Efficacy and safety of high-dose intramuscular vitamin D2 injection in type 2 diabetes mellitus with distal symmetric polyneuropathy combined with vitamin D insufficiency: study protocol for a multicenter, randomized, double-blinded, and placebo-controlled trial.

Background: Distal symmetric polyneuropathy (DSPN) is the most common chronic complication of type 2 diabetes mellitus (T2DM). DSPN may lead to more serious complications, such as diabetic foot ulcer, amputation, and reduced life expectancy. Observational studies have suggested that vitamin D deficiency may be associated with the development of DSPN in T2DM. However, interventional studies have found that low-dose vitamin D supplementation does not significantly improve neuropathy in DSPN. This study aims to evaluate the efficacy and safety of intramuscular injection of high-dose vitamin D (HDVD) in T2DM with DSPN combined with vitamin D insufficiency. Methods and analysis: We will conduct a multicenter, randomized, double-blinded, and placebo-controlled trial in four large hospitals. All eligible participants will be randomly assigned to either the vitamin D2 supplement or placebo control group and injected intramuscularly monthly for 3 months. Additionally, anthropometric measurements and clinical data will be collected at baseline and 3 months. Adverse events will be collected at 1, 2, and 3 months. The primary outcome measure is the change in the mean Michigan Neuropathy Screening Instrument (MNSI) score at baseline and 3 months post-intervention. We will use the gold-standard liquid chromatography-tandem mass spectrometry method to distinguish between 25(OH)D2 and 25(OH)D3 levels. The MNSN score before the intervention will be used as a covariate to compare the changes between both groups before and after the intervention, and the analysis of covariance will be used to analyze the change in the MNSI score after HDVD supplementation. Discussion: Glycemic control alone does not prevent the progression of DSPN in T2DM. Some studies have suggested that vitamin D may improve DSPN; however, the exact dose, method, and duration of vitamin D supplementation are unknown. Additionally, neuropathy repair requires HDVD supplementation to sustain adequate vitamin D levels. This once-a-month intramuscular method avoids daily medication; therefore, compliance is high. This study will be the first randomized controlled trial in China to analyze the efficacy and safety of HDVD supplementation for patients with T2DM and DSPN and will provide new ideas for pharmacological research and clinical treatment of diabetic neuropathy. Clinical trial registration: https://www.chictr.org.cn/, identifier ChiCTR2200062266.

A Clinically Selected Staphylococcus aureus clpP Mutant Survives Daptomycin Treatment by Reducing Binding of the Antibiotic and Adapting a Rod-Shaped Morphology.

Daptomycin is a last-resort antibiotic used for the treatment of infections caused by Gram-positive antibiotic-resistant bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA). Treatment failure is commonly linked to accumulation of point mutations; however, the contribution of single mutations to resistance and the mechanisms underlying resistance remain incompletely understood. Here, we show that a single nucleotide polymorphism (SNP) selected during daptomycin therapy inactivates the highly conserved ClpP protease and is causing reduced susceptibility of MRSA to daptomycin, vancomycin, and ß-lactam antibiotics as well as decreased expression of virulence factors. Super-resolution microscopy demonstrated that inactivation of ClpP reduced binding of daptomycin to the septal site and diminished membrane damage. In both the parental strain and the clpP strain, daptomycin inhibited the inward progression of septum synthesis, eventually leading to lysis and death of the parental strain while surviving clpP cells were able to continue synthesis of the peripheral cell wall in the presence of 10× MIC daptomycin, resulting in a rod-shaped morphology. To our knowledge, this is the first demonstration that synthesis of the outer cell wall continues in the presence of daptomycin. Collectively, our data provide novel insight into the mechanisms behind bacterial killing and resistance to this important antibiotic. Also, the study emphasizes that treatment with last-line antibiotics is selective for mutations that, like the SNP in clpP, favor antibiotic resistance over virulence gene expression.

Five Traditional Chinese Medicine External Treatment Methods Combined with Mecobalamin for Diabetic Peripheral Neuropathy: A Network Meta-Analysis.

Background: Diabetic peripheral neuropathy (DPN) is one of the most common chronic complications of diabetes. Traditional Chinese medicine (TCM) external treatment has been widely used in China as adjunctive treatment, and some small sample clinical studies have proved its effectiveness. However, due to the limited number of studies, we used network meta-analysis to compare the effectiveness of 5 commonly used external treatment methods of traditional Chinese medicine in the treatment of diabetic peripheral neuropathy. Methods: We searched PubMed, EMBASE, The Cochrane Library, Web of Science, CNKI, CBM, WanFang Knowledge Service Platform, and VIP databases and collected and screened randomised controlled trials on the external treatment of traditional Chinese medicine combined with mecobalamin in the treatment of DPN according to the inclusion and exclusion criteria. The search period was from 2011 to May 2021. The quality of included studies was assessed using the revised Cochrane risk-of-bias tool for randomized trials. The outcome indicators are Toronto score, median nerve sensory conduction velocity, and median nerve motor conduction velocity. Results: A total of 22 publications were included in the study. The results of the network meta-analysis showed that acupuncture combined with mecobalamin was superior to other TCM external treatments combined with mecobalamin in terms of decreasing the Toronto score (MD = -2.8, 95% CI: -5.2∼-0.49), improving median nerve sensory conduction velocity (MD = 3.6, 95% CI: 2.4∼4.9), and median nerve motor conduction velocity (MD = 4.5, 95% CI: 2.6∼6.5). The SUCRA value and probability ranking chart showed that among the three outcome indicators, acupuncture combined with mecobalamin was the best, followed by acupoint injection combined with mecobalamin. Conclusion: In this network meta-analysis, acupuncture combined with mecobalamin shows the best results in the treatment of DPN, followed by acupoint injection combined with mecobalamin.

USP9X mRNA expression predicts clinical outcome for esophageal squamous cell carcinoma treated with cisplatin-based therapy.

INTRODUCTION: Combination therapy with cisplatin is the conventional first-line treatment in patients with locally advanced or metastatic esophageal squamous cell carcinoma (ESCC). Ubiquitin-specific protease 9X (USP9X) has been shown to be associated with resistance to chemotherapy drugs in several cancers. The purpose of this study was to explore the predictive effects of USP9X on advanced ESCC patients treated with cisplatin-based regimens. MATERIALS AND METHODS: The subjects were 69 advanced ESCC patients who received first-line cisplatin-based chemotherapy or chemoradiotherapy. The quantitative real-time PCR was performed to measure USP9X mRNA expression. The correlation of USP9X expression with clinical parameters and tumor response was analyzed. The Kaplan-Meier method and Cox analysis were employed to analyze differences in overall survival (OS). RESULTS: USP9X mRNA expression was positively associated with the TMN stage at initial diagnosis. Patients with low USP9X mRNA expression had a significantly higher objective response rate (57.1% vs. 17.6%, P=0.001) and longer median OS (25.0 vs. 14.0 months, P<0.001) than those with high expression in all patients or in different treatment subgroups (all P<0.05). Multivariate analysis showed that low mRNA expression of USP9X emerged as an independent prognostic factor indicating prolonged OS (hazard ratio 0.50, 95% CI 0.34-0.73; P<0.001). CONCLUSION: These findings suggest that high USP9X mRNA expression predicts poor clinical efficacy and survival to cisplatin-based therapy in patients with advanced ESCC.

Green tea polyphenols attenuate hepatic steatosis, and reduce insulin resistance and inflammation in high-fat diet-induced rats.

Non­alcoholic fatty liver disease (NAFLD) is characterized by hepatic steatosis, insulin resistance and inflammation; however, the exact pathogenesis of NAFLD is not fully understood. Green tea polyphenols (GTP) exhibit beneficial effects against metabolic syndrome. However, the effect of GTP on NAFLD remains largely unknown. The aim of the present study was to investigate the effects of GTP on NAFLD in high­fat diet (HFD)­induced rats. The NAFLD rat model was induced with a HFD for 8 weeks. A total of 30 adult male Sprague Dawley rats were randomly divided into three groups: i) Normal control group; ii) HFD group; and iii) HFD with GTP group. Hematoxylin and eosin and Oil Red O analyses were performed. The levels of alanine aminotransferase (ALT), aspartate amino-transferase (AST) and inflammatory cytokines in the serum, as well as oxidative stress markers and hepatic lipids in the liver were measured. In addition, parameters associated with glucose metabolism were also assessed. Western blotting and RT­qPCR were used to determine the expression levels of 5' adenosine monophosphate­activated protein kinase (AMPK). HFD­induced rats exhibited features associated with NAFLD. GTP intervention significantly reduced serum ALT and AST levels. Fasting serum glucose, insulin resistance and hepatic lipid levels were all decreased in the GTP­treated rats. GTP also significantly decreased the levels of TNF­α, IL­6 and malondialdehyde. In contrast, superoxide dismutase levels were increased in the liver. Furthermore, GTP also significantly increased phosphorylation of AMPK and attenuated histopathological changes indicative of injury in liver tissue. GTP has a protective effect on HFD­induced hepatic steatosis, insulin resistance and inflammation, and the underlying mechanism may involve the AMPK pathway.

Effect of preoperative infusion chemotherapy combined with hyperthermia on sPD-L1 and CEA levels and overall survival of elderly patients undergoing radical resection of lung cancer.

PURPOSE: To explore the effect of preoperative infusion chemotherapy combined with hyperthermia on the expressions of sPD-L1 and CEA in elderly patients undergoing radical surgery for lung cancer, and their prognosis. METHODS: 136 elderly patients undergoing radical resection of lung cancer in our hospital from October 2012 to October 2014 were studied. Patients were randomly divided into two groups, namely the combination group and the individual treatment group, with 68 patients in each group. Patients in the individual treatment group received only preoperative chemotherapy, whereas those in combination group received preoperative infusion chemotherapy and preoperative and postoperative hyperthermia. The treatment efficacy, levels of sPD-L1 tumor marker CEA (carcino-embryonic antigen), and T-lymphocyte subsets (CD4+, CD3+, CD8+, CD29+) were compared between the two groups. Three-year follow-up data were collected to compare the overall survival (OS) of the two groups. RESULTS: The effective rates in the combination group and individual treatment group were 87.5 and 67.5%, respectively (p<0.05). After treatment, lower serum levels of CEA and sPD-L1 were seen in the combination group vs the individual treatment group (p=0.036, p=0.008, respectively). Levels of T-lymphocyte subsets CD4+, CD3+, and CD29+ in both groups increased, and were higher in the combination group vs the individual treatment group (p<0.05). Follow-up data demonstrated that OS in the combination group and the individual treatment group was 61.7 and 48.5%, respectively. Significant difference in OS between the two groups was confirmed by Log-rank test (p=0.043). CONCLUSIONS: Preoperative infusion chemotherapy combined with hyperthermia for elderly patients with lung cancer can improve patient immunity, inhibit tumor growth and lengthen overall survival by improving T-lymphocyte subset levels and reducing the circulating tumor cell content.

WenTong HuoXue Cream Can Inhibit the Reduction of the Pain-Related Molecule PLC-ß3 in the Dorsal Root Ganglion of a Rat Model of Diabetic Peripheral Neuropathy.

WenTong HuoXue Cream (WTHX-Cream) has been shown to effectively alleviate clinical symptoms of diabetic peripheral neuropathy (DPN). This study investigated the gene and protein expression of the pain-related molecule PLC-ß3 in the dorsal root ganglion (DRG) of DPN rats. 88 specific pathogen-free male Wistar rats were randomly divided into placebo (10 rats) and DPN model (78 rats) groups, and the 78 model rats were used to establish the DPN model by intraperitoneal injection of streptozotocin and were then fed a high-fat diet for 8 weeks. These rats were randomly divided into the model group, the high-, medium-, and low-dose WTHX-Cream + metformin groups, the metformin group, the capsaicin cream group, and the capsaicin cream + metformin group. After 4 weeks of continuous drug administration, the blood glucose, body weight, behavioral indexes, and sciatic nerve conduction velocity were measured. The pathological structure of the DRG and the sciatic nerve were observed. PLC-ß3 mRNA and protein levels in the DRG of rats were measured. Compared with the model group, the high-dose WTHX-Cream group showed increased sciatic nerve conduction velocity, improved sciatic nerve morphological changes, and increased expression of PLC-ß3 mRNA and protein in the DRG. This study showed that WTHX-Cream improves hyperalgesia symptoms of DPN by inhibiting the reduction of PLC-ß3 mRNA and protein expression in the diabetic DRG of DPN rats.

Correlation of Gli1 and HER2 expression in gastric cancer: Identification of novel target.

HER2 becomes the standard of care for guiding adjuvant treatment of gastric cancer with trastuzumab in recent years. However, the usage of this target agent is still limited because of the resistance to trastuzumab or the negative expression of HER2 in tumor tissues. The Gli1 and HER2 both play an important role in the pathogenesis of gastric cancer. However, the correlation of them is still unclear. Here we found Gli1 and HER2 are highly expressed in gastric cancer tissues, and they are positively related. Next, we found Gli1 positive patients live a shorter survival time no matter HER2 positive or negative. Furthermore, univariate and multivariate analysis revealed that venous invasion, HER2 expression, Gli1 expression were independent prognostic factors for the survival time in gastric cancer. In addition, suppressing the expression level of Gli1 can decrease the cell viability and migration ability in cells and subcutaneous tumors. Finally, we found that HER2 may regulate Gli1 by Akt-mTOR-p70S6K pathway. Inhibit of HER2 and SMO have synergistic effect on reduction of cell viability. In conclusion, Gli1 is a favorable prognostic indicator in gastric cancer. As a novel target, Gli1 worth further study, especially in Her2-targeted therapy-resistant cancers.

Evolutionary and functional characterization of leucoanthocyanidin reductases from Camellia sinensis.

MAIN CONCLUSION: LARs promoted the biosynthesis of catechin monomers and inhibited their polymerization. The accumulation of catechin monomers and polymers was increased by up-regulating the expression of NtLAR and NtANR s in CsMYB5b transgenic tobacco. Tea is rich in polyphenolic compounds, and catechins are the major polyphenols in tea. The biosynthesis of polyphenols is closely related to the expression of the leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR) genes. In this paper, an evolutionary analysis and functional characterization of three CsLARs were performed. The phylogenetic tree showed that plant LARs could be grouped into three, including gymnosperms, monocotyledons and dicotyledons (clusters I and II). The eighth amino acid residue in a conserved LAR-specific motif is changeable due to a transversion (G â†’ T) and transition (G â†’ C) that occur in the corresponding codon. Therefore, plant LARs can be classified as G-type, A-type and S-type LARs due to this variable amino acid residue. Although (2R, 3S)-trans-flavan-3-ols were the products of recombinant CsLARs proteins expressed in Escherichia coli, both (2R, 3S)-trans and (2R, 3R)-cis-flavan-3-ols were detected in tobacco overexpressing CsLARs. However, a butanol/HCl hydrolysis assay indicated that overexpression of the CsLARs caused a decrease in polymerized catechins. A hybridization experiment with CsLARc + AtPAP1 also showed that no polymers other than epicatechin, catechin and glycoside were detected, although the accumulation of anthocyanins was markedly decreased. CsMYB5b promoted the biosynthesis of both flavan-3-ols and proanthocyanidins (PAs). Therefore, LARs promoted the biosynthesis of catechin monomers and inhibited their polymerization. The accumulation of catechin monomers and polymers was increased by up-regulating the expression of the NtLAR and NtANRs in CsMYB5b transgenic tobacco.

Step-up fecal microbiota transplantation (FMT) strategy.

Gut dysbiosis is a characteristic of inflammatory bowel disease (IBD) and is believed to play a role in the pathogenesis of IBD. Fecal microbiota transplantation (FMT) is an effective strategy to restore intestinal microbial diversity and has been reported to have a potential therapeutic value in IBD. Our recent study reported a holistic integrative therapy called "step-up FMT strategy," which was beneficial in treating steroid-dependent IBD patients. This strategy consists of scheduled FMTs combined with steroids, anti-TNF-α antibody treatment or enteral nutrition. Herein, we will elaborate the strategy thoroughly, introducing the concept, potential indication, methodology, and safety of "step-up FMT strategy" in detail.

The effect of treatment with α-glycosidases from Bacteroides fragilis on the survival of rat erythrocytes in the circulation.

BACKGROUND: It has been demonstrated recently that α1,3-galactosidase from Bacteroides fragilis can efficiently convert human group B red blood cells (RBC) to group O cells. In addition, in vitro data indicated that the enzymatic conversion process did not affect the physiological or metabolic parameters of the RBC. The aim of this study was to investigate the lifespan of enzyme- treated RBC in vivo in the circulation. MATERIALS AND METHODS: This was an experimental, randomised study. The rat was selected as the experimental subject because it expresses α-1,3galactosyl on its RBC. The efficiency of Galα1,3Gal epitope removal from RBC treated with α1,3-galactosidase was tested before the transfusion experiment to track the survival of RBC in the circulation. The animals were divided into three groups and injected via the tail vein with native, mock-treated or enzyme-treated RBC labelled with fluorescein isothiocyanate. The survival rates of the fluorescently labelled RBC were monitored by flow cytometry. RESULTS: Flow cytometry showed that α-galactosidase (0.02 mg/mL for RBC with a haematocrit of 30%) efficiently removed Galα1,3Gal epitopes from rat erythrocytes, although small amounts of remaining Galα1,3Gal epitopes were still detected. The in vivo data demonstrated that the half-life of enzyme-treated RBC was a little shorter than that of native RBC. However, the 24-hour survival fractions of native, mock-treated and enzyme-treated RBC were virtually identical. Most importantly, the enzyme-treated RBC, like the native RBC, were still detectable 35 days after transfusion. DISCUSSION: Our results indicate that α-glycosidase treatment had little effect on the in vivo survival kinetics of RBC. These data add further support to the feasibility of translating enzymatic conversion technology into clinical practice.

Effect of different drying methods on the essential oils of mint (Mentha haplocalyx).

The essential oils of fresh, shade-dried, sun-dried, and oven-dried mint of Mentha haplocalyx Brig., and the shade-dried herbs after one hour of soaking were analyzed by GC-MS to provide a scientific basis to regulate the drying methods. Fifty-nine compounds were isolated and identified, including 35 from fresh herbs, 25 from shade-dried herbs, 23 from sun-dried herbs, 17 from oven-dried herbs and 48 from shade-dried mint after one hour of soaking. Eighteen compounds were common to all five samples, including menthol, menthone, and isomenthone, which were the main components. Several of these significantly decreased in shade-dried mint soaked in water. Thus in cleaning and drying processes soaking mint in water should be avoided as far as possible, in case major components are extracted thus producing an inferior product that will undermine its curative effect.

Adverse effect of metformin therapy on serum vitamin B12 and folate: short-term treatment causes disadvantages?

Diabetes is a global public health challenge that imposes heavy burdens on communities and individuals. Metformin, the first-line medication for diabetes, has the superiority of reducing risk of macrovascular diseases, all-cause mortality and even possibly cancers. Recent observational studies, however, have demonstrated that long-term metformin therapy increases the probability of vitamin B12 and folate deficiency, and might contribute to the progression of diabetic peripheral neuropathy. Despite metformin is widely used and extensively studied, randomized controlled trials performed to explore the effects of metformin on vitamin B12 and folate are limited. Besides, whether short-term treatment causes vitamin deficiency is a pending issue. We postulate that even a few-month treatment with metformin results in the decrease of vitamin B12 and folate. However, supplementation of vitamin B12 rather than the combination of vitamin B12 and folate might be profitable based on the mechanism of metformin on vitamins in patients with type 2 diabetes. This viewpoint differs from those of majority that a combined supplementation of vitamin B12 and folate is inclined to be advised.

Impact of vitamin D on chronic kidney diseases in non-dialysis patients: a meta-analysis of randomized controlled trials.

BACKGROUND AND OBJECTIVES: Recent studies have supported a role for both newer and more established vitamin D compounds in improving proteinuria, although systematic evaluation is lacking. Furthermore, concerns remain regarding the influence of vitamin D on the progression of renal function. We analyzed the efficacy and safety of vitamin D in non-dialysis patients and compared the use of newer versus established vitamin D compounds by performing a meta-analysis of randomized controlled trials. DESIGN: A literature search of PubMed (1975 to September, 2012), EMBASE.com (1966 to September, 2012) and Ovid EBM Reviews (through September, 2012) was conducted. RESULTS: Eighteen studies were eligible for final inclusion; of these, six explored the effects of vitamin D on proteinuria, twelve studied the effects of supplementation on renal function, and fifteen discussed the incidence of hypercalcemia. Compared to the placebo or no interference, both the newer and established vitamin D sterols reduced proteinuria to a similar extent (RR, 2.00; 95% CI, 1.42 to 2.81). No decrease in the glomerular filter rate was observed (SMD, -0.10; 95%CI, -0.24 to 0.03), and the risk for dialysis initiation was 1.48 (95% CI, 0.54 to 4.03) with vitamin D treatment. Additionally, there was an increased risk of hypercalcemia for patients treated with either newer or established vitamin D compounds as compared with the controls (RR, 4.78; 95% CI, 2.20 to 10.37). The head-to-head studies showed no differences in the effects of either newer or established compounds on proteinuria or the risk of hypercalcemia. No serious adverse events were associated with the administration of vitamin D. CONCLUSIONS: Vitamin D therapy appears to decrease proteinuria and have no negative influence on renal function in non-dialysis patients. But the occurrence of hypercalcemia should be evaluated when vitamin D is provided. No superiority for newer versus established vitamin D analogue is found.

Effect of antiallergic herbal agents on chloride channel-3 and immune microenvironment in nasal mucosal epithelia of allergic rhinitis rabbits.

BACKGROUND: Allergic rhinitis (AR) is a Th2 dominant cytokine response. Chloride channel-3 (ClC-3) plays an important role in nasal mucosal edema and inflammatory pathologic changes in AR. Antiallergic herbal agents (AHA) are antiallergic herbal products. In the previous study, we have demonstrated that AHA clearly inhibited allergic medium and relieved allergic reaction of AR. The aim of this study was to evaluate the function of ClC-3 and discuss the possible therapeutic effects of AHA on immune microenvironment in AR. METHODS: AHA were produced and used to treat AR. An animal model of an AR rabbit was established by ovalbumin (OVA). The rhinitis rabbits were randomly divided into three groups: AHA treated group (AHATG), model group (MG) and healthy control group (HCG). The expressions of ClC-3 protein were examined by immunohistochemical method. The mucosal epithelial cells of all the rabbit groups were primarily cultured with tissue culture method in vitro with or without rhIL-4 or rhIL-2. Furthermore, the expressions of ClC-3 mRNA were detected by real-time PCR. The levels of monocyte chemotactic factor-1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) protein in culture supernatants were measured by ELISA. RESULTS: The expressions of ClC-3 mRNA increased more in mucosal epithelial cells of MG than those in AHATG and HCG (P < 0.01). The levels of ClC-3 mRNA, MCP-1 and VCAM-1 protein in culture supernatants of MG were significantly higher than those in the other two groups (P < 0.01). Those were significantly increased in MG untreated 12 hours later than those in other two groups (P < 0.01). The expressions of ClC-3 mRNA, MCP-1 and VCAM-1 protein in culture supernatants of MG and HCG treated with rhIL-4 were significantly higher than those in the AHATG treated with rhIL-4 (P < 0.01). The levels of ClC-3 mRNA, MCP-1 and VCAM-1 protein in culture supernatants of all groups treated with rhIL-2 showed no significant changes (P > 0.05). CONCLUSIONS: AHA can inhibit the secretions of ClC-3, MCP-1 and VCAM-1 in mucosal epithelia and improve inflammatory reaction of AR. ClC-3 plays an important role in the secretion of cytokines and mucosal inflammatory response in AR. RhIL-4 can enhance the secretion of ClC-3, MCP-1 and VCAM-1 in mucosal epithelial cells, especially during the AR process. These enhanced effects of rhIL-4 were significantly suppressed by AHA. The secretions of ClC-3, MCP-1 and VCAM-1 can not be induced obviously by rhIL-2 in mucosal epithelial cells in AR.

[Allicin enhances cytotoxicity of CPT-11 to colon cancer LoVo cell in vitro].

OBJECTIVE: To study the effect of allicin on human colon cancer cell line LoVo and the combined effect of allicin and CPT-11 on this cancer cell line. METHOD: The LoVo cells were cultured in vitro and treated with allicin in different concentrations. MTT assay was used to test dynamically the cell growth inhibiting effect. Apoptosis induction (Annexin-V-FITC/PI) and modulation of DNA cell cycle were measured by flow cytometry. The change of cytotoxicity of CPT-11 after combination of allicin at the concentration of 4.0, 8.0 mg x L(-1) were investigated. RESULT: Allicin had inhibitive effect on growth of LoVo cells in a dose and time dependent manner, with IC50 value of 32.23, 10.74, 6.58 mg x L(-1) at 24 h, 48 h and 72 h, respectively. The apoptosis rate of LoVo cells increased progressively as the cells were treated with increasing concentration of allicin in 24 h, while the apoptosis rate achieved peak value when the cells were treated with allicin at the concentration of 8 mg x L(-1) in 48 h. The result indicated the low concentrations of allicin (< 4 mg x L(-1)) lead to G2/M cell cycle arrest, and higer concentrations ( > 4 mg x L(-1)) exert G1 + G2/M cell cycle arrest in 24 h. Compared with single use of CPT-11, the combined use of CPT-11 and allicin (4.0, 8.0 mg x L(-1), respectively) showed increasing cytotoxicity on the LoVo cells, with IC50 of 24 h decreasing from 47.5 to 7.4 and 7.2 mg x L(-1), respectively. CONCLUSION: Allicin has significant anti-proliferation effect on human colon cancer cell line LoVo by induction of apoptosis and arrestment of cell cycle and can enhance the cytotoxicity of CPT-11 on the colon cancer LoVo cell.

Spontaneous activity associated with primary visual cortex: a resting-state FMRI study.

Brain functions during the resting state have attracted considerable attention in the past several years. However, little has been known about spontaneous activity in the sensory cortices in the task-free state. This study used functional magnetic resonance imaging (fMRI) to investigate the existence of spontaneous activity in the primary visual areas (PVA) of normal-sighted subjects and to explore the physiological implications of such activity. Our results revealed that we were able to detect spontaneous activity, which was nonrandom in that it was distinctly clustered both temporally and spatially in the PVA of each subject. In addition, the neural network associated with the PVA-related spontaneous activity included the visual association areas, the precuneus, the precentral/postcentral gyrus, the middle frontal gyrus, the fusiform gyrus, the inferior/middle temporal gyrus, and the parahippocampal gyrus. After considering the functions of these regions, we speculated that the PVA-related spontaneous activity may be associated with memory-related mental imagery and/or visual memory consolidation processes. These findings confirm the presence of spontaneous activity in the PVA and related brain areas. This confirmation supports the perspective that brain is a system intrinsically operating on its own, and sensory information interacts with rather than determines the operation of the system.

B to O erythrocyte conversion by the recombinant alpha-galactosidase.

BACKGROUND: Human group O red blood cells have great benefit in specialized transfusion areas such as armed conflict and natural calamity. The group B antigen differs structurally from group O antigen only by the addition of one terminal alpha-linked galactose residue. In this study we aimed to remove the terminal galactose from group B red blood cell to get group O red blood cell. METHODS: alpha-galactosidase cDNA was cloned by RT-PCR from Catimor coffee beans grown on Hainan Island of China. The vector for alpha-galactosidase cDNA expression was constructed and transferred into Pichia pastoris cells by electroporation. The transgenic cells were cloned by fermentation and the recombinant alpha-galactosidase was purified by ion exchange chromatography. After studying the biochemical characters of alpha-galactosidase, we have used it in converting human erythrocytes from group B to group O. RESULTS: The purity of recombinant alpha-galactosidase was higher than 96%, which was thought to be suitable for the use of blood conversion. Enzymatically converted human group O red blood cells (ECHORBC) exhibited membrane integrity, metabolic integrity, normal cell deformation and morphology. There were no coagulation between ECHORBC and any group of human blood. The ECHORBC will keep normal structure and function for a period of 21 days at 4 degrees C in monoammoniumphosphate nutrient solution. Experiments with Rhesus monkeys and gibbons showed that transfusion of enzymatically converted erythrocytes was safe. CONCLUSION: ECHORBC can be easily obtained from group B red blood cell by alpha-galactosidase digestion. This study suggests that ECHORBC could be transfused to patients safely and efficiently.