Objective To investigate the neuroprotective effect and its mechanism of p38 mitogenactivated protein kinase inhibitor after subarachnoid hemorrhage (SAH).Methods Twenty-seven SPF-grade adult male SD rats were selected to induce a SAH model using the prechiasmal pool blood injection.Three dead rats were excluded.Twenty-four rats were randomly divided into four groups:sham operation group,SAH group,dimethyl sulfoxide (DMSO) group,and DMSO +p38 inhibitor group (n =6 in each group).Western blot was used to detect the expression levels of p38,phosphorylation p38,Parkinson's disease protein 7 (DJ-1),autophagy associated gene 5 (Atg5),autophagy adaptor protein p62,microtubule-associated protein Ⅰ Light Chain 3 (LC3-Ⅰ),microtubule-associated protein Ⅱ light chain 3 Ⅱ (LC3-Ⅱ),and the Garcia neurological function score was used to judge the nerve injury.PC12 cell oxygenated hemoglobin was used to induce an in vitro SAH model.They were completely randomly divided into four groups:sham operative group,SAH group,DMSO group,and DMSO + p38 inhibitor group.Fluorescent probe JC-1 was used to observe the changes of mitochondrial membrane potential.Results (1) There were significant differences in the expression of p38,phosphorylation-p38 and DJ-1 in rat brain tissue among the 4 groups (F values were 94.959,150.293 and 698.476,respectively,all P ＜ 0.01).There were significant differences in mitochondrial membrane potential in PC12 cells among the 4 groups (F value was 24.989,P ＜ 0.01).There were significant differences in the expression levels of autophagy related protein LC3-Ⅱ/LC3-Ⅰ ratio,Atg5 and p62 in rat brain tissue among the 4 groups (F values were 235.319,110.490 and 36.311,respectively,all P ＜ 0.01).There was significant difference in nerve function score among the 4 groups (F value was 25.550,P ＜ 0.01).(2) Compared with the sham operative group,the expression levels of p38,phosphorylation-p38 and DJ-1 were upregulated significantly after SAH (from 0.43 ±0.06,0.41 ±0.02 and 0.07 ±0.01 to 0.61 ± 0.08,0.79 ± 0.07 and 0.17 ± 0.03,respectively,all P ＜ 0.01),mitochondria membrane potential depolarization (from 8.29 ±0.28 to 9.23 ±0.42,P ＜0.01);upregulation of Atg5 expression and increase of LC3-Ⅱ/LC3-Ⅰratio (from 0.23 ± 0.04 and 0.25 ± 0.04 to 0.47 ± 0.04 and 0.46 ± 0.04,respectively,all P ＜ 0.01),down regulation of p62 expression (from 1.09 ± 0.14 to 0.54 ± 0.10,P ＜ 0.01);neurological score was decreased (from 17.5 ± 0.6 to 11.3 ± 2.7,P ＜ 0.01);p38 inhibitor was significantly down regulated the expression of phosphorylation-p38 after SAH (from 0.79 ± 0.07 to 0.47 ± 0.04,P ＜ 0.01),the expression of DJ-1 was up-regulated (from 0.17 ± 0.03 to 1.02 ± 0.06,P ＜ 0.01),mitochondrial membrane potential recovery (from 9.23 ±0.42 to 8.47 ±0.36,P ＜0.01),cell autophagy related protein LC3-Ⅱ/LC3-Ⅰ ratio and Atg5 were upregulated(from 0.46 ±0.04 and 0.47 ±0.04 to 0.77 ± 0.06 and 0.95 ± 0.12,all P ＜ 0.01),p62 expression returned to the levels of SAH group (from 0.57 ± 0.09,to 0.54 ± 0.10,P =0.650),and the neurological score was significantly improved (from 11.3 ± 2.7 to 15.5 ± 1.0,P ＜0.01).Conclusions After SAH,the p38 inhibitor downregulates the activity of2 phosphorylation p38.It may inhibit abnormal autophagy and maintain mitochondrial function by up-regulating the expression of DJ-1 protein,and then play a neuroprotective function.

Objective:To investigate the clinical efficacy of repetitive transcranial magnetic stimulation (rTMS) versus functional electrical stimulation (FES) in the treatment of post-stroke complex regional pain syndrome. Methods:The randomized controlled study included 60 patients with post-stroke complex regional pain syndrome who received treatment at the Wenzhou TCM Hospital of Zhejiang Chinese Medical University from July 2021 to February 2023. These patients were divided into an rTMS group ( n = 30) and an FES group ( n = 30) using a random number table method. Patients in the rTMS group were treated with rTMS, while those in the FES group were treated with FES. All treatments were performed once daily for 8 consecutive weeks. The clinical efficacy, total active movement score of the fingers, Fugl-Meyer assessment scale score, Activity of Daily Living score, modified Barthel Index score, shoulder joint range of motion, and safety were compared between the two groups. Results:The total response rate in the FES group was 86.7% (26/30), which was significantly higher than 83.3% (25/30) in the rTMS group ( Z = 0.09, P = 0.93). After treatment, there were no significant differences between the two groups in terms of total active movement score of the fingers and Fugl-Meyer assessment scale score ( P = 0.244, 0.262). No significant differences were found between the two groups in MBI score and ADL score (both P > 0.05). There was also no significant difference in shoulder joint range of motion between the two groups ( P > 0.05). Conclusion:Both rTMS and FES are highly effective for post-stroke complex regional pain syndrome. They can improve upper limb function, enhance daily living abilities, and remodel neurological functions of the brain.

AIM: To investigate t h e impacts of theaflavins (TFs) on neuronal apoptosis and blood-brain barrier (BBB) by regulating the calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2)/5 '-adenosine monophosphate-activated protein kinase (AMPK) signaling pathway. METHODS: Ninety rats were randomly separated into sham operation group, model group, low-dose TFs group (20 mg/kg TFs), high-dose TFs group (40 mg/kg TFs), and high-dose TFs + STO-609 group (40 mg/kg TFs + 10 ΜL CaMKK2 inhibitor-STO-609), positive control group (2 mg/kg nimodipine injection), with 15 rats in each group. A rat model of intracerebral hemorrhage was induced by collagenase type VII. The behavior of rats and the water content of brain tissue were detected; the serum of rats was isolated, and the levels of inflammatory factors-vascular cell adhesion molecule-1 (VCAM-1), tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule-1 (ICAM-1) were detected; brain tissue around the hematoma was collected to detect neuronal apoptosis, BBB permeability parameter-EB level, and expressions of p-CaMKK2/CaMKK2, p-AMPK/AMPK and apoptosis-related protein Bax. RESULTS: Compared with the sham operation group, the mNSS score, ICAM-1, TNF-α, VCAM-1, brain tissue water content, apoptosis rate, EB level and Bax protein expression in the model group were all increased, both pCaMKK2/CaMKK2 and p-AMPK/AMPK were decreased (P < 0.05); compared with the model group, the mNSS score, ICAM-1, TNF-α, VCAM-1, brain water content, apoptosis rate, EB level and Bax expression in the low- and high-dose TFs groups and the positive control group were all lower than those in the model group, both pCaMKK2/CaMKK2 and p-AMPK/AMPK were increased (P < 0.05); compared with the high-dose TFs group, the mNSS score, ICAM-1, TNF-α, VCAM-1, brain tissue water content, apoptosis rate, EB level and Bax expression were all increased in the high dose TFs + STO-609 group, both p-CaMKK2/CaMKK2 and p-AMPK/AMPK were decreased (P < 0.05). CONCLUSION: TFs can reduce neuronal apoptosis, inflammatory response, BBB permeability, and play a protective role in rats with cerebral hemorrhage injury. Its mechanism is related to the activation of CaMKK2/AMPK signaling pathway.

Objective:To explore the protective effect and mechanism of Tongfu Lifei decoction (TFL) on human monocytic leukemia cell THP-1 induced by lipopolysaccharide (LPS).Methods:① THP-1 cells were cultured in vitro, and incubated with 1 mg/L LPS for 18 hours to construct an in vitro THP-1 cell inflammation model. Other THP-1 cells were taken as blank control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) secreted by cells. ② THP-1 cells were divided into seven groups and treated with 0, 0.005, 0.01, 0.02, 0.04, 0.08, and 0.16 mL/mL TFL for 24 hours (added different dosages of TFL solution per milliliter of culture medium, with a crude drug content of 1 kg/L). The cell survival rate was detected using methyl thiazolyl tetrazolium (MTT) colorimetric method, and the intervention dosage of TFL for its non-toxic effect on THP-1 cells was screened. ③ Another THP-1 cells were divide into inflammatory model group and 0.01, 0.02, and 0.04 mL/mL TFL groups according to the intervention dosage of TFL screened by MTT colorimetry. After 24 hours of intervention, the levels of TNF-α and IL-6 secreted by cells were measured using ELISA. Western blotting was used to detect the expressions of programmed death-1/programmed death-ligand 1 (PD-1/PD-L1) signaling pathway proteins in cells. Real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expressions of microRNAs (miR-146a, miR-146b, miR-155) in cells. ④ The maximum non-toxic concentration of TFL (0.04 mL/mL) on the THP-1 cell was selected as the intervention dose. THP-1 cells were divided into inflammation model group, TFL group, TFL+miR-146a inhibitor group, TFL+miR-146b inhibitor group, and TFL+miR-155 inhibitor group. The inflammation model group was not given any drug intervention. The other inhibitor groups were added 100 nmol/L corresponding inhibitor. After 24 hours of intervention, the levels of TNF-α and IL-6 secreted by cells were measured using ELISA. Western blotting was used to detect the expressions of PD-1/PD-L1 signaling pathway proteins in cells. Results:① Compared with the blank control group, the levels of TNF-α and IL-6 secreted by cells in the inflammatory model group were significantly increased, indicating the successful construction of the THP-1 inflammatory cell model in vitro. ② 0-0.04 mL/mL TFL had no toxic effect on THP-1 cells. However, the survival rates of cells in the 0.08 mL/mL and 0.16 mL/mL TFL groups were significantly lower than those in the inflammation model group, indicating that TFL dosages exceeding 0.04 mL/mL had toxic effects on THP-1 cells. ③ Compared with the inflammation model group, 0.01 mL/mL TFL had no significant effect on the levels of TNF-α and IL-6 secreted by THP-1 cells, while intervention with 0.02 mL/mL and 0.04 mL/mL TFL significantly reduced the levels of TNF-α and IL-6 secreted by cells [TNF-α(ng/L): 95.89±8.55, 70.73±11.70 vs. 137.10±7.19, IL-6 (ng/L): 23.03±2.55, 16.58±1.72 vs. 32.60±2.55, all P < 0.01]. Compared with the inflammation model group, the expressions of PD-1/PD-L1 signaling pathway proteins in THP-1 cells in different dosages of TFL groups were significantly reduced, and showed a certain dosage dependence. The expressions of the pathway proteins in the 0.04 mL/mL TFL group were significantly lower than those in the inflammation model group [PD-1 protein (PD-1/β-actin): 0.28±0.04 vs. 1.00±0.10, PD-L1 protein (PD-L1/β-actin): 0.54±0.05 vs. 1.00±0.08, phosphoinositide 3-kinase (PI3K) protein (PI3K/β-actin): 0.28±0.03 vs. 1.00±0.08, phosphorylated protein kinase B (p-Akt) protein (p-Akt/Akt): 0.38±0.04 vs. 1.00±0.10, all P < 0.01]. Compared with the inflammation model group, the expression of miR-146a in THP-1 cells in the 0.01, 0.02, and 0.04 mL/mL TFL groups was significantly reduced (2 -ΔΔCt: 0.46±0.11, 0.31±0.13, 0.23±0.14 vs. 1.01±0.18, all P < 0.01), while there was no significant change in the expressions of miR-146b and miR-155. ④ Compared with the inflammation model group, the TFL group showed a significant decrease in the levels of TNF-α and IL-6 secreted by THP-1 cells. The miR-146a inhibitor could significantly reverse the inhibitory effect of TFL on inflammatory factors, and the difference was statistically significant as compared with the TFL group [TNF-α (ng/L): 138.55±10.30 vs. 72.33±10.59, IL-6 (ng/L): 31.35±3.98 vs. 15.75±3.76, both P < 0.01]. Compared with the inflammation model group, the expressions of PD-1/PD-L1 signaling pathway proteins in THP-1 cells in the TFL group were significantly reduced. The expressions of pathway proteins in cells in the TFL+miR-146a inhibitor group were significantly higher than those in the TFL group [PD-1 protein (PD-1/β-actin): 0.85±0.09 vs. 0.37±0.04, PD-L1 protein (PD-L1/β-actin): 0.83±0.08 vs. 0.55±0.06, PI3K protein (PI3K/β-actin): 0.85±0.09 vs. 0.63±0.06, p-Akt protein (p-Akt/Akt): 0.98±0.10 vs. 0.75±0.07, all P < 0.05]. Conclusion:TFL regulates the expression of miR-146a to inhibit the PD-1/PD-L1 signaling pathway in THP-1 cells, regulates the immune barrier of sepsis induced in cell inflammation model in vitro, and thus protects LPS induced THP-1 cells.

Autophagy is a lysosome-dependent intracellular degradation process，and it is a key mechanism of diabetic cardiomyopathy （DCM）. Autophagy has dual regulatory effects on DCM. Under physiological conditions，normal autophagy can promote the decomposition of damaged cardiomyocytes and metabolites，so as to reduce the damage of harmful substances to the body and provide energy for cardiomyocytes. Under pathological conditions，the inhibited autophagy of cardiomyocytes will cause the accumulation of damaged cells and metabolites，which will cause damage to cardiomyocytes and eventually aggravate cardiac dysfunction in the patients with DCM. However，the over autophagy of cardiomyocytes will lead to autophagic death of a large number of cardiomyocytes and result in pathological myocardial remodeling and cardiac dysfunction，thus promoting the progression of DCM. Therefore，the restoration of a normal autophagy level is the key means to protect cardiomyocytes and improve the prognosis of DCM. Chinese medicine can regulate autophagy to treat DCM. Specifically，it can promote autophagy （making up for deficiency） or inhibit autophagy （removing excess） to restore the balance of autophagy，thereby alleviating DCM.

Bipolar disorder （BD） is considered to be mainly related to qi， phlegm， fire and deficiency. Binding constraint of liver qi is the initial cause， while phlegm and qi interact obstruction as well as phlegm and fire interact binding is the key pathogenesis of the transformation between depression and mania， and deficiency of both qi and yin is the main reason of the protracted course of disease. In clinical practice， BD is divided into binding constraint of liver qi pattern， phlegm and qi interact obstruction pattern， phlegm and fire interact binding pattern， and deficiency of both qi and yin pattern， which can be treated with Jinyu Shugan Powder （金玉疏肝散）， Kaiyu Wendan Decoction （开郁温胆汤）， Qingxin Huatan Decoction （清心化痰汤）， and Baihe Shengmai Beverage （百合生脉饮） in their modifications respectively； moreover， Guanye Jinsitao （Herba Hyperici Perforati） is usually used to rectify qi， relieve phlegm and clear heat. It is also suggested to put focus on the prevention and treatment of qi， phlegm and heat simultaneously， and modify the medicinals flexibly in accordance with the pathogenesis evolution and the abnormal exuberance.