Photocatalytic Three-Component Reaction for the Synthesis of Multifunctional Diaryl Sulfides.

A photocatalytic three-component reaction of a nitroarene, a thiophenol, and a ketone for the synthesis of multifunctional diaryl sulfides was reported using a nitro group as the nitrogen source and thiophenol as the sulfur source. Thiophenol also serves as a proton donor to reduce nitroarene to arylamine as a key intermediate for the formation of C-N and C-S bonds. Good functional group tolerance and mild reaction conditions make this method have practical synthetic value for diversified multifunctional diaryl sulfides.

Anti-hepatitis B virus activity and hepatoprotective effect of des(rhamnosyl) verbascoside from Lindernia ruellioides in vitro.

Although clinically approved hepatitis B virus (HBV) polymerase inhibitors (lamivudine-3TC, entecavir, etc.) serve as effective therapeutics, the virus can easily generate resistance to them. Therefore, the treatment of HBV infection remains a public health problem. Numerous studies have shown that natural products have prospective anti-HBV activity. The purpose of this study was to isolate and extract des(rhamnosyl) verbascoside from Lindernia ruellioides (Colsm.) Pennell and explore its anti-HBV and hepatoprotective effects. Anti-HBV activity was evaluated in HepG2.2.15 cells, a human hepatocellular carcinoma cell line with HBV-stable infection, and its protective effect was evaluated in HL-7702 cells, a normal human liver cell line. HepG2.2.15 cells maintained normal growth morphology within the selected concentration range of des(rhamnosyl) verbascoside. It also inhibited the expression of HBV antigens and HBV DNA in a dose- and time-dependent manner in vitro. Further, western blot experiments showed that it could downregulate HBV X protein (HBx) expression in a dose-dependent manner. In the H2 O2 -induced hepatocyte injury model, the cell-survival rate of the HL-7702 cells with the highest drug dose reached 85.25%, which was significantly improved compared with that of the model group. Most of the cells returned to normal morphology, showing polygonal or fusiform structures. Thus, it may be stated that des(rhamnosyl) verbascoside exhibits anti-HBV activity and hepatoprotective effects in vitro and may exert an anti-HBV effect via antigen inhibition, HBV DNA secretion, and HBx protein expression.

Developing a Web-Based Comic for Newly Diagnosed Women With Breast Cancer: An Action Research Approach.

BACKGROUND: Personal narratives have been seen as a useful way of communicating about cancer treatment options and providing recovery information. Many printed versions of such material are available, including comics that explore the individual memories of patients who have gone through cancer treatment. These studies have been used to orientate patients, patients' relatives, and physicians. However, only a few Web-based comics have been specifically designed for patients with breast cancer and used as aids to decision making. OBJECTIVE: We aimed to describe the developmental process of creating an animated comic as a Web-based surgery decision-making tool; the comic was aimed at illustrating the feelings, thoughts, and meanings when a patient suffers from breast cancer. This was done by recounting the symptoms, diagnostic process, treatments, and treatment effects of such women from the diagnosis stage onward. METHODS: Using cycles of planning, action, evaluation, and reflection, which involved collaborative work, action research was conducted to develop a Web-based animated comic. The stages of action research consisted of (1) semistructured and in-depth interviews to collect experiences of women with breast cancer; (2) construction of an animated comic by editors, graphics designers, dubbers, and information technology engineers; (3) redrawing of pictures of the comic after gathering feedback from a breast surgeon; and (4) evaluation of the Web-based animated comic using 6 patient focus groups. RESULTS: The comic was produced and showcased on the website "The Network of Making-decision Aids for Breast Cancer Surgery"; the comic was accompanied by soft music and audio explanations. The comic functions as a personal statement that describes experiencing breast cancer. The animated comic consists of 8 chapters, based on the 8 themes deducted from the findings obtained during the analysis of relevant interviews. The 8 chapters include (1) the appearance of a lump; (2) confirmation by medical diagnosis; (3) the uncertainty of waiting (4) fear of life-threatening disease; (5) choosing life over despair; (6) being brave and deciding to undergo treatment; (7) choosing the type of surgery; and (8) being reborn. CONCLUSIONS: Using action research, this study illustrated that the comic that sheds light on issues of feelings, emotions, and thoughts that are present when a woman is diagnosed with breast cancer and provides a communication medium to explain the steps in the process. Meanwhile, it implies that hope will be able to overcome the challenges that will be faced. Within the Web-based decision aid for patients with breast cancer, the animated comic acts as an information resource and is aimed at patients' understanding of impacts of emotions arising when suffering from breast cancer. It is potentially applicable as a therapeutic tool that facilitates self-reflection and self-healing among newly diagnosed patients with breast cancer.

[Serum levels of interleukin-38 and interleukin-1ß in the acute phase of Kawasaki disease in children].

OBJECTIVE: To study the expression of serum cytokines, interleukin-38 (IL-38) and interleukin-1ß (IL-1ß) in the acute phase of Kawasaki disease (KD) in children and the association of IL-38 and IL-1ß with inflammatory response in the acute phase and the development of coronary artery lesion (CAL). METHODS: A total of 40 children with KD who were hospitalized in the hospital between July 2015 and June 2016 were enrolled, with 21 children in the CAL group and 19 in the non-CAL (NCAL) group. Thirty healthy children and 19 children with infection and pyrexia, who were matched for sex and age, were enrolled as healthy control group and pyrexia control group respectively. ELISA was used to measure the serum levels of IL-38 and IL-1ß in the 40 children in the acute phase of KD. Spearman's rank correlation analysis was used to investigate the correlations of IL-1ß and IL-38 with interleukin-6 (IL-6), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), procalcitonin (PCT), N-terminal pro-brain natriuretic peptide (NT-proBNP), triglyceride (TG), and total cholesterol (TC). RESULTS: The serum level of IL-38 in the children in the acute phase of KD was significantly lower than that in the healthy control group (P<0.05), but significantly higher than that in the pyrexia control group (P<0.05). There was no significant difference in the level of IL-38 between the CAL and NCAL groups (P>0.05). The children in the acute phase of KD had a significantly higher level of IL-1ß than the healthy control group (P<0.05), while there was no significant difference between this group and the pyrexia control group (P>0.05). There was also no significant difference in the level of IL-1ß between the CAL and NCAL groups (P>0.05). Serum IL-1ß and IL-38 levels were not correlated with serum levels of CRP, ESR, PCT, IL-6, and NT-ProBNP or blood lipids (TG and TC) (P>0.05). CONCLUSIONS: IL-38 is involved in an inflammatory response in the acute phase of KD and may exert an anti-inflammatory effect, which is opposite to the effect of IL-1ß to promote inflammatory response. However, there is no significant correlation between these two cytokines and the development of CAL in KD.

Cross-talk between α7 nAChR-mediated cholinergic pathway and acylation stimulating protein signaling in 3T3-L1 adipocytes: role of NFκB and STAT3.

Inflammation is a key feature in adipose tissue, especially in association with obesity comorbidies. The novel adipokine acylation stimulating protein (ASP) is one factor implicated in the inflammatory response. The disruption of the α7 nicotine acetylcholine receptor (α7nAChR), an important component of the endogenous non-neural cholinergic defense system, may exacerbate sustained inflammatory phenotype. We examined cholinergic regulation of ASP-initiated inflammatory response in 3T3-L1 adipocytes. Our results show that preincubation of 3T3-L1 cells with α7nAChR agonist GTS-21 significantly reduces ASP-mediated chemokine MCP-1 secretion, which is regulated though nuclear factor κB (NFκB) and signal transducer and activator of transcription 3 (STAT3). Treatment of 3T3-L1 cells with GTS-21 significantly reduced NFκB activation by DNA binding and STAT3 activation by disturbing post-translational modification.

Elevated visfatin levels in obese children are related to proinflammatory factors.

BACKGROUND: Obesity and related metabolic diseases are associated with a state of chronic low-grade inflammation,which is characterized by abnormal cytokine production and increased synthesis of proinflammatory proteins.Recent studies have indicated that visfatin is both an adipokine and an inflammatory cytokine. OBJECTIVE: In this study, we assessed the association between serum visfatin concentrations and markers of systemic inflammation [high-sensitivity C-reactive protein (hs-CRP), interleukin 6 (IL-6), and tumor necrosis factor a (TNF-a)] and insulin resistance in Chinese obese children. METHODS: A total of 44 obese Chinese children (23 boys and 21 girls) and 50 age- and gender-matched normal weight children (23 boys and 27 girls) were enrolled in the study. Anthropometric measurements and blood pressure were taken. Fasting levels of visfatin, hs-CRP, IL-6, TNF-a,glucose, insulin, and lipid profile were assayed, and the homeostasis model assessment for insulin resistance was calculated as a marker of insulin resistance. RESULTS: Serum visfatin levels [obese group (OB):7.48±0.39 vs. C: 5.31±0.28, pO.OS); there were no significant differences in visfatin, hs-CRP, IL-6, and TNF-a concentrations between girls and boys. The correlations between visfatin and anthropometric, metabolic parameters,and inflammatory [body mass index (BMI), waist circumference, triglyceride, hs-CRP, IL-6, TNF-a] revealed differences between genders; visfatin correlated with BMI(r=0.247, p=0.029) and IL-6 (r=0.427, p=0.013) in boys only. CONCLUSION: The association of circulating visfatin with anthropometric parameters, metabolic parameters, and inflammatory factors is dependent on gender, although no such correlation was observed between serum visfatin concentration and insulin resistance. Visfatin could bean important inflammatory cytokine representing a low grade inflammatory state in obesity.

[Identification of a rare allele FGA-13 in Guangdong Han population].

OBJECTIVE: To report on a rare allele FGA-13 identified in Guangdong Han population. METHODS: The rare allele was detected by PCR-STR and DNA sequencing. RESULTS: The core repeat sequence of rare allele FGA-OL is [TTTC]3[TTTT][TTCT][CTTT]5 [CTCC][TTCC]2, which has been determined as FGA-13. CONCLUSION: The rare allele FGA-13 is also present in Guangdong Han population. This is significant for personal identification and paternity testing.

[Detection of matrix metalloproteinase-11 in menstrual blood by enhanced chemiluminescence method].

OBJECTIVE: To explore the forensic application value of detection of matrix metalloproteinase-11 (MMP-11) in menstrual blood by enhanced chemiluminescence method. METHODS: Menstrual blood, vaginal swab, peripheral blood, saliva stain, urine stain and semen stain were collected to detect whether or not there were MMP-11 using enhanced chemiluminescence method. The specificity and reliability of the MMP-11 assay along with its sensitivity were evaluated. RESULTS: The positive detection rate of MMP-11 in menstrual blood was 89.47%, whereas no MMP-11 was found in vaginal swab, peripheral blood, saliva stain, urine stain and semen stain. When 25 microL sample was added, the mass concentration of protein was 1.329 microg/microL, then MMP-11 could be detected. A positive detection rate of 89.58% was observed in MMP-11 positive menstrual blood samples after stored at 4 degrees C for 20 months. CONCLUSION: Enhanced chemiluminescence method is sensitive and specific for detecting MMP-11, and can be applied to distinguish menstrual blood from common stain such as peripheral blood, vaginal fluid.

[Identification of sibling brothers using STR and Y-biallelic markers].

OBJECTIVE: To explore the methods for identification of sibling brothers with Y-STR locus mutation by detection of genetic markers on autosome and Y-biallelic. METHODS: Goldeneye 20A and 18NC kit were used to genotyped the 35 STRs on autosome from two men. PowerPlex Y kit and Yfiler kit were used to genotyped the 16 STRs on Y chromosome full sibling index was calculated by ITO method. Twenty Y-biallelic markers were genotyped by fragment length discrepant allele specific PCR or general PCR. RESULTS: Relationship of sibling brothers was found to have mutation of 2 loci on 16 Y-STR and the identical genetype of 20 Y-biallelic markers as well as a cumulative full sibling index of 4.3149 x 10(6) from 35 STRs on autosome. CONCLUSION: In identification of paternal linage of Y-STR mutation, more genetic information can be acquired by detection of Y-biallelic markers including SNP and InDel.

Photocatalytic Synthesis of Sulfinamides and Sulfoxides from Nitroarenes and Thiophenols.

We present an efficient and versatile visible light-driven methodology for synthesizing sulfinamides and sulfoxides using nitroarenes as the nitrogen source and thiophenols as the sulfur source. The switch-over of the two reaction pathways was achieved by changing the type of photocatalyst and the amount of thiophenol in the reaction mixture. The reaction proceeds under mild conditions with good functional group tolerance and can easily be scaled up.

The molecular mechanism of acylation stimulating protein regulation of adipophilin and perilipin expression: involvement of phosphoinositide 3-kinase and phospholipase C.

The novel adipokine acylation stimulating protein (ASP) is involved in lipid metabolism and obesity-related disorders. Adipophilin and perilipin, two members of the lipid droplet protein family, participate not only in fat storage within adipocytes, but also in ectopic lipid deposition in the form of cytoplasmic triglyceride (TG) droplets within many types of mammalian cells. During differentiation to mature adipocytes, mechanisms controlling the synthesis and turnover of these lipid droplet proteins are only partially understood, the mechanisms regulating gene/protein expression as yet unidentified. In our previous study, ASP has been shown to regulate adipophilin and perilipin expression to facilitate TG synthesis during 3T3-L1 cell differentiation. Our aim in this study was to provide insight into the physiological importance of phosphoinositide 3-kinase (PI3K) and phospholipase C (PLC) in ASP-triggered alteration of adipophilin and perilipin expression. We found that acute (2.5 h) inhibition of PLC or PI3K results in a decrease in mRNA and protein of perilipin and adipophilin at any time during differentiation. The fact that there is such a rapid change even with mRNA levels suggests a rapid turnover of both mRNA and protein independent of a direct ASP effect. Also, the presence of these inhibitors blocked the ASP stimulatory effects with a maximal decrease in gene and protein expression of adipophilin (-45% and -60%, respectively, P < 0.01) and perilipin (-96% and -63%, respectively, P < 0.01 and P < 0.05). These findings provide further understanding of the adipogenic properties of ASP in adipocytes.

[Calculation of the avuncular index].

OBJECTIVE: To introduce the method of avuncular index (AI) calculation. METHODS: Identity by decent coefficient, coancestry coefficient and AI law were employed in identification of uncle-niece relationship, when autosomal STR loci were detected to determine controversial uncle-niece relationship. RESULTS: The results of AI calculation were coincidental using identity by descent coefficien, coancestry coefficient and AI law. CONCLUSION: The results are coincidental using three methods in the different situations. AI index is higher with participation of children's mother.

Biochanin A Alleviates Cerebral Ischemia/Reperfusion Injury by Suppressing Endoplasmic Reticulum Stress-Induced Apoptosis and p38MAPK Signaling Pathway In Vivo and In Vitro.

We have previously shown that biochanin A exhibits neuroprotective properties in the context of cerebral ischemia/reperfusion (I/R) injury. The mechanistic basis for such properties, however, remains poorly understood. This study was therefore designed to explore the manner whereby biochanin A controls endoplasmic reticulum (ER) stress, apoptosis, and inflammation within fetal rat primary cortical neurons in response to oxygen-glucose deprivation/reoxygenation (OGD/R) injury, and in a rat model of middle cerebral artery occlusion and reperfusion (MCAO/R) injury. For the OGD/R in vitro model system, cells were evaluated after a 2 h OGD following a 24 h reoxygenation period, whereas in vivo neurological deficits were evaluated following 2 h of ischemia and 24 h of reperfusion. The expression of proteins associated with apoptosis, ER stress (ERS), and p38 MAPK phosphorylation was evaluated in these samples. Rats treated with biochanin A exhibited reduced neurological deficits relative to control rats following MCAO/R injury. Additionally, GRP78 and CHOP levels rose following I/R modeling both in vitro and in vivo, whereas biochanin A treatment was associated with reductions in CHOP levels but further increases in GRP78 levels. In addition, OGD/R or MCAO/R were associated with markedly enhanced p38 MAPK phosphorylation that was alleviated by biochanin A treatment. Similarly, OGD/R or MCAO/R injury resulted in increases in caspase-3, caspase-12, and Bax levels as well as decreases in Bcl-2 levels, whereas biochanin A treatment was sufficient to reverse these phenotypes. Together, these findings thus demonstrate that biochanin A can alleviate cerebral I/R-induced damage at least in part via suppressing apoptosis, ER stress, and p38 MAPK signaling, thereby serving as a potent neuroprotective agent.

[Effect of acupuncture on pain and cerebral hemodynamics in patients with migraine: a randomized controlled trial].

OBJECTIVE: To observe the therapeutic effect between acupuncture combined with medication and simple medication on migraine and cerebral hemodynamics. METHODS: A total of 120 patients with migraine were randomized into an acupuncture plus medication group (60 cases, 3 cases dropped off) and a medication group (60 cases, 6 cases dropped off). In the medication group, flunarizine hydrochloride capsule was given orally before sleep, 10 mg a day. On the basis of the treatment in the medication group, acupuncture was applied at Sizhukong (TE 23), Shuaigu (GB 8), Taiyang (EX-HN 5), Fengchi (GB 20) and etc. in the acupuncture plus medication group, 30 min each time, once a day. Treatment for 4 weeks was required in both groups. Before and after treatment, the visual analogue scale (VAS) score, indexes of cerebral hemodynamic [blood flow velocity of anterior cerebral artery (ACA), middle cerebral artery (MCA), posterior cerebral artery (PCA), vertebral artery (VA) and basilar artery (BA)] and total TCM syndrome score were observed, and the clinical therapeutic effect and the incidence of the adverse events were evaluated in both groups. RESULTS: Compared before treatment, the VAS scores, the blood flow velocity of ACA, MCA, PCA, VA, BA and the total TCM syndrome scores were decreased in both groups (P<0.05). After treatment, the VAS score, the blood flow velocity of ACA, MCA, PCA, VA, BA and the total TCM syndrome score in the acupuncture plus medication group were lower than those in the medication group (P<0.05). The total effective rate was 96.5% (55/57) in the acupuncture plus medication group, which was superior to 90.7% (49/54) in the medication group (P<0.05). There was no statistical difference in the incidence of adverse events between the two groups (P>0.05). CONCLUSION: Acupuncture combined with flunarizine hydrochloride capsule can effectively relieve the pain in patients with migraine, reduce the cerebral blood flow velocity, the efficacy is superior to simple flunarizine hydrochloride capsule.

Polymorphism analysis and evaluation of nine non-CODIS STR loci in the Han population of Southern China.

BACKGROUND: Knowledge of allele and genotype frequencies is an essential prerequisite to the use of any human polymorphism in forensic work. AIM: To study the genetic polymorphism and evaluate the application value of nine STR loci. SUBJECTS AND METHODS: Genotyping of nine STR loci, including D11S2368, D12S391, D13S325, D18S1364, D22-GATA198B05, D6S1043, D2S1772, D7S3048 and D8S1132, of 1050 unrelated individuals was performed with the STR_Typer_10_v1 kit and Genetic Analyzer 3100 and analyzed with PowerState V12.xls and Arlequin ver 3.11 analyzing software. RESULTS: Allele frequency distribution was statistically analyzed and Hardy-Weinberg equilibrium determined. Several common parameters used in forensic sciences were found: the heterozygosity (H) ranged from 0.827 to 0.892; the matching probability (MP) ranged from 0.029 to 0.074; the power of discrimination (PD) ranged from 0.926 to 0.971; the power of exclusion (PE) ranged from 0.649 to 0.779; the polymorphic information content (PIC) ranged from 0.77 to 0.86; and the typical paternity index (TPI) ranged from 2.88 to 4.62. CONCLUSION: The results indicate that nine STR loci are high polymorphic among the Han population in Southern China. This set of polymorphic STR loci is a useful tool in forensic paternity testing and anthropological study.

[Effect of acylation stimulating protein on the perilipin and adipophilin expression during 3T3-L1 preadipocyte differentiation.].

Perilipin and adipophilin, two significant lipid droplet (LD)-specific proteins, participate in storing fat or ectopic lipid deposition and fat mobilization in many types of mammalian cells. Acylation stimulating protein (ASP) is a novel adipocyte-derived hormone known for a major determinant for triglyceride synthesis (TGS) and lipid metabolism. The present study was aimed to investigate: (1) whether ASP, rather than insulin, is a powerful potentiator which could physiologically and directly influence TGS during 3T3-L1 preadipocyte differentiation; (2) whether ASP exposure at indicated time points during 3T3-L1 preadipocyte differentiation could influence the gene/protein expression of adipophilin and perilipin. 3T3-L1 preadipocytes were differentiated by traditional hormone cocktail and divided into control, ASP and insulin groups according to the treatment of ASP (1 mmol/L) or insulin (100 nmol/L). ASP-stimulated and insulin-stimulated TGS rate at indicated time points (0 d, 3 d, 6 d, 9 d) were assayed by measuring the incorporation of [(3)H]-oleic acid into TG, and the corresponding glucose transport was assayed by [(3)H]-2-DG uptake. The effects of ASP or insulin on gene/protein expression of adipophilin and perilipin at indicated time points were evaluated by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The results obtained were as follows: (1) on the 3rd and 6th day of differentiation, ASP dramatically enhanced TGS rate compared with control group (P<0.05, P<0.01); There was no significant difference in TGS rate between insulin group and control group; (2) on the 6th and 9th day of differentiation, both ASP and insulin promoted glucose uptake (P<0.05, P<0.01), and the promoting effect in ASP group was greater than that in insulin group; (3) ASP elevated adipophilin gene and protein expression at the very early stage of differentiation (P<0.05, P<0.001) and had no significant effect from the 4th day of differentiation. Perilipin gene and protein expression increased throughout preadipocyte differentiation and its expression was up-regulated following ASP stimulation from the 3rd day of differentiation (P<0.05, P<0.001) to the end of differentiation (P<0.05); (4) Insulin did not affect gene and protein variation pattern of adipophilin and perilipin. Taken together, this study provides evidence that ASP-evoked changes in gene and protein expression of adipophilin and perilipin correlate with ASP-stimulated TGS acceleration, and adipophilin and perilipin are involved in the molecular mechanism of ASP-induced adipogenesis and LD formation.

[A new method for calculating paternity index].

OBJECTIVE: To introduce a new method for calculating the paternity index (PI). METHODS: Assuming that each allele from parents has undergone a transition before it segregates and transmits to child. The transition probability is 1 when parent allele is the same as child's, the transition probability is 0 when parent allele is different from the child's. Every allele has a transmission probability with 0.5. Base on these theories, it is easy to gain the probability that child inherits an allele from the alleged father or mother. Thus, the X value (numerator) and Y value (denominator) of PI formula can be calculated, as unknown man provide an allele for child with the allele frequency. RESULTS: A general formula that calculated the PI for trios, duos and missing child cases was deduced. CONCLUSION: The new method is practical in all kinds of forensic paternity case.

[A new method for calculating the paternity index in presence of mutation].

OBJECTIVE: To introduce a method of calculating the paternity index (PI) for autosomal codominant markers when the mutation is encountered. METHODS: Assuming mutation arises with a mutational probability before an allele segregates and transmits to child with 1/2 chance, the probability of a random man giving an allele to child is the allele frequency in population. Considering only one mutation event in one case, the mutant allele can be determined by comparing the allele between parents and child. Furthermore, the probability of child's genotype can be calculated to identify the father between a tested man and a random male. Subsequently, the PI can be calculated. RESULTS: Formula of PI was deduced for trios, duos and missing child cases, including maternal mutation. CONCLUSION: The method is easy to understand and useful in paternity testing.

[Application of ITO method and discriminant functions in full sibling and half sibling identification].

OBJECTIVE: To investigate the application of ITO method and discriminant functions method in full sibling and half sibling identification. METHODS: Five hundred pairs of full siblings (FS), 50 pairs of half siblings (HS) and 500 pairs of unrelated individuals (UR) were genotyped by PowerPlex 16 system. Full sibling index (FSI), half sibling index (HSI) and the FSI:HSI ratio were calculated with ITO method. Allelic matching of each pair of the three groups was compared. The locus numbers of no-allele sharing (x0), half-allele sharing (x1) and two-alleles sharing (x2) were calculated, respectively. The discriminant functions about full-siblings, half-siblings and unrelated individuals (UR) were established by SPSS 13.0 statistical software. RESULTS: (1) Regard FSI > or = 19 or FSI < 1 as the standard of distinguishing full sibling from unrelated individual, the alternate correct percentage was 96.4%. Regard HSI > or = 19 or HSI < 1 as the standard of distinguishing half sibling from unrelated individual, the alternate correct percentage was 85.3%. Regard FSI:HSI > or = 1 or FSI:HSI < 1 as the standard of distinguishing full sibling from half sibling, the alternate correct percentage was 87.5%. (2) Four groups of discriminant functions were established. The alternate correct percentage of these discriminant functions were 84.4%-97.7%, with the highest one in full sibship-unrelated individual group. CONCLUSION: Both ITO method and discriminant functions method are efficient in identification of full sibling or half sibling.

[Effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocytes and its possible mechanisms].

OBJECTIVE: To investigate the effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocyte, and study the possible mechanisms. METHODS: 3T3-L1 preadipocytes were cultured in vitro. The proliferation potentials of 3T3-L1 preadipocytes that were treated with different concentrations of ghrelin were evaluated by MTT methods. The levels of c-myc and thymidine kinase mRNA were detected using RT-PCR. 3T3-L1 preadipocytes were differentiated into the matured adipocytes with insulin (INS) or ghrelin. The morphological changes of 3T3-L1 adipocytes were observed and the differentiation rate was assayed by oil-red O staining. Total RNA was extracted from adipocytes at various times, and the levels of peroxisome proliferation activated receptor gamma (PPARgamma) and CAAT/enhancer binding protein(C/EBPalpha) mRNA expressions were detected using RT-PCR. RESULTS: Ghrelin at concentrations of 10(-7) to 10(-15) mol/L significantly stimulated preadipocyte proliferation (p<0.05). The levels of c-myc and thymidine kinase mRNA significantly increased in 3T3-L1 preadipocytes with 10(-9) mol/L and 10(-11) mol/L ghrelin treatment (p<0.01). The 3T3-L1 preadipocytes treated with 10(-11) mol/L ghrelin had lots of lipid droplets in the cytoplasma, but the differentiation rate was lower than those treated with INS. Ghrelin of 10(-11) mol/L significantly increased the mRNA expression of PPARgamma and C/EBPalpha in the course of 3T3-L1 preadipocyte differentiation, compared with the normal control group (p<0.05). The PPARgamma and C/EBPalpha mRNA expression increased with the prolonged differentiation of preadipocytes induced by ghrelin or INS. There were significant differences in the levels of PPARgamma and C/EBPalpha mRNA expression between the 2nd and 8th days of differentiation(p<0.01). CONCLUSIONS: Ghrelin promotes the proliferation and differentiation of 3T3-L1 preadipocytes. The proliferation of 3T3-L1 preadipocytes induced by ghrelin may be associated with increased c-myc levels. Ghrelin may promote differentiation of 3T3-L1 preadipocytes by increasing mRNA expression of PPARgamma and C/EBPalpha, thus enhances the sensitivity of adipocytes to INS.