Berberine: A Traditional Natural Product With Novel Biological Activities.

CONTEXT: Having been used for thousands of years to treat gastrointestinal diseases, the natural isoquinoline alkaloid, berberine, has exhibited a wide spectrum of biochemical and pharmacological effects in studies of recent years. OBJECTIVE: The review intended to examine the many novel bioactivities of berberine, including antidiabetic, anticancer, neuroprotective, anti-inflammatory, and anti-atherosclerotic actions. DESIGN: The research team searched the MEDLINE database using PubMed, using different keyword combinations, including berberine AND diabetes, berberine AND cancer, berberine AND (neuron OR brain), berberine AND inflammation, and "berberine AND atherosclerosis to find studies evaluating the various effects exerted berberine. CONCLUSION: Berberine is a promising multipotent agent to combat diabetes, cancer, Alzheimer's disease, and other diseases.

Adipose-derived mesenchymal stem cells ameliorate hyperglycemia through regulating hepatic glucose metabolism in type 2 diabetic rats.

Infusion of mesenchymal stem cells (MSCs) has been identified in the rapid alleviation in hyperglycemia of diabetic individuals, but the mechanism involved has not been adequately explained by these cells' potential role in modulating system insulin sensitivity and islet regeneration. In this study, we demonstrated adipose-derived mesenchymal stem cells (ASCs) produced significantly lower blood glucose via promoting hepatic glycogen synthesis and inhibiting hepatic glucose production within 24 h after infusion in T2DM rats. In vitro, HepG2 cells treated with palmitate (PA) were used as a model of hepatic glucose metabolism disorder to confirm that ASCs stimulates the phosphorylation of hepatic AMP-activated protein kinase (AMPK) to restores hepatic glucose metabolism in type 2 diabetes. In summary, this study indicated that ASCs improve hyperglycemia via regulating hepatic glucose metabolism. Additionally, the effect of ASCs on hepatic glucose metabolism depended on the AMPK signaling pathway. Thus, this is the new research of the molecular mechanisms of MSCs administration to improve glucose metabolism, and it may indicate a new treatment target of MSCs in T2DM.

GC-rich promoter elements maximally confers estrogen-induced transactivation of LRP16 gene through ERalpha/Sp1 interaction in MCF-7 cells.

LRP16 gene has been characterized as an estrogen-responsive gene. One 1/2ERE/GC-rich site was previously identified to be indispensable for -676/-214 (region A) fragment within LRP16 regulatory region to confer E2 action. Here, we report that -213/-24 fragment (region B) has higher E2-responsiveness than that of region A in MCF-7 cells, but not in HeLa cells. Deletion and mutation analyses of region B showed that multiple GC-sites are involved in the E2-stimulated response and one 30-bp fragment (-213 to -184 bp) is essential for conferring maximum E2-responsiveness. Results from the cotransfection assays containing Sp1-siRNA revealed that Sp1 is required for the basal transcription activity and E2-responsiveness of both regions A and B. Northern blot analysis demonstrated that inhibition of Sp1 in MCF-7 cells not only decreased the basal expression of LRP16, but markedly impaired its upregulation by E2. Results from gel mobility shift assays exhibited the direct binding of Sp1 protein to the 28-bp fragment (-211 to -184 bp), which was enhanced by the ERalpha titer. Moreover, the functional interaction of ERalpha and Sp1 proteins in the presence of E2 at the GC-rich sites in region B was confirmed by chromatin immunoprecipitation (ChIP) assays. In general, these results demonstrate that GC-rich sites in the proximal promoter of LRP16 gene are sufficient for E2 activation of LRP16 and the -213/-184 fragment containing only one GC site is essential for the maximal induction in MCF-7 cells. We also provide a model for Sp1-dependent regulation of genes by E2 through GC-rich motifs.

[EGFR gene mutation status among lung cancer patients in China].

OBJECTIVE: To investigate EGFR gene mutations in non-small cell lung cancers (NSCLCs) and their correlation with clinicopathologic features and clinical characteristics in Chinese NSCLC patients. METHODS: To analyse EGFR mutations of exons 19 and 21 in NSCLCs by PCR amplification and sequencing. RESULTS: Somatic mutations in TK domain of EGFR were found in 13 cases (17.3%), the majority of mutations were in-frame exon 19 (9.3%) and 6 cases missense mutation in exon 21 (8.0%). The mutation rate was significantly higher in adenocarcinoma (12/31, 38.7%), than in bronchioloalveolar cancer (1/10, 10. 0%), adeno-squamous carcinoma (0/5), pulmonary blastoma (0/2), large cell carcinoma (0/1) and squamous cell carcinoma (0/26). Moreover, mutations were more frequently observed in females (30.0%) than in males (8.9%), and significantly higher in non-smokers (28.2%) than in smokers (5.6%). CONCLUSION: EGFR gene mutation is significantly higher related to adenocarcinomas, females and never-smokers. The results may suggest that a lager portion of adenocarcinomas in Chinese patients, females and non-smokers could be associated with favorable response to gefinib.

[Epidermal growth factor receptor mutation in non-small cell lung cancer and breast cancer].

Somatic mutation in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR) gene is associated with the sensitivity of non-smal cell lung cancer (NSCLC) to TK inhibitor Gefitinib. Mutational analysis for EGFR exons 19 and 21 was performed in 75 NSCLC and 10 breast cancer patients. All patients had not received treatment of Gefitinib. Somatic mutations in TK domain of EGFR were identified in 13 of the 75(13/75, 17.33%) patients, including 7 cases of in-frame deletion in exon 19 (7/75, 9.33%) and 6 cases of amino acid substitution (2573T>G, L858R) in exon 21 (6/75, 8%) . No other mutations were found in 10 breast cancer patients who stained positive for HER2 immunhistochemistry. Adenocarcinoma has a higher rate of mutations than several other types of NSCLC, the mutations occurring more fre-quently in female patients. EGFR mutation rate in Chinese NSCLC patients was higher than that in Caucasians. Our data indicated that Chinese adenocarcinoma patients could benefit from TK inhibitor Gefitinib.

[Effects of interleukin-10 on expression of inflammatory mediators and anti-inflammatory mediators during acute lung injury in rats].

OBJECTIVE: To investigate the effects of interleukin-10 (IL-10) on expression of inflammatory mediators and anti-inflammatory mediators during acute lung injury (ALI). METHODS: Rat model of ALI was reproduced by intratracheal instillation of lipopolysaccharide (LPS) in a dose of 10 mg/kg. Fifty-four male SD rats were randomly divided into control group, LPS group and LPS+IL-10 group, with 18 rats in each group (6 rats at 2, 6 and 24 hours respectively). Arterial gas analysis, the total protein concentration in bronchoalveolar lavage fluid (BALF), the total cell counts and classification in BALF, the lung coefficient, lung pathology were examined. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to determine the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and IL-1ra mRNA in lung tissue. RESULTS: (1)In LPS group the partial pressure of oxygen in artery (PaO(2)) was progressively decreased, while the lung coefficient, the total protein concentration, the total cell counts in BALF were greatly increased, and lung pathology showed severe polymorphonuclear leukocytes infiltration with bleeding and hyaline membranes formation. In LPS+IL-10 group the values of all above parameters were alleviated. (2)In LPS group TNF-alpha mRNA expression peaked at 2 hours, then decreased sharply; IL-1beta mRNA expression greatly increased at 2 hours, peaked at 6 hours, then decreased; IL-1ra mRNA expression increased and peaked at 6 hours, remaining higher than control group at 24 hours. IL-10 inhibited TNF-alpha mRNA and IL-1beta mRNA expression but showed no effect on IL-1ra mRNA expression. CONCLUSION: (1)ALI is characterized by overwhelming expression of TNF-alpha mRNA, IL-1beta mRNA, while the expression of IL-1ra mRNA is much delayed than TNF-alpha mRNA and IL-1beta mRNA. It suggests that there is an imbalance between inflammatory/anti-inflammatory mediators in the early phase of ALI. (2)IL-10 can inhibit the expression of inflammatory mediators and has no effect on the expression of anti-inflammatory mediators, thus it contributes to the balance between them, ameliorating ALI in rats.

Effects of the Four-Herb Compound ANBP on Wound Healing Promotion in Diabetic Mice.

Wound healing is a troublesome problem in diabetic patients. Besides, there is also an increased risk of postsurgical wound complications for diabetic patient. It has been revealed that traditional Chinese medicine may promote healing and inhibit scar formation, while the changes of morphology and physiology of wounds on such medicine treatment still remain elusive. In this study, we first used the ultralow temperature preparation method to produce mixed superfine powder from Agrimonia pilosa (A), Nelumbo nucifera (N), Boswellia carteri (B), and Pollen typhae (P), named as ANBP. Applying ANBP on 40 streptozotocin (STZ)-induced diabetic C57BL/6 mice (4-6 weeks, 20 ± 2 g), we observed that the wound healing process was accelerated and the wound healing time was shortened (14 days, P < .05). Pathological observation using hematoxylin-eosin staining indicated that inflammatory cells were reduced (P < .05) while the thickness of granulation tissue and length of epithelial tongue were increased (P < .05). The vascular density was increased on 7 and 14 days after ANBP treatment. Masson and Sirius red staining showed that, at the early stage of trauma, the expressions of Col I and Col III, especially Col III, were increased in the ANBP group (P < .05). Studies in vitro demonstrated that tubular formation was significantly increased after ANBP treatment on human vascular endothelial cells in a dose-dependent way. Taken together, our studies revealed that ANBP treatment could accelerate wound healing, promote vascularization, and inhibit inflammation, suggesting the potential clinic application of ANBP for diabetes mellitus and refractory wounds.

The four-herb Chinese medicine ANBP enhances wound healing and inhibits scar formation via bidirectional regulation of transformation growth factor pathway.

The four-herb Chinese medicine ANBP is a pulverized mixture of four herbs including Agrimonia Eupatoria (A), Nelumbo Nucifera Gaertn (N), Boswellia Carteri (B) and Pollen Typhae Angustifoliae (P). The combination of the four herbs was first described in Chinese canonical medicine about 2000 years ago for treatment of various trauma disorders, such as hemostasis, antiinflammatory, analgesia, and wound healing, etc. However, the precise mechanisms of ANBP are still unclear. In our study, using rabbit ear hypertrophic scar models of full-thickness skin defect, we showed that local ANBP treatment not only significantly enhanced wound healing by relieving inflammation, increasing formation of granulation tissue and accelerating re-epithelialization, but also reduced scar formation by decreasing collagen production, protuberant height and volume of scars, and increasing collagen maturity. We demonstrated that these effects of ANBP are associated with transforming growth factor (TGF)-ß1-mediated signalling pathways through Smad-dependent pathways. ANBP treatment significantly increased expression of TGF-ß1 and Smad2/3 mRNA at the early stage of wound healing, and led to markedly decrease expression of TGF-ß1 and Smad2/3 compared with the control group after 14 days post-wounding. Taken together, our results defined a bidirectional regulation role of ANBP for TGF-ß1/Smad pathway in promoting wound healing and alleviating scar formation, which may be an effective therapy for human wounds at the earliest stage.

Two Chinese pedigrees for adenomatous polyposis coli: new mutations at codon 1309 and predisposition to phenotypic variations.

Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease caused by a mutation in the adenomatous polyposis coli (APC) gene. Some studies have attempted to correlate mutations at codon 1309 with classic FAP (≥100 colorectal polyps). We report two Chinese FAP pedigrees with new frameshift mutations at codon 1309, in which affected individuals manifest phenotypic variations. Comprehensive physical examinations were performed for all living individuals and the medical data of deceased patients were collected. Screening of the APC and human mutY homolog (MUTYH) genes for germline mutations was conducted by direct polymerase chain reaction (PCR) sequencing. In two pedigrees, a heterozygous deletion in exon 16 of the APC gene was present in all FAP patients but absent in the unaffected individuals. There were no changes to the MUTYH gene. The first pedigree, with a new frameshift mutation at c.3926_3930 del AAAAG (p. Glu1309Aspfs X4), exhibited obvious differences in the polyp number such that the proband manifested only three colorectal polyps, whereas another patients showed the symptoms of classic FAP. The second pedigree, also traced a new mutation at c.3922_3925 del AAAG (p. Glu1309Argfs X11). Although all of the patients presented with classic polyposis, one of them exhibited a delayed onset of colorectal cancer in his 50s. Two novel mutations at codon 1309 in two Chinese families suffering from FAP could enrich the germline mutation spectrum of the APC gene. Families of individuals might manifest different phenotypes, even with an identical codon 1309 mutation, unlike in previous studies.

Microenvironment-evoked cell lineage conversion: Shifting the focus from internal reprogramming to external forcing.

Seeking possible ways to create replacement cells for the faded ones with deficits in functionality or quantity inspires comprehensive needs for cell lineage conversion. To fulfill this promise, reprogramming and microenvironment direction have been used to manipulate abundant cell fates. We briefly describe the evolution and fundamental insights of these two major strategies applied for lineage specification, comment generally on their current limitations, and analyze the orchestral interplay between them. We also present several future directions and discuss the potential clinical uses. Based on the relatively slight safety and technical issues, we conclude that microenvironment-evoked cell lineage conversion, instead of reprogramming, will be the shifting focus in regenerative medicine.

MSCs: Biological characteristics, clinical applications and their outstanding concerns.

Mesenchymal stem cells (MSCs) are multi-potent adult stem cells harboring multi-lineage differentiation potential and immunosuppressive properties that make MSCs an ideal candidate cell type for immunomodulation and regenerative medicine. Currently, MSC-related researches and clinical trials have evoked exciting promise in a variety of disorders and tissue regeneration. However, it must be recognized that several critical potential problems have also emerged from current clinical trials, for example: (1) the indefinite association between the phenotypic characteristics and the biological functions of MSCs; (2) the lack of clinical data to support the long-term safety of MSCs; (3) the need for further clarification of multiple mechanisms of MSC transplant actions in vivo; and (4) the lack of comparability of MSC transplant efficacy. Therefore, MSC-based therapies could not yet be considered a routine treatment in the clinic. Based on these, we proposed that large-scale and multi-center clinical trials of MSC-based therapies should be initiated under strict supervision. These interventions might help to establish a new clinical paradigm to turn MSC transplantation into a routine therapy for at least some diseases in the near future.

Prevalence of BK virus and JC virus in peripheral blood leukocytes and normal arterial walls in healthy individuals in China.

Several studies have demonstrated that BK virus (BKV) and JC virus (JCV) establish latent infection in peripheral blood leukocytes (PBLs) of healthy individuals; however, the main populations studied are European. In this study, the prevalence of BKV and JCV DNA in PBLs from healthy adult individuals and umbilical cord blood from newborn children in China was detected by semi-nested polymerase chain reaction (snPCR) followed by restriction enzyme analysis. The results suggest that the healthy adult Chinese population harbors BKV and JCV DNA in peripheral leukocytes. Overall, the prevalence of BKV and JCV DNA in PBLs of healthy adult individuals was 42.1% and 7.8%, respectively. The overall prevalence of BKV DNA was significantly higher than that of JCV DNA. None of the umbilical cord blood samples from newborn children were positive for BKV and JCV DNA. To understand further the target tissues involved in establishment of BKV and JCV latency in healthy individuals, the presence of DNA from both viruses was detected in normal arterial wall samples from 20 young trauma victims by the same method used for leukocyte DNA. BKV DNA was detected alone in 20% of samples tested; JCV DNA was not detected alone in any of the samples. DNA from both viruses was found in 5% of samples. This is the first report to show that normal arterial walls of healthy individuals may be another target site of latency for BKV and JCV.

[Inhibitory effect of antisense human telomerase reverse transcriptase(hTERT) on telomerase activity of human pulmonary giant cell carcinoma cell line(PLA-801D)].

BACKGROUND & OBJECTIVE: Telomerase has been thought to play an important role in the carcinogenesis in recent years. Human telomerase reverse transcriptase (hTERT) is a limiting component for telomerase activity. This study was designed to explore the effect of transfection of the full-length cDNA of antisense hTERT on the malignant phenotype of human pulmonary giant cell carcinoma cell line (PLA-801D) and its potential role in the gene therapy for cancers. METHODS: An antisense hTERT cDNA eukaryotic expression vector pcDNA3.1(-)-hTERT including the full length of hTERT cDNA sequence was constructed using recombinant DNA technique and transfected into human pulmonary giant cell carcinoma cells (PLA-801D) with liposome. The effect of antisense hTERT on the cellular proliferation capacity of PLA-801D cells was analyzed by the growth curve. The expression of hTERT mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR). The telomerase activity was determined by telomeric-repeat amplification protocol enzyme-linked immunoassay (TRAP-ELISA). RESULTS: Antisense pcDNA3.1 (-)-hTERT eukaryotic expression have been constructed and was successfully transfected into the PLA-801D cells. The growth speed of PLA-801D transfected with antisense hTERT was significantly inhibited compared with the control cells, and the hTERT mRNA expression was inhibited, the relatively expression was only 15.7% of control cells, and telomerase activity was down-regulated about 82.4%. CONCLUSION: Full-length antisense hTERT cDNA can suppress hTERT mRNA expression and telomerase activity, and restrict the growth speed of tumor cells.