant increase in the expression of α-SMA at concentrations of 200,300,500,and 1000 μmol/L (P ＜ 0.01);the HSC-T6 cells treated with PA at a concentration of 300 μmol/L for different periods of time showed varying degrees of increase in the expression of HMGB1,α-SMA,and MMP-2,with significant increases at 16,24,and 48 hours (P ＜ 0.01).(3) Compared with the HSC-T6 cells in the untreated group (0 ng/ml),the HSC-T6 cells treated with rHMGB 1 at concentrations of 100,200,and 500 ng/ml for 48 hours showed significant increases in the expression of α-SMA and MMP-2 (P ＜ 0.05).(4) Compared with the HSC-T6 cells in the PA group,the HSC-T6 cells treated with PA+HMGB1-siRNA for 24 hours showed significant reductions in the expression of HMGB 1,α-SMA,and MMP-2 (P ＜ 0.05).Conclusions High fat can increase the expression of α-SMA and MMP-2 through up-regulating the expression of HMGB 1 in HSC-T6,and thus lead to the development of liver fibrosis.