New cycloalkyl[b]thiophenylnicotinamide-based α-glucosidase inhibitors as promising anti-diabetic agents: Synthesis, in silico study, in vitro and in vivo evaluations.

In the present study, a series of cycloalkyl[b]thiophenylnicotinamide derivatives against α-glucosidase were synthesized, and evaluated for their in vitro and in vivo anti-diabetic potential. Most of the synthetic analogues exhibited superior α-glucosidase inhibitory effects than the standard drug acarbose (IC50 = 258.5 µM), in which compound 11b with cyclohexyl[b]thiophene core demonstrated the highest activity with an IC50 value of 9.9 µM and showed higher selectivity towards α-glucosidase over α-amylase by 7.4-fold. Fluorescence quenching experiment confirmed the direct binding of 11b with α-glucosidase, kinetics study revealed that 11b was a mixed-type inhibitor, and its binding mode was analyzed using molecular docking. Moreover, analogs compounds 6a-9b, 11b, 12b did not show in vitro cytotoxicity against LO2 and HepG2 cells. Finally, compound 11b exhibited in vivo hypoglycemic activity by reducing the blood glucose levels in sucrose-loaded rats.

SGLT2 inhibitors for prevention of cardiorenal events in people with type 2 diabetes without cardiorenal disease: A meta-analysis of large randomized trials and cohort studies.

To investigate whether sodium glucose cotransporter 2 inhibitors (SGLT2is) can reduce important cardiorenal endpoints in type 2 diabetic adults without established cardiovascular disease (ECD), in those without heart failure (HF), and in those without chronic kidney disease (CKD). We searched PubMed, Embase, Cochrane Central Register of Controlled Trials (CENTRAL) and clinicaltrials.gov. Event-driven randomized controlled trials (RCTs) and cohort studies were included. We conducted random-effects meta-analysis, respectively based on RCTs and cohort studies, on eight cardiorenal endpoints in three type 2 diabetic subgroups. Thirteen large studies were included. Meta-analysis of RCTs showed the high quality evidences: compared with placebo, SGLT2is significantly reduced the risk of major adverse cardiovascular events, cardiovascular death or hospitalization for HF, and progression of CKD in type 2 diabetic adults without ECD [HRs (95 % CIs): 0.88 (0.82, 0.94), 0.76 (0.70, 0.82), and 0.59 (0.52, 0.66), respectively; risk differences (95 % CIs): -1.6 (-2.4, -0.8), -2.6 (-3.3, -2.0), and -2.4 (-2.8, -2.0) per 1000 patient-years, respectively], in those without HF [HRs (95 % CIs): 0.89 (0.82, 0.95), 0.74 (0.67, 0.81), and 0.61 (0.55, 0.67), respectively; risk differences (95 % CIs): -1.7 (-2.9, -0.8), -5.8 (-7.3, -4.2), and -2.3 (-2.6, -1.9) per 1000 patient-years, respectively], and in those without CKD [HRs (95 % CIs): 0.88 (0.82, 0.94), 0.77 (0.71, 0.83), and 0.63 (0.57, 0.70), respectively; risk differences (95 % CIs): -2.4 (-3.6, -1.2), -6.1 (-7.6, -4.5), and -2.2 (-2.6, -1.8) per 1000 patient-years, respectively]. Meta-analysis of cohort studies also showed the benefits of SGLT2is on the three composite outcomes in the three diabetic subgroups. SGLT2is also significantly reduced some other cardiorenal endpoints in these diabetic subgroups. SGLT2is can significantly reduce important cardiorenal events in type 2 diabetic adults without ECD, in those without HF, and in those without CKD; which supports SGLT2is used in these diabetic subpopulations to prevent cardiorenal events.

Clinical efficacy of simple obesity treated by catgut implantation at acupoints.

OBJECTIVE: To observe the clinical effificacy of treatment with catgut implantation at acupoints on simple obesity. METHODS: Following the theory of Chinese medicine (CM), pattern identification (PI) and treatment was based on the patient's symptoms and signs. Patients were observed during three courses and one year following treatment through self-comparison before and after six or seven acupoints catgut implantation. Obesity was divided into fifive types based on PI: (1) Stomach (Wei) and Intestine excess-heat, (2) Spleen (Pi) defificiency and phlegmwet stagnancy, (3) Liver (Gan)-qi stagnation, (4) Spleen-Kidney (Shen) yang deficiency, and (5) Liver-Kidney yin defificiency. Changes in the following measurements were recorded in 820 patients: body weight, body girth, skinfold thickness, body mass index (BMI), fat percentage (F%) and waist/hip ratio (WHR) and in the following blood values: leptin (LP), insulin (INS), blood lipids, fasting blood sugar (FBS), and insulin sensitive index (ISI) before and after the treatment. Values were compared with those of healthy controls (normal group). RESULTS: Catgut implantation showed effificacy with all fifive types of obesity. Effificacy was greater in males than in females. There was no signifificant difference between the different types by Kruskal-Wallis H test, but the effect was best and of the highest number in patients with Stomach and Intestine excess-heat. Skin-fold thickness, body weight, waist circumference, F%, BMI, and WHR in all 820 cases decreased after treatment (at 90 days and one year), with signifificant differences before and after treatment (P<0.01). Improved metabolism of blood lipids was also seen. Following treatment, LP, INS, and FBS decreased signifificantly (P<0.01) and ISI increased signifificantly (P<0.05). CONCLUSION: Catgut implantation at acupoints provided effective and persistent results, convenience, safety, painlessness, and prolonged effect with no side effects, resulting in reduced body weight and fat and improvement in body shape.

[Effects of cuttlefish bone-bone morphogenetic protein composite material on osteogenesis and revascularization of bone defect in rats].

OBJECTIVE: To study the effects of cuttlefish bone-bone morphogenetic protein (BMP) composite material on osteogenesis and revascularization of bone defect in rats. METHODS: The cuttlefish bone was formed into cylinder with the diameter of about 5 mm and height of about 2 mm after the shell was removed, and then it was soaked in the recombinant human BMP 2 to make a cuttlefish bone-BMP (CBB) composite material. Thirty SD rats, with a defect of skull in every rat, were divided into the CBB and pure cuttlefish bone (PCB) groups according to the random number table, with 15 rats in each group. The rats in the group CBB and group PCB were transplanted with the corresponding material to repair the skull defect. At post transplantation week (PTW) 4, 6, and 8, 5 rats from every group were sacrificed by exsanguination, and ink perfusion was performed. One day later, all the transplants and part of the skull surrounding the defect were harvested, and general observation was conducted at the same time. The specimens were paraffin sectioned for HE staining and Masson staining. The area of microvessel and the area of newborn bone were observed and analyzed through histopathological techniques and image collection system. Data were processed with the analysis of variance of factorial design and LSD test. The correlation between the area of microvessel and the area of newborn bone of the group CBB was analyzed with Pearson correlation analysis. RESULTS: (1) The general observation of the transplant region showed that the transplants were encapsulated by a capsule of fibrous connective tissue. The texture of capsule was soft and relatively thick at PTW 4. The texture was tenacious and thin, but rather compact at PTW 6 and 8. The transplants became gelatinous at PTW 4, and similar to the cartilage tissue at PTW 6 and 8. (2) Histological observation showed that the structure of the transplants in two groups was damaged at PTW 4. A moderate quantity of inflammatory cell infiltration could be observed. The amounts of the primary bone trabeculae and microvessels in group CBB were more abundant than those of group PCB, while the number of osteoclasts was less than those of group PCB. At PTW 6, the inflammatory cell infiltration in the transplants in both groups decreased obviously, the cuttlefish bone was found to be further degraded, and the number of newborn microvessels was increased. There were mature bone trabeculae around the transplants in both groups. And there were also mature bone trabeculae in the degraded CBB in group CBB. At PTW 8, the inflammatory reaction in the transplants in both groups disappeared; there were more mature bone trabeculae; the structure of the cuttlefish bone was found to be damaged basically. Bone trabeculae in group PCB were found around the transplant, while the bone trabeculae could be observed not only around the transplant but also in the degraded CBB in group CBB. The amount of the microvessels in group CBB was still larger than that of group PCB. (3) From PTW 4 to 8, the area of microvessel in group CBB [(63 ± 4), ( 136 ± 36), ( 347 ± 31) µm(2)] was larger than that in group PCB [(44 ± 7), (73 ± 4), (268 ± 42) µm(2), P < 0.05 or P < 0.01]. From PTW 4 to 8, the area of newborn bone in group CBB [(236 ± 26), (339 ± 42), (553 ± 40) µm(2)] was larger than that in group PCB [(137 ± 15), (243 ± 21), (445 ± 29) µm(2), with P values all below 0.01]. (4) The relation between the area of microvessel and the area of newborn bone was significantly positive (r = 0.948, P = 0.001). CONCLUSIONS: The CBB may exert good effect on osteogenesis and vascularization of rats with bone defect. It is a good three dimensional scaffold in bone tissue engineering.

A synthetic cyclic peptide derived from Limulus anti-lipopolysaccharide factor neutralizes endotoxin in vitro and in vivo.

Endotoxin, also known as lipopolysaccharide (LPS), is the major mediator of septic shock due to Gram-negative bacterial infections. Recently, much attention has been focused on cationic peptides which possess the potential to detoxify LPS. Limulus anti-LPS factor (LALF), a protein found in the horseshoe crab (Limulus polyphemus), has been proved with striking anti-LPS effects. We synthesized a cyclic peptide (CLP-19), and then investigated its bioactivity both in vitro and in vivo. The ability of CLP-19 to neutralize LPS in vitro was tested using a Limulus amebocyte lysate (LAL) assay and the LPS-binding affinity was measured with an affinity biosensor method. The synthetic peptide LALF31-52 (residues 31 to 52 of LALF) was used as the positive control peptide in this study. It was found that CLP-19 exhibited the significant activity to antagonize LPS without observable cytotoxicity effect on mouse macrophages. CLP-19 directly bound to LPS, and neutralized it in a dose-dependent manner in the LAL assay. Moreover, CLP-19 also showed the remarkable ability to protect mice from lethal LPS attack and to inhibit the LPS-induced tumor necrosis factor alpha (TNF-alpha) release by decreasing serum LPS in vivo. Our work suggests that this peptide is worthy of further investigation as a potential anti-LPS agent in the treatment of septic shock.

Increased interleukin-6 and granulocyte-macrophage colony stimulating factor levels in the sera of patients with non-segmental vitiligo.

BACKGROUND: Although the cause of vitiligo is unknown, an autoimmune theory has been proposed, and there is now convincing evidence that cytokines have an important role in pathogenesis of autoimmunity. OBJECTIVE: To study the possible role of interleukin-1, beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the pathogenesis of vitiligo. METHODS: The authors measured the serum levels of the above-mentioned cytokines from 50 patients with the vitiligo compared with 20 healthy volunteers, employing the method of radioimmunoassay. RESULTS: The results showed that the serum levels of both IL-6 and GM-CSF of the patients with both focal type and generalized type of vitiligo, and the serum level of IL-1 beta of the generalized type,were significantly, higher than those of normal controls in the patients with segmental vitiligo, the serum levels of all the cytokines tested were not significantly different from those of the normal controls. The GM-CSF levels of both focal type and generalized type, and the IL-6 level of the generalized type in progressive stage were significantly higher than those in stable state. CONCLUSION: It is speculated that IL-6 and GM-CSF may be involved in the autoimmune mechanism of non-segmental vitiligo. However, more evidence is required before a definite conclusion can be drawn.