Objective To compare the difference between bone marrow stomal cell (BMSC) and adipose-derived stem cell (ADSC) of liver fibrosis in rats.Methods BMSC and ADSC of Sprague-Dawley (SD) rats were isolated and purified.The stem cell markers were detected with flow cytometry.The coculture system was set up with 0.4 μm Transwell insert semipermeable membrane.ADSC or BMSC were co-cultured with hepatic stellate cells (HSC).Normal hepatocyte cell line of rat (BRL) was co-cultured with HSC as negative control group and HSC cultured alone was blank control group.After cultured for 72 hours,the proliferation of HSC was determined by cell counting kit-8 (CCK-8) method.The expression of α-smooth muscle actin (α-SMA) of HSC was detected by Western blotting.The apoptosis of HSC was examined by flow cytometry.After BMSC,ADSC and BRL cultured alone for 72 hours,expression level vascular endothelial growth factor (VEGF),interleukin-10 (IL-10),nerve growth factor (NGF) and transforming growth factor-β1 (TGF-β1) in the culture medium were detected by enzymelinked immunosorbent assay (ELISA) method.The rats model of liver fibrosis were established.The rats were divided into BMSC treatment group,ADSC treatment group,BRL group and culture medium group,six rats in each group,which were injected with 1.5 mL BMSC,ADSC and BRL cells suspension (5 × 106) through portal vein,respectively,and same volume of culture medium was injected to the rate of culture medium group,once every two weeks for four weeks.The pathological changes of liver tissue sections were observed and liver fibrosis markers were tested.T test was performed for comparison between two samples and analysis of variance was used for comparison among multiple groups.Results BMSC and ADSC were successfully isolated and cultured.The phenotype of BMSC and ADSC was similar.Compared with blank control group and negative control group,both ADSC and BMSC could inhibit the proliferation of HSC and promote apoptosis (proliferation,2.43±0.27,2.39±0.33,1.92±0.38 and 2.18±0.31,FBMSC =25.61,FADSC =38.63,both P＜0.05 ;apoptosis rate,(5.59 ± 0.40)％,(6.82±0.57)％,(8.31± 1.03) ％ and (9.36 ± 0.54) ％,FBMSC =73.69,FADSC =97.41,both P＜ 0.05).The effects of ADSC were more significant than those of BMSC (t=5.76 and 5.18,both P＜0.05).There was difference in the cytokine levels secreted by ADSC and BMSC (NGF,(7.46 ± 0.54) pg/mL vs (3.95 ± 0.71) pg/mL,t =10.92,P＜0.05; TGF-β1,(8.79 ±0.93) pg/mL vs (6.36±0.85) pg/mL,t=7.58,P＜0.05).The cell transplantation experiment indicated that both BMSC and ADSC had significant inhibitory effect on liver fibrosis.The activity index of inflammation and degree of fibrosis in BMSC treatment group and ADSCs treatment group were 9.87±2.07,4.17 ± 0.94 and 10.13 ± 1.81,3.98 ± 0.82,which were significantly lower than those in blank control group (13.78±2.53 and 5.09±1.15)and negative control group (13.34± 1.89 and 4.95± 1.22,FBMSC=51.26 and 32.29,P＜0.05; FADSC =46.73 and 40.94,P＜0.05).The level of hyaluronic acid ((191.5±33.2) μg/L and (178.8±28.2) μg/L),type Ⅲ collagen ((19.9±5.1) μg/L and (21.7± 3.3) μg/L) and hydroxyproline ((312.6±38.8) μg/g and (325.8±28.2) μg/g) of BMSC treatment group and ADSC treatment group were significantly lower than those of negative control group and blank control group (hyaluronic acid,(282.3 ± 18.7) μg/L and (287.5 ± 26.7) μg/L),F =73.51 ; type Ⅲ collagen,(35.3± 3.3) μg/L and (32.5±4.3) μg/L,F=76.19; hydroxyproline,(458.4 ± 38.1) μg/g and (473.9 ± 63.7) μg/g,F=60.37,all P＜0.05).However,there was no difference between BMSC treatment group and ADSC treatment (all P＜0.05).Conclusions ADSC and BMSC had similar stem cell characteristics.There was difference in inhibiting the activation of HSC between ADSC and BMSC.But there was no significant difference in inhibiting liver fibrosis of rats in vivo.

Objective To investigate the effect of anti-epileptic soup on the levds of interleukin-2 (IL-2) and interleukin-6 (IL-6) in pentylenetetrazol-induced rats and its mechanism.Methods Healthy adult male SD rats were injected intraperitoneally with pentylenetetrazol kindling 35.0mg/kg to induce a rat epilepsy model.After successful construction of the epilepsy model,the rats were divided into four the groups:blank group,model group,anti -epileptic soup group and sodium valproate (VPA) group.All of the treatments were performed three times a day continuously for 28 days.The rats in the anti-epileptic soup group were given 6ml of anti-epileptic soup.The rats in the VPA group were given 400mg/kg of sodium valproate by garage.The rats in blank group and model group received 6ml of normal saline.After treatment,the seizures level was recorded by using the Diehl six point grading scale,and the expression of serum IL-2 and IL-6 were detected by the enzyme linked immunoassay (ELISA).Results The seizure level of the model group was (4.23 ± 0.43),while that of the anti-epileptic group was (3.47 ± 0.39),indicated that the anti-epileptic soup can reduce the seizures level.The expressions of IL-2 (4.18 ± 0.41)pg/mL and IL-6 (350.0 ± 82.5) pg/mL of the model group were higher than those of the blank group [IL-2 (3.20 ± 0.36) pg/mL,IL-6 (260.2 ± 69.0) pg/mL,P ＜ 0.01],while the expressions of IL-2 (3.48 ± 0.44) pg/mL and IL-6 (287.5 ±74.6) pg/mL of the anti-epileptic group were lower compared with the control group (P ＜0.01).No significant difference was found between the anti-epileptic group and VPA group IL-2 (3.35 ± 0.56) pg/mL and IL-6 (253.8 ± 59.8) pg/mL(P ＞ 0.05).Conclusion The anti-epileptic soup is effective for treating epilepsy in rats,the possible mechanism may be related with downregulating the expression of IL-2 and IL-6.

Objective:To investigate the efficacy of auricular acupoint bean-embedding therapy in the treatment of residual dizziness after successful reduction of benign paroxysmal positional vertigo (BPPV) and its effect on skin sympathetic response.Methods:A total of 110 patients with residual dizziness after successful reduction of BPPV who were admitted to Wenzhou Hospital of Traditional Chinese Medicine from January 2019 to May 2021 were included in this study. They were randomly divided into control and study groups, with 55 patients in each group. Patients in the control group were treated with drugs, and those in the study group were treated with auricular acupoint bean-embedding therapy. Before and after treatment, dizziness handicap inventory (DHI) score, activities-specific balance confidence (ABC) score, and Hamilton Anxiety Scale (HAMA) score were compared between the two groups. The latency and amplitude of sympathetic skin response (SSR) were measured.Results:After 15 days of treatment, DHI scores of all dimensions in the study group were significantly lower than those in the control group ( t = 16.13-20.62, all P < 0.05). ABC score in the study group was significantly higher than that in the control group [(87.90 ± 6.01) points vs. (80.55 ± 8.73) points, t = 3.10, P < 0.05). HAMA score in the study group was significantly lower than that in the control group [(7.85 ± 1.06) points vs. (13.30 ± 2.49) points, t = 8.43, P < 0.001]. The initial latency value of SSR in the study group was significantly higher than that in the control group [(1.95 ± 0.27) ms vs. (1.67 ± 0.21) ms, and the amplitude of SSR in the study group was significantly lower than that in the control group [(1.49 ± 0.15) mV vs. (1.70 ± 0.22) mV, t = 4.73, 4.04, both P < 0.001]. The incidence of adverse reactions in the study group was significantly lower than that in the control group [0.0% (0/55) vs. 10.9% (6/55), χ2 = 4.40, P < 0.05]. Conclusion:Auricular acupoint bean-embedding therapy can effectively alleviate the symptoms of dizziness after successful reduction of BPPV, improve patient's psychological status and autonomic nerve function, which is worthy of clinical promotion.

Objective:To investigate the clinical efficacy of cognitive and behavioral therapy assisted by five-tone therapy for insomnia in sub-healthy people based on digital polysomnography observation.Methods:The clinical data of 70 sub-healthy people with insomnia who received treatment in Wenzhou Hospital of Traditional Chinese Medicine, Zhejiang University of Traditional Chinese Medicine from June 2019 to June 2021 were retrospectively analyzed. These patients were divided into a control group and an observation group ( n = 35 per group) according to different treatment methods. The control group was treated with cognitive and behavioral therapy, while the observation group was treated with cognitive and behavioral therapy assisted by five-tone therapy. Clinical efficacy, syndrome score, sleep quality, quality of life, and digital polysomnography index levels were compared between the two groups. Results:After 4 weeks of treatment, the total effective rate in the observation group was 97% (29/35), which was significantly higher than 83% (34/35) in the control group ( χ2 = 8.77, P = 0.012). The syndrome score in the observation group was (3.26 ± 2.12) points, which was significantly lower than (6.00 ± 1.68) points in the control group ( t = 5.99, P < 0.001). The Pittsburgh Sleep Quality Index score in the observation group was (3.46 ± 2.36) points, which was significantly lower than (5.60 ± 3.35) points in the control group ( t = 3.09, P = 0.003). The WHOQOL-BREF score in the observation group was (67.00 ± 5.26) points, which was significantly higher than (61.54 ± 4.63) points in the control group ( t = 4.61, P < 0.001). Total sleep time, sleep efficiency, and sleep latency in the observation group were superior to those in the control group ( t = 4.56, 5.57, 3.96, all P < 0.05). Conclusion:Cognitive and behavioral therapy assisted by five-tone therapy can markedly reduce insomnia severity and improve the quality of life in sub-healthy people with insomnia. The evaluation of insomnia status based on digital polysomnography indicators can effectively objectively evaluate the causes of insomnia, which is of great significance for guiding clinical treatment.

Objective:To investigate the clinical value of three-dimensional arterial spin marker imaging(3D-ASL) for evaluating collateral circulation compensatory status in patients with cerebral infarction.Methods:A total of 24 patients with cerebral infarction who were treated at Wenzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang University of Chinese Medicine from September 2017 to January 2019 were examined by 1.5-T high-field MR scanner.The changes of cerebral blood flow(CBF) between the infarcted area and the contralateral normal area were compared when the labeled delay time(PLD)=1.5s and PLD=2.5s, and the patients were divided into the group with strong collateral circulation compensation and the group with weak collateral circulation compensation according to the difference in lesion area measured by 3D-ASL(PLD=1.5s and 2.5s). The changes of national institutes of health stroke scale(NIHSS) score and Bathel index at admission and 15 days after admission were compared.Results:3D-ASL measurement of PLD=1.5s showed that the CBF value in the infarcted area was (33.70±20.83)mL/(100g×min), which was significantly lower than that in the contralateral normal area(PLD=1.5s)[(49.93±13.13)mL/(100g×min)]( t=3.229, P<0.05). When PLD=2.5s, ASL measurement results showed that the CBF value in the infarcted area increased significantly[(58.26±23.50)mL/(100g×min) vs.(33.70±20.83)mL/(100g×min)]( t=3.831, P<0.05), and the CBF value in the contralateral normal area also increased significantly[(68.29±14.03)mL/(100g×min) vs.(49.93±13.13)mL/(100g×min)]( t=4.681, P<0.05). At this time, the CBF value in the infarcted area was still significantly lower than that in the contralateral normal area[(58.26±23.50)mL/(100g×min) vs.(68.29±14.03)mL/(100g×min)]( t=1.795, P<0.05). On the day of admission, the BI index of patients in the weak collateral circulation compensation group(12 cases) was significantly higher than that in the strong collateral circulation compensation group(12 cases)[(90.42±10.50) vs.(67.92±27.57)]( t=2.642, P<0.05), and the NIHSS score was significantly lower than that in the strong collateral circulation compensation group[(1.25±1.01) vs.(3.83±3.62)]( t=2.378, P<0.05). After 15 d of admission, there were no significant differences in BI index and NIHSS score between the two groups(all P>0.05). Conclusion:3D-ASL with different delay time can effectively and intuitively reflect collateral circulation compensation of patients with cerebral infarction, and it has very important reference value for the evaluation of patients' disease and the formulation of clinical treatment plan.

Objective To study the mechanism of Sanhuang Decotion in the treatment of ulcerative colitis(UC)under Candida albicans colonization in mice based on Dectin-1-Syk-CARD9 signaling pathway.Methods Mice model of UC with fungal colonization were established with dextran sodium sulfate free drinking and C.albicans intragastric administration.Mice were divided into normal control group,model group,sulfasalazine group,fluconazole group,and Sanhuang Decotion low-and high-dosage groups,and receive corresponding drug interventions.General state of mice were observed,and the disease activity index(DAI)score of mice were calculated.The load of C.albicans in intestine was detected,the length of the colon was measured,and pathological scoring of the colon tissue was performed.The ultrastructural changes of colon epithelium were observed under transmission electron microscopy.The contents of TNF-α,IL-6 and IL-12 in serum and colon tissues were detected by ELISA.The mRNA and protein expression of Dectin-1,Syk,CARD9,NF-κBp65 and inflammation factors in intestinal epithelial cells and colon tissues were detected by qPCR,Western blot and immunohistochemistry.Results Compared with the normal control group,the model group mice showed reduced activity,decreased food intake,accompanied by loose stools,significantly increased DAI score,increased load of C.albicans in the intestine,shortened colon length,and increased histopathological score,with widening of gap between colon epithelial cells,cytoplasmic dissolution,mitochondrial swelling;TNF-α,IL-6 and IL-12 in serum and colon tissue increased,the expressions of Dectin-1 and CARD9 mRNA and protein in colon epithelial cells increased,p-Syk,p-NF-κBp65,CARD9,TNF-α,IL-1β,IL-6 protein expression in colon tissue increased(P<0.01,P<0.05).Compared with the model group,the Sanhuang Decotion high-dosage group mice showed a significant decrease in DAI score,decreased intestinal C.albicans load,increased colon length,decreased histopathological score,more complete and orderly arrangement of microvilli in colon epithelial cells,mild mitochondrial swelling,TNF-α,IL-6 and IL-12 in serum and colon tissue decreased,and the mRNA and protein expression of Dectin-1 and CARD9 in colon tissue increased,the expression of p-Syk,p-NF-κBp65,CARD9,TNF-α,IL-1β,IL-6 protein in colon tissue decreased(P<0.01,P<0.05).Conclusion Sanhuang Decotion may exert an anti C.albicans colonization UC effect by inhibiting the Dectin-1-Syk-CARD9 signaling pathway and reducing the release of inflammatory factors.