[Discussion on Acupuncture as adjunctive therapy for chronic stable angina: a randomized clinical trial published in JAMA Internal Medicine].

Professor LIANG Fan-rong's team from Chengdu University of TCM published Acupuncture as adjunctive therapy for chronic stable angina: a randomized clinical trial in JAMA Internal Medicine on July 29, 2019, which demonstrates that acupuncture as an adjunctive therapy is safety and effective for mild and moderate chronic stable angina. Sixteen experts at home and abroad conducted serious discussions on the research design, evaluation methods, principles and mechanisms, clinical significance and enlightenment of future acupuncture research, and provided practical suggestions for acupuncture and moxibustion to go global and gain international recognition.

[Evidence-based medicine research on low-back pain with sham-acupuncture controlled design and the analysis on dynamic acupoint positioning].

To explore the positioning of acupoints, a research was done with PubMed for system reviews and clinical trials on treatment of low-back pain with sham-acupuncture controlled design from January 1, 2010 to October 27, 2017. Six system reviews and 12 sham-controlled acupuncture random trials were found. The statistical difference was not found in all the 6 trials with standard acupoint compared with the sham-acupuncture among the 8 penetrating skin sham-control trials. The statistical difference was found in the two trials with penetrating skin sham control, who was used individualized treatment, twirling for qi arrival or palpation for ashipoint. It is considered that sham-acupuncture penetrating skin is not a placebo, and needling with standard or dynamic acupoint may reduce low-back pain, and dynamic acupoint positioning may be better than standard acupoint positioning.

Facilitating effects of berberine on rat pancreatic islets through modulating hepatic nuclear factor 4 alpha expression and glucokinase activity.

AIM: To observe the effect of berberine on insulin secretion in rat pancreatic islets and to explore its possible molecular mechanism. METHODS: Primary rat islets were isolated from male Sprague-Dawley rats by collagenase digestion and treated with different concentrations (1, 3, 10 and 30 micromol/L) of berberine or 1 micromol/L Glibenclamide (GB) for 24 h. Glucose-stimulated insulin secretion (GSIS) assay was conducted and insulin was determined by radioimmunoassay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to evaluate cytotoxicity. The mRNA level of hepatic nuclear factor 4 alpha (HNF4alpha) was determined by reverse transcription polymerase chain reaction (RT-PCR). Indirect immunofluorescence staining and Western blot analysis were employed to detect protein expression of HNF4alpha in the islets. Glucokinase (GK) activity was measured by spectrophotometric method. RESULTS: Berberine enhanced GSIS rather than basal insulin secretion dose-dependently in rat islets and showed no significant cytotoxicity on islet cells at the concentration of 10 mumol/L. Both mRNA and protein expressions of HNF4alpha were up-regulated by berberine in a dose-dependent manner, and GK activity was also increased accordingly. However, GB demonstrated no regulatory effects on HNF4alpha expression or GK activity. CONCLUSION: Berberine can enhance GSIS in rat islets, and probably exerts the insulinotropic effect via a pathway involving HNF4alpha and GK, which is distinct from sulphonylureas (SUs).

[Effects of berberine on expression of hepatocyte nuclear factor 4alpha and glucokinase activity in mouse primary hepatocytes].

OBJECTIVE: To observe the expression of hepatocyte nuclear factor 4alpha (HNF4alpha) and the activity of key enzyme glucokinase (GK) in glucose metabolism, and further to investigate the possible mechanism of berberine in treating type 2 diabetes. METHOD: Mouse primary hepatocytes were isolated by an improved single two-step perfusion method. The murine hepatocytes were cultured and incubated with berberine (0, 1, 3, 10, 30, 100 micromol x L(-1)) and 1 mmol x L(-1) metformin for 24 h respectively. The mRNA expression of HNF4alpha were quantified by RT-PCR and the protein expression of HNF4alpha were quantified by Western-blot. And the activity of GK were detected with enzyme kinetics method. RESULT: As compared with the negative control group, at a certain concentration range, the expression of HNF4alpha mRNA and protein and the activity of GK were promoted by berberine. Both of them reached the top at the concentration of 30 micromol x L(-1) (P<0.01). But the metformin made no difference with the negative control group on the expression of HNF4alpha and the activity of GK. CONCLUSION: It is suggested that the effects of berberine on improving glucose metabolism can be mechanically associated with its up-regulating the HNF4a expression and inducing the activity of hepatic glucokinase.

[Protective effects of Huanglian Jiedu decoction on vascular endothelial function in type 2 diabetic rats].

OBJECTIVE: To investigate the effects of Huanglian Jiedu (HLJD) decoction on vascular endothelial function in type 2 diabetic rats and explore the prophylactic and therapeutic significance and pharmacological mechanism of HLJD decoction in type 2 diabetic angiopathic complication. METHOD: The murine type 2 diabetes models were induced by the intravenous injection of a small dose of streptozotocin plus high fat and high caloric laboratory chow. Then modeled diabetic animals were divided into model group, HLJD group, and aspirin group. Normal ratsfed with routine chow were designated as normal group. The oral glucose tolerance test (OGTT) were performed in all animals, 9 weeks after treatment, the changes of murine body weights and levels of fasting blood glucose (FBG), serum total cholesterol (TC), triglycerides (TG), high density lipoprotein-cholesterol(HDL-C), fasting serum insulin (FINS), serum nitric oxide, plasma endothelin, angiotensin II and von Willebrand Factor (vWF) were determined 10 weeks after treatment. RESULT: Compared with model group, the result of OGTT of HLJD group was improved. The levels of the body weights, TC, TG and ET in HLJD group weredecreased (P < 0.05). The levels of FBG,INS, AngII and vWFwere significantly decreased (P < 0.01), and the levels of HDL-C and NO were obviously increased (P < 0.05), as compared with those in model group. Furthermore. The levels of FBG was lower in HLJD group than in aspirin group (P < 0.05), and the improvement of TG, HDL-C,NO, AngII, vWF levels in HLJD group was more greatly than that in aspirin group, but there was not significant difference between two groups (P > 0.05). CONCLUSION: It is suggested by the present results that HLJD decoction could protect vascular endothelium from early damage in type 2 diabetes. The protective effects of HLJD on endothelium might be related to its ability of reducing the blood glucose, adjusting plasma lipids profiles, improving insulin resistance, antagonizing inflammatory mediators and inducing endothelium-dependent vascular relaxation.

Induction of pancreatic duct cells of neonatal rats into insulin-producing cells with fetal bovine serum: a natural protocol and its use for patch clamp experiments.

AIM: To induce the pancreatic duct cells into endocrine cells with a new natural protocol for electrophysiological study. METHODS: The pancreatic duct cells of neonatal rats were isolated, cultured and induced into endocrine cells with 15% fetal bovine serum for a period of 20 d. During this period, insulin secretion, MTT value, and morphological change of neonatal and adult pancreatic islet cells were comparatively investigated. Pancreatic beta-cells were identified by morphological and electrophysiological characteristics, while ATP sensitive potassium channels (K(ATP)), voltage-dependent potassium channels (K(V)), and voltage-dependent calcium channels (K(CA)) in beta-cells were identified by patch clamp technique. RESULTS: After incubation with fetal bovine serum, the neonatal duct cells budded out, changed from duct-like cells into islet clusters. In the first 4 d, MTT value and insulin secretion increased slowly (MTT value from 0.024+/-0.003 to 0.028+/-0.003, insulin secretion from 2.6+/-0.6 to 3.1+/-0.8 mIU/L). Then MTT value and insulin secretion increased quickly from d 5 to d 10 (MTT value from 0.028+/-0.003 to 0.052+/-0.008, insulin secretion from 3.1+/-0.8 to 18.3+/-2.6 mIU/L), then reached high plateau (MTT value >0.052+/-0.008, insulin secretion >18.3+/-2.6 mIU/L). In contrast, for the isolated adult pancreatic islet cells, both insulin release and MTT value were stable in the first 4 d (MTT value from 0.029+/-0.01 to 0.031+/-0.011, insulin secretion from 13.9+/-3.1 to 14.3+/-3.3 mIU/L), but afterwards they reduced gradually (MTT value <0.031+/-0.011, insulin secretion <8.2+/-1.5 mIU/L), and the pancreatic islet cells became dispersed, broken or atrophied correspondingly. The differentiated neonatal cells were identified as pancreatic islet cells by dithizone staining method, and pancreatic beta-cells were further identified by both morphological features and electrophysiological characteristics, i.e. the existence of recording currents from K(ATP), K(V), and K(CA). CONCLUSION: Islet cells differentiated from neonatal pancreatic duct cells with the new natural protocol are more advantageous in performing patch clamp study over the isolated adult pancreatic islet cells.

Therapeutic effects of berberine in impaired glucose tolerance rats and its influence on insulin secretion.

AIM: To explore the anti-diabetic effects of berberine and its influence on insulin secretion. METHODS: Impaired glucose tolerance rats induced by iv injection of streptozotocin 30 mg/kg were treated with berberine 187.5 and 562.5 mg/kg while fed with high fat laboratory chow. After rats were treated for 4 weeks, oral glucose tolerance was determined, and for 8 weeks, the fasting blood glucose, insulin, lipid series were determined. In insulin secretion experiments, berberine 93.75, 187.5, and 562.5 mg/kg was administered orally to BALB/c mice at a bolus. The murine serum was collected 2 h after the berberine administration for insulin determination. Insulin released from HIT-T15 cells and pancreatic islets incubated with berberine 1-100 micromol/L for 12 h was determined. RESULTS: The levels of fasting blood glucose (7.4+/-1.5 or 7.3+/-1.3 vs 9.3+/-1.3 mmol/L), triglycerides (0.61+/-0.22 or 0.63+/-0.17 vs 1.8+/-0.7 mmol/L), total cholesterol (1.8+/-0.3 or 1.9+/-0.3 vs 2.2+/-0.2 mmol/L), free fatty acid (456+/-93 or 460+/-72 vs 550+/-113 micromol/L) and apolipoprotein B (0.37+/-0.02 or 0.42+/-0.05 vs 0.46+/-0.04 g/L) were reduced greatly in berberine-treated groups at doses of 187.5 and 562.5 mg/g/d, respectively as compared with those in control group (P<0.05 or P<0.01), whereas high density lipoprotein-cholesterol (1.5+/-0.3 or 1.4+/-0.3 vs 1.1+/-0.1 g/L), apolipoprotein AI (0.80+/-0.08 or 0.87+/-0.08 vs 0.71+/-0.06 g/L) were significantly increased (P<0.05 or P<0.01), and oral glucose tolerance was improved. In vitro experiment showed that berberine 1-10 micromol/L facilitated insulin secretion of HIT-T15 cells and murine pancreatic islets in a dose-dependent manner. Meanwhile murine serum insulin level (27.5+/-2.7 or 29+/-4 or 29+/-4 vs 24.3+/-2.8 pIU/L) was undoubtedly promoted and blood glucose (4.52+/-0.31 or 4.45+/-0.29 or 4.30+/-0.19 vs 4.87+/-0.21 mmol/L) was reduced after berberine administration at doses of 93.75, 187.5, and 562.5 mg/kg, respectively in the BALB/c mice. CONCLUSION: Berberine possesses anti-diabetic effects, which is related to the property of stimulating insulin secretion and modulating lipids.