Curcumin-loaded hydroxypropyl-ß-cyclodextrin inclusion complex with enhanced dissolution and oral bioavailability for epilepsy treatment.

Curcumin, the main bioactive component of turmeric, has a wild range of beneficial effects on central nervous diseases, including anti-Alzheimer's disease, antioxidant stress, and anti-inflammation. Currently, it has been demonstrated the anti-epileptic potential. However, curcumin has poor water solubility, high sensitivity to light and heat, and low absorption, which results in low bioavailability and greatly limits the clinical application of curcumin, as well as the elusive effects in anti-epileptic treatment.This study aimed to develop a curcumin hydroxypropyl-ß-cyclodextrin inclusion complex (CUR-HP-ß-CD) to improve its bioavailability and facilitate its potential development as an anti-epileptic drug. The CUR-HP-ß-CD was generated by the solvent evaporation method, which has efficient entrapment, high solubility, and facilitated bioavailability and brain distribution.The solubility of the CUR-HP-ß-CD was 63.5, 60.1, and 52.9 times that of the unformulated curcumin in H2O, HCl (pH 1.2), and PBS (pH 6.8), respectively. The bioavailability of CUR-HP-ß-CD is improved 2.8 times and 38.7 folds higher brain concentrations. Moreover, the therapeutic anti-epileptic effects of CUR-HP-ß-CD were much more effective in pentylenetetrazol (PTZ)-induced zebrafish and mouse models.This study showed a simple and reproducible strategy to effectively improve the bioavailability and therapeutic effects of curcumin, which could be potentially used in epilepsy treatment.

Computational Analysis of the Related Factors of Deep Vein Thrombosis (DVT) Formation in Patients Undergoing Hip Fracture Surgery.

A retrospective study was conducted on 51 patients undergoing hip fracture surgery to investigate the factors associated with the formation of deep venous thrombosis (DVT). The independent sample t-test and correlation analysis were used to sort out and analyze the data. The findings are as follows. (1) Different gender samples showed significant differences in the Caprini score and thrombus location. Most DVTs in females are located in the posterior tibial vein and intermuscular veins. The Caprini score of females was significantly higher than that of males. (2) Age displays a positive correlation with DVT, coronary heart disease, hypertension, and different surgical types, respectively. (3) There is a correlation between age and operation duration. (4) Hyperlipidemia and cerebrovascular disease show a positive correlation with DVT. (5) There was a significant negative correlation between the Caprini score and the quantification of D-dimer. This indicates that in this sample, the higher the patients' Caprini score is, the lower the quantitation of D-dimer will be. (6) Hyperlipidemia and cardiac insufficiency show a positive correlation with cerebrovascular disease. Patients with hyperlipidemia and cardiac insufficiency may also suffer from cerebrovascular diseases.

Effect of auricular electroacupuncture combined with body acupuncture in improving the consciousness of patients after traumatic brain injury: Study protocol for a randomized controlled trial.

BACKGROUND: Traumatic brain injury (TBI) has become a major cause of morbidity and mortality both in China and abroad. Disorders of consciousness following severe TBI is a common refractory complication, resulting in difficult rehabilitation and poor life quality. However, effective therapeutic approaches remain limited. Although acupuncture has been widely applied in the treatment of neurological disorders in China, its efficacy and safety for consciousness recovery remain to be elucidated. METHODS: Here, we conduct a study design and protocol of a randomized, blinded, controlled study to evaluate the efficacy and safety of electroacupuncture at auricular acupoints "heart" and "brainstem" combined with body acupuncture in the consciousness recovery of patients with TBI. A total of 80 patients with initial Glasgow coma scale score between 3 and 8 points will be recruited in the trial and randomized into intervention (combined application of auricular electroacupuncture and body acupuncture) group or control (conventional treatment) group. Patients in the intervention group will receive electroacupuncture at bilateral auricular acupoints "heart" and "brainstem" (4 points in total) combined with body acupuncture in addition to conventional treatment while patients in the control group will receive conventional treatment alone for 8 weeks. The primary outcomes are changes of Glasgow coma scale score and mismatch negativity of event-related brain potentials at baseline after 4 weeks after the final treatment and 4 weeks after the final treatment. The secondary outcome measures will be changes of Barthel and FuglMeyer scores at baseline after 4 weeks after the final treatment and 4 weeks after the final treatment. The safety will also be assessed by monitoring the incidence of adverse events and changes in vital signs during the study. DISCUSSION: Results from this trial will significantly support the application of auricular acupuncture and body acupuncture in the consciousness recovery of patients with severe TBI. If found to be effective and safe, auricular acupuncture combined with body acupuncture will be a valuable complementary option for comatose patients with TBI. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR1800020245. Registered on 21 December 2018.

Therapeutic effect of Fu's subcutaneous needling for hemiplegic shoulder pain among stroke survivors: Study protocol for a randomized controlled trial.

BACKGROUND: Hemiplegic shoulder pain (HSP) is a frequent complication after stroke and limits patients' physical functioning of the affected arm, thus compromising their quality of life. Fu's subcutaneous needling (FSN) has been widely applied in the treatment of pain diseases in China; however, its efficacy and safety for HSP remain to be elucidated. We therefore conducted a randomized, controlled trial to summarize the current evidence on the effects of FSN on the recovery outcomes of stroke survivors with HSP. METHODS: Here, we conduct a study design and protocol of a randomized, blinded, controlled study to evaluate the efficacy and safety of FSN in patients with HSP. A total of 60 patients with numerical rating scale (NRS) score above 1 will be recruited in the trial and randomized into FSN group or usual care (UC) group. Patients in the FSN group will receive FSN treatment combined with UC treatment while patients in the UC group will receive UC treatment alone for 4 weeks. The primary outcomes are changes of NRS at baseline, after the 1st treatment, after the final treatment and 4 weeks after the final treatment. Secondary measurements will be changes of Fugl-Meyer score, constant score, MPQ-SF score, quality of life score, and range of motion at baseline, after the final treatment, and 4 weeks after the final treatment. The safety will also be assessed by monitoring the incidence of adverse events and changes in vital signs during the study. DISCUSSION: Results from this trial will significantly support the application of FSN in the recovery of patients with HSP. If found to be effective and safe, FSN will be a valuable complementary option for patients with HSP. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR1900021644 (registered on March 2, 2019).

Acupuncture for uterine fibroids: Protocol for a systematic review of randomized controlled trials.

BACKGROUND: Uterine fibroids represent the most common gynecological benign tumors in reproductive females. Acupuncture has been applied as a therapeutic modality in China to treat uterine fibroids. However, currently, few critical systematic reviews regarding the effect of acupuncture on uterine fibroids have been published. Our proposed review aims to evaluate the current evidence on the efficacy of acupuncture for uterine fibroids. METHODS: A total of 7 databases were searched from their inception to December 2018, including PubMed, Medline, Embase, the Cochrane Central Register of Controlled Trials, the Chinese National Knowledge Infrastructure database, the Chinese Biomedical database, and the Wanfang database. The primary outcomes will be reduction in uterine volume and number of fibroids. Secondary outcomes are pelvic or low-back pain, assessed by Visual Analog Scale (VAS); Irregular menstrual periods; Low-abdominal pressure symptoms such as frequent or urgent urination, or constipation and adverse events. Data synthesis will be computed by RevManV.5.3.5 software when a data-analysis is allowed. Methodological quality will be evaluated with the risk of bias according to Cochrane Handbook. RESULTS: This study will provide high-quality evidence of acupuncture for uterine fibroids. CONCLUSION: The conclusion of this systematic review will provide evidence to judge whether acupuncture is an effective therapeutic intervention for patients with uterine fibroids. TRIAL REGISTRATION NUMBER: PROSPERO CRD42019120484.

Protective effect and mechanism of lycopene on endothelial progenitor cells (EPCs) from type 2 diabetes mellitus rats.

Endothelial progenitor cells (EPCs), widely existing in bone marrow and peripheral blood, are involved in the repair of injured vascular endothelium and angiogenesis which are important to diabetic mellitus (DM) patients with vascular complications. The number and the function of EPCs are related to the advanced glycation end products (AGEs) generated in DM patients. Lycopene (Lyc) is an identified natural antioxidant that protects EPCs under the microenvironment of AGEs from damage. However, the underlying mechanism remains unclear. To investigate the effect of Lyc on EPCs, we isolated EPCs from DM rat bone marrow and determined cell proliferation, cell cycle,apoptosis and autophagy of EPCs. The present study showed that 10µg/mL Lyc improved cell proliferation and had low cytotoxicity in the presence of AGEs. In addition, Lyc rescued S phase of the cell cycle arrest, reduced apoptosis rate and decreased autophagic reaction including ROS and mitochondrial membrane potential (MMP) of EPCs. Moreover, Lyc combined use of autophagy inhibitors, 3-MA, had better protective effects. Taken together, our data suggests that Lyc promotes EPCs survival and protect EPCs from apoptosis and oxidative autophagy induced by AGEs, further remaining the number and function of EPCs. This study provides new insights into Lyc protective mechanism of AGEs-induced oxidative autophagy in EPCs from DM patients and offers a new therapy for DM vascular complications.

The improvement effects of edible bird's nest on proliferation and activation of B lymphocyte and its antagonistic effects on immunosuppression induced by cyclophosphamide.

Edible bird's nest (EBN) is regarded as an immune-enhancing food in the People's Republic of China. The aim of this study is to demonstrate the efficiency of EBN in improving the immunity of mouse both in vivo and in vitro. We observed the effects of EBN on spleen lymphocytes proliferation and activation, as well as immunoglobulin isotypes as indicators. In addition, we evaluated the content of total sIgA in the intestinal juice to assess mucosal immunity. The results showed that EBN could promote the proliferation and activation of B-cells and increase IgE, IgA, IgM, and IgG3 levels. We also found that EBN extract can promote the secretion of sIgA in the small intestine. Using cyclophosphamide (CY), we established an immunosuppressed mouse model in which we identified a reversal influence on the ratio of CD3(+)/CD19(+) cells, which indicates that EBN also protects B-cells from the damage induced by CY. We also applied polymyxin B to exclude the interference of lipopolysaccharide throughout the experiment. In conclusion, we found that EBN can reduce the intestinal immune injury induced by CY by accelerating the proliferation and activation of B-cells and enhancing antibody secretion of B-cells.

[Chemical constituents from Mylabris phalerata and their cytotoxic activity in vitro].

Ten compounds were isolated from Mylabris phalerata by using preparative HPLC and column chromatography over MCI gel. On the basis of physical-chemical properties, NMR and MS data analysis, the compounds were identified as 5'-[(1 R,2 R,3 S,6R)-1-hydroxymethyl-2-methyl-3,6-epoxycyclohexane-1,2-dicarboximide]- ethyl-2'-methyl-2'-butenoate (1),cantharidin (2), cyclo-(L-Pro-L-Ala) (3), cyclo-(R-Pro-R-Leu) (4), cyclo-(S-Pro-R-Leu) (5), cyclo-(D-Pro-L-Tyr) (6), indole-3-aldehyde (7), 3-indoleacetic acid (8), valerolactam (9), and 4-hydroxyphthalid (10).Compound 1 was a new compound, and compounds 2-10 were obtained from this genus for the first time. Compounds 1-9 were subjected to cytotoxic activity on HCT-116, HepG2, BGC-823, NCI-H1650, A2780 cell lines, and only compound 2 showed inhibitory effect on all cancer cell lines.

Danggui-Shaoyao-San Improves Learning and Memory in Female SAMP8 via Modulation of Estradiol.

Previous studies showed that Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription, could alleviate cognitive dysfunction of Alzheimer's disease (AD) patients. However, the mechanisms remain unclear; we have now examined the effect of DSS on SAMP8 and elucidated the possible mechanism. Animals were treated with DSS for 2 months, and step-down test and Morris water maze (MWM) test were used to evaluated cognitive abilities. The estradiol (E2), NO, and glycine in blood plasma or in hippocampus were detected to explore the possible mechanisms. The latency of SAMP8 in step-down test was shorter than that of age-matched SAMR1, and DSS increased the latency especially in female animals. In MWM test, we got similar results; SAMP8 spent more time to find the platform, and DSS decreased the time before finding the platform, with little effect on swim velocity, during the training sessions. During test session, DSS increased the time spent in target quadrant especially in female SAMP8. In female SAMP8, plasma E2, NO, and glycine were elevated in plasma or hippocampus tissue. In conclusion, DSS could ameliorate deterioration of cognition in SAMP8, especially in female animals. Increasing E2, NO, and glycine might contribute to the cognitive improvement effect of DSS in female SAMP8.

[Advance in studies on effect of paeoniflorin on nervous system].

Paeoniflorin (PF) is the chief active component of paeonia, with diverse pharmacological actions and wide application. Recently, the effect of PF on nervous system has attracted increasingly more attention. According to current study findings, PF can ameliorate the decline of memory and learning capacities in many dementia model animals, and have effect in protecting the cerebral ischemia injury, treating Parkinson's disease, reliving pain and improving neural synapse plasticity. Thought its mechanism has not been clarified, current findings show that adenosine A1 receptor plays an important role, while M cholinergic receptor, opiate receptor, calcium ion channel and NF-KB may also play a part in paeoniflorin's effect on nervous system.

JD-30, an active fraction extracted from Danggui-Shaoyao-San, decreases ß-amyloid content and deposition, improves LTP reduction and prevents spatial cognition impairment in SAMP8 mice.

JD-30 is an active fraction extracted from Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription. We previously showed that JD-30 could alleviate cognitive dysfunction of the mice induced by intracerebroventricular injection of ß-amyloid (Aß). However, data remain scarce on the effect of JD-30 on an Alzheimer's disease (AD) model and the underlying mechanisms are unknown. Further detailed studies on the effects of JD-30 on spatial cognition of senescence-accelerated mouse prone 8 (SAMP8), a suitable rodent model for cognitive impairment of aged subjects were investigated to elucidate the possible mechanisms. Long-term treatment with JD-30 significantly decreased the prolonged latency of SAMP8 in the Morris water-maze test. It also ameliorated the reduction of long-term potentiation (LTP) and reduced the damage of neurons in the hippocampus of SAMP8. Finally, JD-30 decreased the content and deposition of Aß in the brain of SAMP8. The results show that JD-30 improves deterioration of spatial learning and memory in the SAMP8 mouse model, and by decreasing the content and deposition of Aß, neuronal activity and synaptic plasticity improve, suggesting one of the mechanisms involved.

Danggui-Shaoyao-San and its active fraction JD-30 improve Abeta-induced spatial recognition deficits in mice.

AIMS OF THE STUDY: Previous studies showed that Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription, could alleviate cognitive dysfunction of Alzheimer's disease (AD) patients. However, the mechanism and substance basis remain unknown. JD-30 is a fraction extracted from DSS, whose activity we previously was evaluated. beta-Amyloid (Abeta) is believed to be a critical etiological factor of AD. We have now examined the effect of DSS and JD-30 on AD model mice induced by Abeta, and elucidated the possible mechanism. MATERIALS AND METHODS: Mice were intracerebroventricular injected with the aggregated Abeta(25-35) to mimic AD. Groups of mice were treated with DSS or JD-30 by intragastric infusion over 2 weeks, and their spatial learning and memory capacities were measured by the Morris water maze procedure. The mechanisms were investigated by extracellular microelectrode recordings, and also electron microscopy. RESULTS: Our results show that Abeta(25-35) induced impairment of spatial learning and memory in mice, as well as inhibition of long-term potentiation (LTP) in the hippocampus. The impairments were ameliorated by DSS or JD-30 administration. Additionally, JD-30 not only prevented the aggregation of Abeta(25-35), but disrupted aggregated Abeta(25-35) fibrils. CONCLUSION: These results suggest that JD-30 is one of the chief active fractions extracted from DSS by its ability to ameliorate deterioration of cognition, and by blocking and disrupting the aggregation of Abeta so that synaptic plasticity was improved, which may be one of the mechanisms involved.

[Inhibitory effects of pretreatment with Buyanghuanwu decoction on inflammatory cytokine expressions in the kidneys of rats after induction of brain death].

OBJECTIVE: To observe the inhibitiory effects of pretreatment with Buyanghuanwu decoction (BYHWT) on inflammatory cytokine expressions in the kidneys and the level of reactive oxygen species (ROS) by peripheral blood neutrophils of rats after induction of brain death (BD), and to investigate the effect of BYHWT on the improvement of kidney quality from BD donor. METHODS: 30 male Wistar rats were randomly divided into 3 groups: control group, BD model group and BYHWT group. 6 hours after successful onset of brain death,only the BD rats whose mean arterial blood pressure were higher than 80 mmHg were accepted as donors. Kidneys were harvested and peripheral blood was taken from BD rats. RT-PCR was used to detect the expressions of TNF-a and IL-lpfl mRNA. Western blot was adopted to analyze the expressions of both TNF-alpha and IL-lp protein,and the expression of phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK). Reactive oxygen species( ROS) in the peripheral blood neutrophils were labeled with CM-H2DCFDA and then detected with Flow Cytometry. RESULTS: The expressions of both TNF-alpha and IL-1beta mRNA and protein, and the p-p38MAPK proteins all significantly increased in BD group compared with control group (P < 0.01). However, those in BYHWT group statistically decreased compared with BD group (P < 0.05), but they significantly increased in comparison with control group (P < 0.01). There was a close relation between the expression of p-p38MAPK protein and the expressions of both TNF-a and IL-1beta mRNA and protein. ROS level significantly increased in BD group (P < 0.05 ), whereas it significantly decreased in BYHWT group (P < 0.05). There was no statistically significant difference between BYHWT group and control group (P > 0.05). CONCLUSION: Pretreatment of the rats with BYHWT prior to the induction of rat brain death, can significantly suppress the expressions of inflammatory cytokines and ROS level in the kidneys of rats from BD. It might be related to the blockage of key target points in p38MAPK signaling pathway. Therefore pretreatment with BYHWT could hopefully be an ideal way to improve the quality of kidneys from brain dead donors prior to transplantation.

[Effects of Forskolin on proliferation cell-cycle distribution and activation of the murine T lymphocytes in vitro].

OBJECTIVE: To investigate the effects of Forskolin on activation, proliferation, and cell-cycle distribution of murine CD3+ T lymphocytes, and study the mechanisms of its immunosuppressive effect. METHODS: Singel cell suspensions were prepared from murine lymph nodes. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69 activated by Con A, the proliferation index of activated mouse T lymphocytes was analyzed by CFDA-SE staining, the distribution of the cell cycle was analyzed by PI staining. RESULTS: Forskolin (10(-7), 10(-6), 10(-5) M) could inhibit both the expression of CD69 on CD3+ T lymphocytes and T lymphocyte proliferation index stimulated by Con A in a dose-dependent manner. The C0/G1 of T lymphocytes increased but the S, G2/M phase decreased. CONCLUSION: Forskolin can inhibit the activation and proliferation of murine T lymphocytes in vitro, and arrest activated T lymphocytes from G0/G1 to S or G2/M. Forskolin is a potential immunosupressive agent.

[Effect of shuanghuanglian injection on the activation and proliferation of T lymphocytes in mice in vitro].

AIM: To study the effect of shuanghuanglian injection on the activation and proliferation of T lymphocytes in mice in vitro. METHODS: The toxic effect of SHL on lymphocytes in mice was estimated by MTT test; Double-fluorescent plus flow cytometry to analyze the effect of SHL on the activation of T lymphocytes in mice stimulated by ConA in vitro; MTT test and CFDA-SE plus flow cytometry were used to detect the effect of SHL on the proliferation of T lymphocytes in mice induced by ConA in vitro. RESULTS: SHL had little side effect on lymphocytes in mice in vitro; At the mass concentration of 60, 80, 100, 120 mg/L, SHL inhibited the activation of T lymphocytes in mice stimulated by ConA in vitro (P<0.01); Both MTT test and CFDA-SE stain sign that SHL inhibited the activation of T lymphocytes in mice induced by ConA in vitro at the mass concentration of 60, 80, 100, 120 mg/L (P<0.01). CONCLUSION: SHL can inhibit the activation and proliferation of T lymphocytes in mice in vitro.

[Effect of forsythia suspense extract on phagocytosis of peritoneal macrophages and NO production in vitro].

AIM: To investigate the effect of forsythia suspensa (FS) extract on phagocytosis of peritoneal macrophages and NO production in vitro. METHODS: The peritoneal macrophagess were isolated from BALB/c mice. After stained with CFDA-SE, the DH5alpha were co-cultured with peritoneal macrophagess for 3 h. The effect of FS extract on cyto-phagocytesis in vitro was analyzed by flow cytometry. The peritoneal macrophages were stimulated and activated by LPS in vitro. The effect of FS extract on NO production of the peritoneal macrophages in vitro was measured by NO assay kit. RESULTS: FCM analysis showed that FS extract significantly promoted the phagocytosis of peritoneal macrophages at the final concentration of 40, 80, 160 mg/L, respectively (P<0.05). It also decreased the production of NO at different concentration induced by LPS (P<0.05). CONCLUSION: FS extract can promote phagocytosis of peritoneal macrophages and inhibit NO production in vitro.

Effects of red clover extract on the activation and proliferation of mouse T lymphocytes and the NO secretion of mouse macrophages.

The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide (NO) secretion of mouse macrophages stimulated by lipopolysaccharide (LPS) for 24 h was assayed by Griess reagent system. We found that RCE had potent anti-inflammatory effects on mice. RCE had little cell toxic effect on mouse lymphocytes and macrophages. RCE strongly inhibited the excessive production of inflammatory mediators (NO, CD69, CD25, CD71), in a dose-dependent manner, like cyclosporine A injection. RCE could inhibit proliferation of CD3+ T lymphocytes. These data suggested that RCE might exhibit anti-inflammatory effect by inhibiting the activation and proliferation of mouse lymphocytes and the NO secretion of mouse macrophages.

[Paharmacological mechanisms of cellular immunity of overall alkali of tongbiling for the treatment of joint destruction].

AIM: To investigate the effect of the overall alkali of Traditional Chinese Medicine tongbiling(TBL) which comprises brucine and strychnine alkaloids on collagen induced-arthritis(CIA) and study its paharmacological mechanisms of cellular immunity. METHODS: Bovine CII was emulsified in Freund's incomplete adjuvant. Wistar rats were injected with Type II collagen intradermally at the base of the tail. After swelling, CIA groups were, randomly divided into physiological saline group and treatment group. Then the swelling of the rats' hindlimbs was evaluated. The whole body of the rats treated on 35 th days was photographed by mammography X-Ray. 96 joints in erosion scoring system and 100 joints in joint spacing narrow(JSN) scoring system were used to observe the joint destruction of CIA from X-Ray comprehensively and objectively. After the rats were killed, the third proximal claw pad of the right hindlimb and left forelimb were stained by HE dying, Neutrophil, lymphocyte, plasmacyte infiltration and hyperplasia of synoviocytes were assessed. Then MTT and Western blot were used to determine the effect of the overall alkali of TBL on proliferation of Jurkat cells and ERK1/2 phosphorylation of Jurkat cells, respectively. RESULTS: Inflammation of CIA joints was aggravated quickly. The swelling of CIA rats treated by MTX and overall alkali of TBL for 35 days was relieved (P<0.05). MTX and overall alkali of TBL inhibited the hyperplasia of synoviocytes. Overall alkali of TBL inhibited the infiltration of lymphocyteS and plasmacytes. Overall alkali of TBL inhibited the proliferation and ERK1/2 phosphorylation of Jurkat cells. CONCLUSION: Overall alkali of TBL could relieve joint inflammation and destruction of CIA rats by blocking the MAPK cell signalling pathway to inhibit the activation and proliferation of T cells. Our study might provide an experimental basis for treatment of rheumatoid arthritis with overall alkali of TBL.

[Effect of overall alkali of Tongbiling on CD69 expression activated mouse T lymphocytes].

AIM: To investigate the effect of overall alkali of Tongbiling(TBL) on CD69 expression on activated mouse T lymphocytes and its possible mechanism. METHODS: Phorbol 12,13-dibutyrate(PDB) or concanavalin (ConA) were added successively into mouse lymphocytic culture with various concentration of overall alkali TBL. After 24 hours, CD69 expression rate on mouse T lymphocytes activated with PDB or ConA was analyzed by flow cytometry. RESULTS: Overall alkali TBL could significantly down-regulate CD69 expression in a dose-dependent manner. CONCLUSION: Overall alkali TBL can significantly inhibit CD69 expression on activated mouse T lymphocytes. This study provided an experimental basis for application of overall alkali TBL to treatment of rheumatoid arthritis.