NMN synbiotics intervention modulates gut microbiota and metabolism in APP/PS1 Alzheimer's disease mouse models.

Alzheimer's disease (AD) is a complex neurodegenerative condition with growing evidence implicating the gut microbiota in its pathogenesis. This study aimed to investigate the effects of NMN synbiotics, a combination of ß-nicotinamide mononucleotide (NMN), Lactobacillus plantarum, and lactulose, on the gut microbiota composition and metabolic profiles in APP/PS1 transgenic mice. Results demonstrated that NMN synbiotics led to a notable restructuring of the gut microbiota, with a decreased Firmicutes/Bacteroidetes ratio in the AD mice, suggesting a potential amelioration of gut dysbiosis. Alpha diversity indices indicated a reduction in microbial diversity following NMN synbiotics supplementation, while beta diversity analyses revealed a shift towards a more balanced microbial community structure. Functional predictions based on the 16S rRNA data highlighted alterations in metabolic pathways, particularly those related to amino acid and energy metabolism, which are crucial for neuronal health. The metabolomic analysis uncovered a significant impact of NMN synbiotics on the gut metabolome, with normalization of metabolic composition in AD mice. Differential metabolite functions were enriched in pathways associated with neurotransmitter synthesis and energy metabolism, pointing to the potential therapeutic effects of NMN synbiotics in modulating the gut-brain axis and synaptic function in AD. Immunohistochemical staining observed a significant reduction of amyloid plaques formed by Aß deposition in the brain of AD mice after NMN synbiotics intervention. The findings underscore the potential of using synbiotics to ameliorate the neurodegenerative processes associated with Alzheimer's disease, opening new avenues for therapeutic interventions.

Impact of Donepezil Supplementation on Alzheimer's Disease-like Pathology and Gut Microbiome in APP/PS1 Mice.

Based on published information, the occurrence and development of Alzheimer's disease (AD) are potentially related to gut microbiota changes. Donepezil hydrochloride (DH), which enhances cholinergic activity by blocking acetylcholinesterase (AChE), is one of the first-line drugs for AD treatment approved by the Food and Drug Administration (FDA) of the USA. However, the potential link between the effects of DH on the pathophysiological processes of AD and the gut microbiota remains unclear. In this study, pathological changes in the brain and colon, the activities of superoxide dismutase (SOD) and AChE, and changes in intestinal flora were observed. The results showed that Aß deposition in the prefrontal cortex and hippocampus of AD mice was significantly decreased, while colonic inflammation was significantly alleviated by DH treatment. Concomitantly, SOD activity was significantly improved, while AChE was significantly reduced after DH administration. In addition, the gut microbiota community composition of AD mice was significantly altered after DH treatment. The relative abundance of Akkermansia in the AD group was 54.8% higher than that in the N group. The relative abundance of Akkermansia was increased by 18.3% and 53.8% in the AD_G group and the N_G group, respectively. Interestingly, Akkermansia showed a potential predictive value and might be a biomarker for AD. Molecular docking revealed the binding mode and major forces between DH and membrane proteins of Akkermansia. The overall results suggest a novel therapeutic mechanism for treating AD and highlight the critical role of gut microbiota in AD pathology.

Nicotinamide mononucleotide improves the Alzheimer's disease by regulating intestinal microbiota.

Alzheimer's disease (AD) is the most common progressive neurodegenerative disease, and the intestinal flora and its metabolites play an important role in the amelioration of central nervous system (CNS) disorders such as AD through a bidirectional interaction between the gut-brain axis (GBA). Nicotinamide mononucleotide (NMN), one of the precursors for nicotinamide adenine dinucleotide (NAD+) synthesis, reduces the brain features of AD, including neuroinflammation, mitochondrial abnormalities, synaptic dysfunction, and cognitive impairment. However, the impact of NMN on the gut flora of AD is still unknown. In the current study, we investigated the relationship between gut flora and NMN treatment in APP/PS1 transgenic (AD) mice through the 16S ribosomal RNA (rRNA) high-throughput sequencing analysis of mouse feces after being treated with NMN for 16 weeks. The results show that the NMN significantly changed the intestinal microbial community composition in AD mice. The NMN also increased the relative abundance of short-chain fatty acids (SCFAs)-producing bacteria such as Lactobacillus and Bacteroides at the genus level by protecting intestinal health and improving AD. The overall results suggest novel therapeutic strategies for treating AD and highlight the critical role of gut microbiota in AD pathology, and layout the further research.

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.

Intestinal Microflora Changes in Patients with Mild Alzheimer's Disease in a Chinese Cohort.

BACKGROUND: Understanding the relationship between Alzheimer's disease (AD) and intestinal flora is still a major scientific topic that continues to advance. OBJECTIVE: To determine characterized changes in the intestinal microbe community of patients with mild AD. METHODS: Comparison of the 16S ribosomal RNA (rRNA) high-throughput sequencing data was obtained from the Illumina MiSeq platform of fecal microorganisms of the patients and healthy controls (HC) which were selected from cohabiting caregivers of AD patients to exclude environmental and dietary factors. RESULTS: We found that the abundance of several bacteria taxa in AD patients was different from that in HC at the genus level, such as Anaerostipes, Mitsuokella, Prevotella, Bosea, Fusobacterium, Anaerotruncus, Clostridium, and Coprobacillus. Interestingly, the abundance of Akkermansia, an emerging probiotic, increased significantly in the AD group compared with that in the HC group. Meanwhile, the quantity of traditional probiotic Bifidobacteria of the AD group also rose. CONCLUSION: These alterations in fecal microbiome of the AD group indicate that patients with mild AD have unique gut microbial characteristics. These specific AD-associated intestinal microbes could serve as novel potential targets for early intervention of AD.

Human Adipose Tissue Cryopreservation: Impact of Different Calf Serum Concentrations on Tissue Viability.

Adipose tissue, which is harvested in large quantities during liposuction, has no blood supply and becomes necrotic within a few hours, if not immediately transplanted. Cryopreservation of adipose tissue allows these samples to be stored and used in diverse fundamental experiments, especially in fat-grafting animal tests that could provide a theoretical basis for clinical applications. Traditionally, fetal bovine serum (FBS) has been added as a cryoprotectant (CPA) to maintain the maximum viability of different tissues after freezing and thawing. Calf serum (CS) comes from the same species as FBS but is more economical compared with FBS-containing medium. The optimal concentration of CS in CPA for banking adipose tissue has not been studied. Here, we studied the cell survival rate, cell viability, tissue structural integrity, number of adipose-derived stem cells and blood vessels, and survival after transplantation into nude mice via ultrastructural evaluation of adipose tissue cryopreserved for 6 months in condition A (60% CS, 15% dimethyl sulfoxide [DMSO], 25% Dulbecco's modified Eagle's medium [DMEM]) and condition B (30% CS, 15% DMSO, 55% DMEM). Our results indicate that CS in addition to CPA results in adequate preservation of adipose tissue, especially when a higher concentration of CS (60%) is used in the CPA.

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.

Analysis of factors related to fertility after endometriosis combined with infertility laparoscopic surgery.

To investigate the influence factors of laparoscopic postoperative pregnancy of patients with endometriosis and infertility, further validate the application of EFI scoring system in endometriosis, and to improve the pregnancy rate.A total of 258 patients with endometriosis and infertility who underwent laparoscopic surgery and follow-up treatment at Wuxi Maternal and Child Health Hospital from January 2015 to December 2016 were selected and divided into pregnant and non-pregnant groups according to whether they were pregnant. All patients were divided into 4 groups according to EFI score: group with EFI score ≥9, 7-8, 4-6, and <4, and divided into I, II, III, and IV groups according to AFS stages. The uterus-laparoscopic surgery was performed. The patients were followed up for 3 years. The factors affecting the pregnancy rate were analyzed. The pregnancy rate and pregnancy types were calculated at different time points.Multivariate analysis showed that age <35 years, infertility time <5 years, secondary infertility, EFI score, postoperative ART application were protection factors of postoperative pregnancy. The 3-year cumulative postoperative pregnancy rate was 75.6%. The cumulative pregnancy rate was 92.2% in group with EFI score ≥9, 85.9% in group with EFI score 7-8, 62.5% in group with EFI score 4-6 and 5.9% in group with EFI score <4, there was significant difference between the 4 groups (P < .05). The proportion of pregnancies in 6 months and 12 months was higher in patients with EFI score ≥7, 61.0% in patients with EFI score ≥9 and 41.1% in patients with EFI score ≥7. The highest natural pregnancy rate was 83.1% in group with EFI score ≥9, and there was significant difference between the 4 groups (P < .05).Age <35 years, infertility time <5 years, secondary infertility, EFI score and ART application were the protective factors of postoperative pregnancy. EFI score had positive significance in predicting and guiding the postoperative pregnancy of patients with endometriosis and infertility. According to EFI score, the pregnancy rate of patients with endometriosis and infertility can be significantly improved by strict management and active pregnancy program.

Transcriptome analysis in normal human liver cells exposed to 2, 3, 3', 4, 4', 5 - Hexachlorobiphenyl (PCB 156).

BACKGROUNDS: Polychlorinated biphenyls are persistent environmental pollutants associated with the onset of non-alcoholic fatty liver disease in humans, but there is limited information on the underlying mechanism. In the present study, we investigated the alterations in gene expression profiles in normal human liver cells L-02 following exposure to 2, 3, 3', 4, 4', 5 - hexachlorobiphenyl (PCB 156), a potent compound that may induce non-alcoholic fatty liver disease. METHODS: The L-02 cells were exposed to PCB 156 for 72 h and the contents of intracellular triacylglyceride and total cholesterol were subsequently measured. Microarray analysis of mRNAs and long non-coding RNAs (lncRNAs) in the cells was also performed after 3.4 µM PCB 156 treatment. RESULTS: Exposure to PCB 156 (3.4 µM, 72 h) resulted in significant increases of triacylglyceride and total cholesterol concentrations in L-02 cells. Microarray analysis identified 222 differentially expressed mRNAs and 628 differentially expressed lncRNAs. Gene Ontology and pathway analyses associated the differentially expressed mRNAs with metabolic and inflammatory processes. Moreover, lncRNA-mRNA co-expression network revealed 36 network pairs comprising 10 differentially expressed mRNAs and 34 dysregulated lncRNAs. The results of bioinformatics analysis further indicated that dysregulated lncRNA NONHSAT174696, lncRNA NONHSAT179219, and lncRNA NONHSAT161887, as the regulators of EDAR, CYP1B1, and ALDH3A1 respectively, played an important role in the PCB 156-induced lipid metabolism disorder. CONCLUSION: Our findings provide an overview of differentially expressed mRNAs and lncRNAs in L-02 cells exposed to PCB 156, and contribute to the field of polychlorinated biphenyl-induced non-alcoholic fatty liver disease.

Ultrasensitive "signal-on" electrochemical aptasensor for assay of acetamiprid residues based on copper-centered metal-organic frameworks.

In present work, a versatile "signal-on" electrochemical aptasensor with ultra-sensitivity and high selectivity for detecting acetamiprid residues has been successfully constructed. Electrochemistry behaviors of as-synthesized copper-centered metal-organic frameworks (CuMOF) on various electrodes were investigated in details. The results indicated that CuMOF exhibited well-behaved redox events. Thus, we used Au-CuMOF as signaling element to label probe DNA (pDNA). The gold nanoparticles-reduced graphene oxide (Au-rGO) has a high specific surface area and excellent conductivity, which was utilized to immobilize complementary strand (cDNA). In the presence of acetamiprid, Au-CuMOF-labeled pDNA would hybridize with the exposed cDNA, allowing CuMOF to approach the electrode and produce a sensitive signaling current. Such a "signal-on" method does not suffer from the drawbacks of "signal-off" methods. The linear range of this proposed electrochemical aptasensor was 0.1 pM-10.0 nM and the detection limit was as low as 2.9 fM. This platform exhibited wonderful selectivity, stability, and repeatability, and was successfully applied to detect acetamiprid residues in tea samples exhibiting enormous practical application potential.

Decoration of a Poly(methyl vinyl ether-co-maleic anhydride)-Shelled Selol Nanocapsule with Folic Acid Increases Its Activity Against Different Cancer Cell Lines In Vitro.

Due to the low therapeutic index of different chemotherapeutic drugs used for cancer treatment, the development of new anticancer drugs remains an intense field of research. A recently developed mixture of selenitetriacylglycerides, selol, was shown to be active against different cancer cells in vitro. As this compound is highly hydrophobic, it was encapsulated, in a previous study, into poly(methyl vinyl ether-co-maleic anhydride)-shelled nanocapsules in order to improve its dispersibility in aqueous media. Following this line of research, the present report aimed at enhancing the In Vitro activity of the selol nanocapsules against cancerous cells by decorating their surface with folic acid. It is known that several cancer cells overexpress folate receptors. Stable folic acid-decorated selol nanocapsules (SNP-FA) were obtained, which showed to be spherical, with a hydro-dynamic diameter of 364 nm, and zeta potential of -24 mV. In comparison to non-decorated selol nanocapsules, SNP-FA presented higher activity against 4T1, MCF-7 and HeLa cells. Moreover, the decoration of the nanocapsules did not alter their toxicity towards fibroblasts, NIH-3T3 cells. These results show that the decoration with folic acid increased the toxicity of selol nanocapsules to cancer cells. These nanocapsules, besides enabling to disperse selol in an aqueous medium, increased the toxicity of this drug In Vitro, and may be useful to treat cancer in vivo, potentially increasing the specificity of selol towards cancer cells.

Characterization of human S-adenosyl-homocysteine hydrolase in vitro and identification of its potential inhibitors.

Human S-adenosyl-homocysteine hydrolase (SAHH, E.C.3.3.1.1) has been considered to be an attractive target for the design of medicines to treat human disease, because of its important role in regulating biological methylation reactions to catalyse the reversible hydrolysis of S-adenosylhomocysteine (SAH) to adenosine (Ado) and l-homocysteine (Hcy). In this study, SAHH protein was successfully cloned and purified with optimized, Pichia pastoris (P. pastoris) expression system. The biological activity results revealed that, among the tested compounds screened by ChemMapper and SciFinder Scholar, 4-(3-hydroxyprop-1-en-1-yl)-2-methoxyphenol (coniferyl alcohol, CAS: 458-35-5, ZINC: 12359045) exhibited the highest inhibition against rSAHH (IC50= 34 nM). Molecular docking studies showed that coniferyl alcohol was well docked into the active cavity of SAHH. And several H-bonds formed between them, which stabilized coniferyl alcohol in the active site of rSAHH with a proper conformation.

Photodynamic Therapy treatment of onychomycosis with Aluminium-Phthalocyanine Chloride nanoemulsions: A proof of concept clinical trial.

The conventional treatment of onychomycosis, a common fungal infection, consists in the use of local and systemic drugs for 4-6 months. This long protocol is often ineffective due to patient compliance, and usually promotes important collateral effects such as liver and kidney failure. As the alternative, Photodynamic Therapy (PDT) has been used as a noninvasive alternative local treatment for onychomycosis due to the reduction of systemic side effects, fact indicates their use for patients undergoing other systemic treatments. In the present article, we evaluated the effectiveness, as well as the safety of PDT mediated by Aluminium-Phthalocyanine Chloride, entrapped in nanoemulsions, as a drug carrier, to treat onychomycosis in a proof of concept clinical trial. To the date, this is the first published clinical trial that uses PDT mediated by nanomedicines to treat onychomycosis. As main results, we can highlight the safety of the clinical protocol and the antifungal effectiveness similar to the conventional treatments. We observed the (1) clinical cure of 60% of treated lesions; (2) the absence of local and systemic adverse effects; (3) from these clinically healed lesions, 40% were negative for fungal infection in laboratorial exams; and (4) nails that presented negative fungal culture were kept without fungal infection for at least four weeks. The innovation of this approach is the absence of collateral effects, due to the local therapeutically treatment, and the possibility to repeat the treatment without inducing fungal resistance, a fact that indicates this approach as a possible alternative protocol for onychomycosis management.

Characterization of S-adenosylhomocysteine/Methylthioadenosine nucleosidase on secretion of AI-2 and biofilm formation of Escherichia coli.

S-adenosylhomocysteine/Methylthioadenosine nucleosidase (SAHN E.C.3.2.2.9) does not exist in mammalian cells but is essential for methyl recycling in numerous bacterial and protozoan species. Inhibition of this enzyme could limit synthesis of autoinducers of bacterial quorum sensing (QS), and hence, causes reduction in biofilm formation and may attenuate virulence. In this study, sahn deletion mutant of E. coli MG1655, sahn-complemented strain, and SANH-overexpressing strain were established and used to identify the secretion of autoinducer-2 (AI-2) and biofilm formation. The results indicated that deletion of the sahn gene abolished the production of the QS signal AI-2 and biofilm formation in mutant strain MG1655-Δsahn. And the complementation strain MG1655-Δsahn (pET-28a-sahn) showed restored production of AI-2 and biofilm formation, which indicates that the sahn gene plays an important role in bacterial quorum sensing. The recombinant SAHN protein was overexpressed and purified. The enzymatic activity of SAHN was successfully determined by a coupling-enzyme analysis based on xanthine oxidase, with the Vmax and Km of SAHN enzymatic reaction confirmed. Given that sahn is essential for the quorum sensing of both Gram-negative and Gram-positive bacteria, SAHN could be a potential target for wide-spectrum antibiotics.

Nanostructured Systems for the Organelle-specific Delivery of Anticancer Drugs.

Nanotechnology has provided powerful tools to improve the chemotherapy of cancer. Different nanostructures have been developed which deliver the anticancer drugs more selectively to tumor than to healthy tissues. The result has generally been the increase in efficacy and safety of classical anticancer drugs. In recent years, several studies have focused not only on the delivery of anticancer drugs to tumors, but also on delivering the drugs to specific organelles of cancer cells. Endoplasmic reticulum, Golgi apparatus, lysosomes, mitochondria, and nucleus have been the targets of different nanostructured drug delivery systems developed with the goal of circumventing drugresistance, increasing drug efficacy, and so on. So far, the results described in the literature show that this strategy may be used to improve chemotherapy outcomes. In this review a discussion is presented on the strategies described in the literature to deliver anticancer drugs to specific organelles of cancer cells by using nanostructures.

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.

Influence of catgut implantation at acupoints on leptin and insulin resistance in simple obesity rats.

OBJECTIVES: To investigate the influence of catgut implantation at acupoints on leptin resistance (LR) and insulin resistance (IR) in the simple obesity rat. METHODS: Rats were made obese with high-fat diets, after which surgical catgut was implanted at Zusanli (ST 36) and Neiting (ST 44) acupoints once a week for 4 weeks (implantation group). Rats from the implantation group were compared with normal rats and unoperated obese rats (control group). Gene expression of the leptin receptor (OB-R) was evaluated using RT-PCR and northern blot. Serum and hypothalamus leptin and insulin (INS) levels were determined by radioimmunoassay. Body weight, Lee's index, body fat, serum and hypothalamus leptin and INS levels, and hypothalamic OB-R gene expression were determined before and after treatment. RESULTS: Body weights, Lee's index, body fat, and serum leptin and INS levels were significantly higher in obese than in normal rats. Hypothalamic leptin and INS levels and OB-R gene expression were significantly lower in obese rats. Catgut implantation at acupoint promoted weight loss and decreased serum leptin and INS levels. Hypothalamic leptin and INS levels and OB-R gene expression increased significantly. CONCLUSIONS: Catgut implantation at acupoint adjusts central and peripheral leptin and promotes hypothalamic OB-R gene expression. This may be an important method for regulation of LR, IR and abnormal endocrinology and metabolism.

Lipopolysaccharide (LPS) detoxification of analogue peptides derived from limulus anti-LPS factor.

Lipopolysaccharide (LPS) plays a critical role in the pathogenesis of sepsis due to gram-negative bacterial infections. Therefore, LPS-neutralizing molecules could have important clinical applications. Our previous work showed, CLP19, an analogue peptide derived from limulus anti-LPS factor (LALF), possessed the capacity to neutralize LPS and thereby inhibit the LPS-induced responses. However, potential cytotoxicity of CLP19 was also found, especially when added to human red blood cells. Accordingly we further developed two peptides (designated as CLP19-1 and CLP19-2) by single- and double-point amino acid substitution of CLP19, respectively, in order to reduce its toxicity and meanwhile retain the anti-LPS activity. In this study, the LPS-detoxifying effectiveness of these peptides was evaluated both in vitro and in vivo. CLP19-1 was found to dose-dependently neutralize LPS in vitro, with significantly lower hemolysis of red blood cells as compared with CLP19. Further in vivo tests verified that CLP19-1 exerted significant protective effects on mice against LPS, characterized by significantly improved survival, decreasing of tumor necrosis factor alpha (TNF-α) serum level and alleviation of tissue injury. Our work indicates that CLP19-1 is worthy of further study as potential anti-LPS agents for the management of sepsis.