[Progress and application of liver organoids in the study of liver cancer].

The incidence rate of liver cancer has been rising in recent years. Traditional cell line culture and human patient-derived tumor xenograft models, which are commonly used tools to simulate the occurrence of human liver cancer, have deepened the understanding of tumor occurrence, development, and drug resistance mechanisms. However, they cannot reflect the accurate state of cancer cells, the tumor microenvironment, or spatial structural characteristics. Recently, more in vitro-produced physiological liver organoids have been applied in the study of liver cancer. Liver organoid models have made breakthroughs in the occurrence and development mechanisms of liver cancer, personalized drug screening and biomarker identification, immunotherapy, and regenerative medicine applications. This paper mainly summarizes the progress and application of liver organoids processed in the study of liver cancer.

Effect of age at first egg on reproduction performance and characterization of the hypothalamo-pituitary-gonadal axis in chickens.

As an important indicator of sexual maturity of hens, age at first egg (AFE) is significantly associated with reproduction performance. In this study, 400 hens were divided into 6 groups based on AFE to analyze the difference of reproduction performance, reproduction hormone levels and the characterization of the hypothalamo-pituitary-gonadal axis in chickens. The results showed that the egg production of the hens in the late-maturing groups was significantly lower than that of the ones in other groups and the precocious hens had a lower egg production rate. The hens in late-maturing group had a lower fertility rate, LH levels and shorter duration of peak of egg production (PEP), the precocious hens had lower PRL levels. In addition, the characterization of the hypothalamo-pituitary-gonadal axis showed that the individuals with normal AFE had higher GNRH, GNRHR, ESR1, KITLG, and CYP11A1 expression levels than late-maturing and precocious individuals, which indicated that the chickens with normal AFE advantages on reproduction regulation system.

RFamide-related peptides' gene expression, polymorphism, and their association with reproductive traits in chickens.

RFamide-related peptides (RFRP) are synthesized by the hypothalamus and have a regulatory role in gonad development. The goal of this study was to investigate the association between SNP of the RFRP gene and the reproductive traits and hormone levels of Zhenning yellow chickens. The mRNA expression levels were detected based on different tissues, ages, and genotypes. Eleven mutation sites were detected in the RFRP gene, 4 of which were significantly related to reproductive traits and hormone levels. Association analysis revealed that A276G was associated with egg production at 300 d of age (EP300) and amount of prehierarchical follicles (P < 0.05). G1396A was associated with egg weight at 300 d of age and luteinizing hormone (LH) and prolactin levels (P < 0.05). G1694A showed significant associations with fertilization rate and LH levels (P < 0.05), and A2659G was associated with EP300 (P < 0.05). The results of expression analysis showed that the RFRP mRNA expression levels in the hypothalamus were higher than those in other tissues (P < 0.01). The expression in immature individuals was higher than that in mature ones (P < 0.01). There were also differences in mRNA expression levels between different genotypes (P < 0.05). In summary, the results of this study might provide potential markers and a theoretical basis for the improvement of chicken reproductive traits.

H-FABP gene expression and genetic association with meat quality traits in domestic pigeons (Columba livia).

1. H-FABP (heart-type fatty acid-binding protein) is an important fatty acid-binding protein that participates in the metabolism and intracellular transportation of lipids. This study was designed to study the H-FABP gene expression and genetic association with meat quality traits in domestic pigeons.2. The spatio-temporal expression patterns showed that H-FABP was widely expressed in all eleven tissues in birds from hatching to 4 weeks of age, and the expression level in the liver was the highest, and spleen and subcutaneous fat showed relatively high levels at all time points, and increased markedly as squabs became older.3. Nine novel SNPs were found; three of them (g.42 C > T and g.123 C > T in exon 1 and g.3128 C > T in exon 2) were synonymous mutations, and six of them (g.3304 T > C, g.3305 G > A, g.3313A>G, g.3343 C > T, g.3372A>C, g.3410 T > C in exon 3) were located in the 3' untranslated region (UTR). In addition, four haplotypes and six diplotypes were formed and showed weak linkage disequilibrium. Association analysis showed that in the SNP of g.3305 G > A, the AB genotype showed higher (P < 0.05) inosinic acid concentrations in breast muscle than the BB genotype. In the SNP of g.3313A>G, the AA and AB genotypes showed significantly higher (P < 0.01) inosinic acid concentrations than the BB genotype, AA showed the highest intramuscular fat content, and the relative mRNA expression level of the AA genotype was the highest (P < 0.05).4. Association analysis of diplotypes showed no correlation with meat quality traits. Moreover, correlation analysis indicated that the H-FABP mRNA expression level was tightly related to intramuscular fat content (P < 0.01).5. This study suggested that the H-FABP gene may be a candidate gene in marker-assisted selection of pigeons for high-quality meat.

MiR-129 reduces CDDP resistance in gastric cancer cells by inhibiting MAPK3.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-129 reduces CDDP resistance in gastric cancer cells by inhibiting MAPK3, by H.-Y. Cao, C.-H. Xiao, H.-J. Lu, H.-Z. Yu, H. Hong, C.-Y. Guo, J.-F. Yuan, published in Eur Rev Med Pharmacol Sci 2019; 23 (15): 6478-6485-DOI: 10.26355/eurrev_201908_18531-PMID: 31378887" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18531.

POMC gene expression, polymorphism, and the association with reproduction traits in chickens.

Reproduction trait is one of the most important economic traits in poultry industry. This study was aimed to investigate the mRNA expression levels, single nucleotide polymorphisms (SNP) of POMC gene, and the association with reproduction traits in chickens. Five SNP (g.958 G > A, g.1374 G > C, g.1393 G > A, g.1817 C > T, and g.1918G > A) were detected in introns of POMC gene in 317 Zhenning yellow chickens. Association analysis revealed that g.958 G > A and g.1817 C > T showed significantly associations with fertilization rate, hatching rate of hatching eggs, and hatching rate of fertilized eggs in chickens. Simultaneously, g.1374 G > C and g.1918G > A were both associated with egg weight at 300 D of age (P < 0.05). The SNP of g.958 G > A, g.1393 G > A, and g.1817 C > T were all associated with E2 hormone levels (P < 0.05). The result of mRNA expression levels in different tissues showed that POMC mRNA expression level in the pituitary was higher than those in the other tissues and varied in different genotypes. In conclusion, the results in this study provided new evidences that polymorphisms of the POMC gene have potential effects on reproduction traits in chickens. The 5 SNP detected in this study could be potential markers for improving reproduction traits in chickens.

MiR-129 reduces CDDP resistance in gastric cancer cells by inhibiting MAPK3.

OBJECTIVE: Abnormal expression of mitogen-activated protein kinase 3 (MAPK3) is related to invasion, metastasis, and drug resistance of multiple tumor cells. MiR-129 expression is associated with gastric cancer. Bioinformatics analysis showed a targeting relation between miR-129 and MAPK3. This study investigated whether miR-129 plays a role in regulating MAPK3 expression, affecting proliferation, apoptosis, and cisplatin (CDDP) resistance of gastric cancer cells. MATERIALS AND METHODS: The dual-luciferase reporter gene assay was used to assess the targeted regulation between miR-129 and MAPK3. The expression of miR-129 and MAPK3 in CDDP-resistant cell line MGC-803/CDDP and the parental MGC-803 cells was measured. MGC-803/CDDP cells were cultured in vitro and divided into miR-NC group and miR-129 mimic group. The expression of MAPK3 and p-MAPK3 protein were detected by Western blot and the effect of CDDP treatment on cell apoptosis and proliferation was detected by flow cytometry. RESULTS: There was a targeted regulation relation between miR-129 and MAPK3 mRNA. MiR-129 expression in MGC-803/CDDP cells was significantly lower than that in MGC-803 cells and the expression of MAPK3 mRNA and protein was significantly higher than that in MGC-803 cells. Compared with miR-NC group, the expression of MAPK3 and p-MAPK3 in MHC-803/CDDP cells in miR-129 mimic transfection group was significantly decreased, with increased cell apoptosis and reduced cell proliferation. CONCLUSIONS: The decreased expression of miR-129 and the up-regulation of MAPK3 are associated with CDDP resistance in gastric cancer cells. Overexpression of miR-129 inhibits MAPK3 expression and cell proliferation, it induces cell apoptosis and reduces CDDP resistance.

Expression of MSTN gene and its correlation with pectoralis muscle fiber traits in the domestic pigeons (Columba livia).

Myostatin (MSTN) is a negative regulator of skeletal muscle growth and plays an important role in muscle development. In this research, we constructed a tissue expression profile of the pigeon MSTN gene in eight tissues and a spatio-temporal expression profile in the pectoral muscle using qRT-PCR method. And the pectoralis muscle fiber traits during pigeon post-hatching stages at 1, 7, 14, 21, and 28 D were analyzed through the paraffin sections. Then the correlations between the muscle fiber diameter, cross-sectional area, density, and the expression of MSTN in the pectoral muscle were analyzed. Results showed that MSTN mRNA was mainly expressed in breast muscle, heart, spleen, and kidney and it was almost unexpressed in the liver and lungs. Moreover, the MSTN mRNA expression level in breast muscle was significantly higher than that in other tissues (P < 0.05), and showed an interesting trend that it decreased in the first week and then increased with age. Meanwhile, decrease of myostatin transcripts was accompanied by the down-regulation of Myf5 and the up-regulation of MyoG during the first week post-hatching. In addition, the paraffin sections analysis results revealed that the diameter and cross-sectional area of pectoralis muscle fiber significantly increased with age (P < 0.05), and a significant positive correlation was shown between the MSTN gene expression level and muscle fiber diameter (P < 0.05). These fundamental researches might contribute to further understanding of the roles MSTN played in the post-hatching muscle fiber development in pigeon.

NGS-based profiling reveals a critical contributing role of somatic D-loop mtDNA mutations in HBV-related hepatocarcinogenesis.

BACKGROUND: Somatic mutations of mitochondrial DNA (mtDNA) have been extensively identified mainly by traditional Sanger sequencing technology in various cancer types. However, low detection sensitivity of traditional methods greatly limits the comprehensive profiling of mtDNA somatic mutations in cancers, especially in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Moreover, the functional roles of mtDNA mutation in HBV-related hepatocarcinogenesis have not been systematically revealed. PATIENTS AND METHODS: Next-generation sequencing (NGS) platform was applied to profile the somatic mtDNA mutations of HCC and paired paratumor (non-HCC) tissues from a large cohort of 156 HBV-HCC patients. RESULTS: Our data revealed the common existence of mtDNA mutation in both inflammatory and cancer tissues with significantly different mutation pattern. The mutation density (mutation number/region length) of D-loop region was much higher than that of other regions in both HCC and non-HCC tissues. Unexpectedly, the average mutation number in D-loop region of HCC tissues was significantly less than that of non-HCC tissues. In contrast, the heteroplasmy level of D-loop region mutations was significantly increased in HCC tissues, implying that the D-loop mutations might be positively selected in HCC tissues. Furthermore, our results indicated that the patients with D-loop mutations had a significantly lower mtDNA copy number and were more likely to relapse. In vitro experiments demonstrated that proliferation, invasion and metastasis ability of HCC cells with D-loop region mutations were significantly higher than those without D-loop region mutations. CONCLUSION: These results emphasize the critical contributing role of somatic mtDNA D-loop mutations in HBV-related hepatocarcinogenesis.

Association of ADSL gene polymorphisms with meat quality and carcass traits in domestic pigeons (Columba livia).

1. Adenylosuccinate lyase (ADSL) plays an important role in the synthesis of inosine monophosphate (IMP). In this trial, a total of 200 pigeons were sampled and slaughtered. Seven meat quality traits and 11 carcass traits were measured. DNA sequencing was used to detect nucleotide mutations, and associations between ADSL gene polymorphisms and meat quality and carcass traits were analysed. 2. Sequencing results showed that 9 nucleotide mutations were found in the exons of the ADSL gene. All the mutations were synonymous except C13065G, which caused a change in amino acids (Ser to Arg). In addition, two of the detected single nucleotide polymorphisms (SNPs) had significant associations with meat quality and carcass traits. 3. For the C13065G SNP located in exon11, the IMP content of breast muscle in AA and AB genotype was higher than in the BB genotype (P < 0.01). The SNP G10696A located in exon10 was significantly associated with carcass rate, in which AA and AB genotype were higher than the BB genotype (P < 0.01). 4. The results indicated that the ADSL gene has a close association with meat quality and carcass traits in domestic pigeons, and G10696A and C13065G SNPs could be applied as genetic markers in molecular marker-assisted breeding of pigeons.

Effects of rearing systems on reproductive hormones secretion and their receptors gene expression in Xianju chickens under summer conditions.

Previous study in our lab showed that indigenous Xianju chickens from free-range system (FRS) under summer conditions had lower egg production than those from conventional cage rearing system (CRS). The objective of this study was to preliminarily determine the FRS-dependent mechanism of depressing laying performance according to determining the effect of rearing systems on reproductive hormones secretion and their receptors mRNA expression in Xianju chickens reared under summer conditions. A total of 360 indigenous Xianju chickens were randomly allocated to CRS and FRS groups, each of which included 5 replicates of 36 hens. The experiment lasted between 21 and 29 wk of age. We found that the ovarian weight, numbers of small yellow follicles, and large white follicles in the FRS group were lower than those in the CRS group (P < 0.05). Changing from CRS to FRS increased serum concentrations of prolactin and decreased serum-luteinizing hormone and progesterone levels (P < 0.05). Gene expressions in the preovulatory follicles from FRS hens were upregulated for prolactin receptor and downregulated for luteinizing hormone receptor and progesterone receptor, compared to those from CRS hens (P < 0.05). It can be concluded that changing from CRS to FRS in the current experimental conditions depressed egg production traits in Xianju chickens by inducing a synergistic activity of reproductive hormones and the gene expressions of their receptors.

[Treatment and prognosis of 264 patients with hypopharyngeal carcinoma].

Objective: To investigate the key factors influencing the prognosis of hypopharyngeal carcinoma and the therapeutic methods improving the efficacy of treatments for hypopharyngeal carcinoma. Methods: Two hundred and sixty-four cases of hypopharyngeal squamous cell carcinoma treated from May 2010 to May 2015 were analyzed retrospectively. There were 211 cases of pyriform sinus carcinoma, 37 cases of posterior pharyngeal wall carcinoma, and 16 cases of postcricoid carcinoma. According to UICC 2002 criteria, 2 cases were for stage Ⅰ, 14 for stage Ⅱ, 32 for stage Ⅲ and 216 for stage Ⅳ. Postoperative circumferential defects existed in 112 (42.4%) cases, and 86 of them were reconstructed with free jejunum transplantation. Among all cases, 54 patients (20.5%) had the preservation of laryngeal functions after surgery and 210 patients (79.5%) with total laryngectomy; 238 cases (90.2%) underwent bilateral cervical lymph node dissection and 203 patients received posterior pharyngeal lymph node exploration and dissection, with positive metastases for posterior pharyngeal lymph nodes in 36 cases (17.7%). Eight cases with cervical lymph node metastasis extensively involving the soft tissue, prevertebral fascia or encases carotid artery received preoperative radiotherapy of 50 Gy. After surgery 13 patients received concurrent radiotherapy and chemotherapy, 337 underwent adjuvant radiotherapy with a dose of 50-60 Gy each, and 14 patients did not receive radiotherapy or did not completed their radiotherapy programs. SPSS 13.0 saftware was used to analyze the data. Results: All patients were followed up for more than 2 years. With Kaplan-Meier method, the 2-, 3- and 5-years survival rates were 69.6%, 62.8% and 51.3%, respectively. There were significant differences in 3-year survival rates between T1-2 group (75.5%) and T3-4 group (59.2%) (χ(2)=4.282 P=0.039), N0 group (81.6%) and N+ group (58.2%) (χ(2)=6.802 P=0.009), laryngeal functions preserved (81.8%) and unpreserved group (58.9%) (χ(2)=5.314 P=0.021). Multivariate Logistic regression analysis showed that cervical lymph node metastasis was an independent prognostic factor (P=0.027). The success rate of free jejunum transplantation was 98.8%. Local recurrence, cervical lymph node recurrence, second primary cancer, and distant metastasis accounted respectively for 11.2%, 18.8%, 12.5% and 45.0% of death cases. Conclusions: The prognosis-associated factors for hypopharyngeal carcinoma should be taken into account, including the evaluation of the carcinogenesis of the mucosal area, early screening of premalignant lesion or second primary cancer in the esophagus and dissection of the posterior pharyngeal lymph nodes, which will help to improve the local control rate and recent survival rate in patients with hypopharyngeal cancer.

[Significance of retropharyngeal node dissection in treatment of hypopharyngeal carcinoma].

Objective: investigate the incidence of retropharyngeal lymph node (RPLN) metastasis and the risk factors for RPLN metastasis in hypopharyngeal cancer, and the relationship of planned dissection of the RPLN with the survival and tumor control rates in patients with hypopharyngeal cancer. Methods: A total of 203 patients with hypopharyngeal squamous cell carcinoma who underwent radical surgery as initial treatment from February 2011 to July 2015 were analyzed retrospectively. There were 167 cases of pyriform sinus carcinoma, 23 cases of posterior pharyngeal wall carcinoma, and 13 cases of postcricoid carcinoma. Results: The incidence of RPLN metastasis in HPC was 17.7%, with a highest rate of 43.5% in pharyngeal wall carcinoma. The incidence of RPLN metastasis in T3-4 pyriform sinus carcinoma was 18.3%, which significantly higher than 2.8% in T1-2 cases(χ(2)=5.360, P=0.020). The rate of RPLN metastasis was 23.8% in N2b-3 and 8.6% in N0-2a, with a statistically significant difference(χ(2)=7.637, P=0.006). There was no statistically significant difference in overall survival rates between patients with and without RPLN metastasis(P>0.05). Data were analyzed by SPSS 13.0 software. Conclusions: RPLN metastasis is not rare in hypopharyngeal carcinoma. Planned dissection of the RPLN should be performed with the initial surgery in patients with advanced hypopharyngeal cancer, especially posterior pharyngeal wall carcinoma, T3-4 pyriform sinus carcinoma and staged N2b-3 disease, which can reduce the regional recurrence rate and provided with a better prognosis.

[Endoscopic screening for upper gastrointestinal second primary malignancies in patients with hypopharyngeal squamous cell carcinoma].

Objective: To evaluate the usefullness of flexible esophagoscopy and chromoendoscopy with Lugol's solution in the detection of synchronous esophageal neoplasm in patients with hypopharyngeal squamous cell carcinoma (HSCC). Methods: A retrospective review of 96 cases with HSCC that received surgical treatment from March 2016 to March 2017 was accomplished. In these patients, the site of origin were pyriform sinus (n=75), posterior pharyngeal wall (n=11) and postcricoid (n=10). Esophagoscopy was prospectively performed on all patients before treatment for HSCC. All patients underwent conventional white-light endoscopic examination with Lugol chromoendoscopy and narrow band image. Suspicious areas of narrow band image or Lugol-voiding lesions were observed and biopsied. The treatment strategy of primary HSCC was modified according to the presence of synchronous esophageal squamous cell neoplasms by a multidisciplinary approach. Results: Ninety-six patients were enrolled (age ranging from 37-80 years). All patients did not have previous treatment.Histopathological analysis revealed middle to high-grade dysplasia in 5 cases, Tis cancer in 5 cases, cancer in 16 cases and inflammation or normal findings in the others. Four cases were treated with endoscopic submucosal dissection before hypopharygeal surgery, 3 cases with lower esophageal cancers were treated with gastric pull-up combined with free jejunal flap after total circumferential pharyngolaryngectomy (TCPL) and certical esophagectomy, and 14 cases were treated with TCPL, total esophagectomy and gastric pull-up. Conclusions: Esophagoscopy is a feasible and justified procedure in HSCC cases as it enhances the detection of premalignant lesion or second primary cancer. Routine esophagoscopy for detecting synchronous second primary tumor should be recommended for patients with HSCC. The treatment strategy for primary HSCC is modified according to the presence of synchronous second primary tumor.

Association of DGAT2 gene polymorphisms with carcass and meat quality traits in domestic pigeons (Columba livia).

1. Diacylglycerol acyltransferase (DGAT) plays an important role in the synthesis of triacylglycerol, but its effects on meat quality and carcass composition in pigeons are unclear. In this study, single-nucleotide polymorphisms (SNPs) in the exons of the DGAT2 gene were identified and analysed by using DNA sequencing methods in 200 domestic pigeons (Columba livia). The associations between DGAT2 polymorphisms and carcass and meat quality traits were also analysed. 2. Sequencing results showed that 5 nucleotide mutations were detected in exons 3, 4, 5 and 6 of the DGAT2 gene. The analysis revealed three genotypes (AA, AB and BB) in G18398T and G22484C, in which the AA genotype and A allele had the highest frequency. 3. In the SNP of G18398T located in exon 5, individuals with genotype BB had significantly higher meat quality and lower abdominal fat content than those with AA or AB genotype. In the SNP of G22484C located in exon 6, the genotype AA showed highest carcass trait values, while the genotype BB represented better meat quality, compared to AA and AB genotypes. 4. The results imply that DGAT2 gene has a close relationship with carcass and meat quality traits in pigeons, and the SNPs of G18398T and G22484C can be used as genetic markers for marker-assisted breeding in pigeon.

Effects of rearing systems on laying performance, egg quality, and serum biochemistry of Xianju chickens in summer.

The objective of this study was to compare the laying performance, egg quality, and serum biochemistry of hens maintained in conventional cage rearing system (CRS), flat net-rearing system (NRS), and free range system (FRS) under summer conditions. Indigenous Xianju chickens (n = 540) were randomly allocated into cages or pens of rearing system groups, within each system there were 5 replicates with 36 hens in each replicate. The experiment lasted between 21 and 29 wk of age. Hen-day egg production (P = 0.00) and egg mass (P = 0.00) were higher in the CRS but were similar in the NRS and FRS. Lowest egg weight (P = 0.02), yolk weight (P = 0.00) and yolk ratio (P = 0.01), and feed intake (P = 0.01) were observed from the FRS, whereas lowest feed conversion ratio (FCR) was recorded from the CRS (P = 0.01). Rearing systems had negligible effect on egg quality. Serum Ca (P = 0.04) and total protein (P = 0.03) levels were found to be higher in the CRS but were lower in the FRS. Serum levels of glucose (P = 0.01), cholesterol (P = 0.00), and triglyceride (P = 0.00) in the CRS increased compared with the NRS and FRS groups, whereas serum levels of high density lipoprotein cholesterol (HDL-C; P = 0.01) in the CRS decreased. It can be concluded that under summer conditions, Xianju chickens from CRS had an advantage in terms of productivity parameters, but exhibited higher levels of serum lipids and glucose.

miR-146a rs2910164 polymorphism might be associated with coronary artery disease risk in Asians.

Recently, several studies reported the association between miR-146a rs2910164 polymorphism and coronary artery disease (CAD) risk. However, the results were inconclusive. We therefore did a meta-analysis to investigate this association. Electronic databases, including PubMed, EMBASE, and China National Knowledge Infrastructure (CNKI) databases, were searched. The strength of association was assessed by calculating odds ratios (OR) and 95% confidence interval (CI). Five eligible studies included in this meta-analysis. The total sample size of this meta-analysis was 1565 cases and 1541 controls. We found that miR-146a rs2910164 polymorphism was significantly associated with an increased CAD risk (OR = 1.19; 95% CI 1.07 - 1.32; P = 0.002). In the subgroup analysis by race, miR-146a rs2910164 polymorphism was significantly associated with an increased CAD risk in Asians (OR = 1.18; 95% CI 1.04 - 1.33; P = 0.008). However, we did not find significant result in Caucasians (OR = 1.22; 95% CI 0.86 - 1.74; P = 0.25). In the subgroup analysis by age, we found that miR-146a rs2910164 polymorphism increased CAD risk in old population (OR = 1.22; 95% CI 1.09 - 1.38; P = 0.0008). In conclusion, this meta-analysis found that miR-146a rs2910164 polymorphism was significantly associated with CAD risk.

Is differential expression of p16INK4a based on the classification of uterine smooth muscle tumors associated with a different prognosis? A meta-analysis.

We conducted a meta-analysis to examine p16INK4a expression in uterine smooth muscle tumors (USMTs). Although the prognostic value of tumor suppressor p16INK4a has been elucidated in a variety of cancers and precancerous lesions, its role in USMTs is not well established. We searched PubMed, Web of Science, and Embase for publication son p16INK4a expression in USMTs. Strict inclusion and exclusion criteria were imposed. Risk ratios (RRs) with 95% confidence intervals (95%CIs) were calculated to assess the strength of association. Publication bias was estimated using funnel plots and the Egger's regression test. Twelve eligible studies comprising 661 patients were included. Compared with leiomyoma (LM), the figures for the strength of association were as follows: LM variants (RR = 1.53, 95%CI = 1.03-2.27, P = 0.036, random effect); leiomyosarcoma (LMS) (RR = 3.20, 95%CI = 1.68-6.12, P < 0.001, random effect); and smooth muscle tumors of uncertain malignant potential (STUMP) (RR = 2.90, 95%CI = 1.17-7.21, P = 0.022, random effect). p16INK4a expression was significantly higher in LMS than in LM variants (RR = 3.74, 95%CI = 1.96-7.13, P < 0.001, random effect) or STUMP (RR = 1.67, 95%CI = 1.26-2.23, P < 0.001, fixed effect). There was a significant correlation between overexpressed p16INK4a and recurrence rates of USMTs (RR = 1.85, 95%CI = 1.11-3.10, P = 0.019, fixed effect). p16INK4a over expression is a potential biomarker for diagnosing problematic USMTs and it might indicate a worse prognosis. However, there is currently insufficient evidence to assess the prognostic value of p16INK4a in USMTs.

In Vitro and In Vivo Antibacterial Activities of Patchouli Alcohol, a Naturally Occurring Tricyclic Sesquiterpene, against Helicobacter pylori Infection.

This study further evaluated the in vitro and in vivo anti-Helicobacter pylori activities and potential underlying mechanism of patchouli alcohol (PA), a tricyclic sesquiterpene. In the in vitro assay, the capacities of PA to inhibit and kill H. pylori were tested on three standard strains at different pH values and on 12 clinical isolates. The effects of PA on H. pylori adhesion (and its alpA, alpB, and babA genes), motility (and its flaA and flaB genes), ultrastructure, and flagellation were investigated. Moreover, the H. pylori resistance to and postantibiotic effect (PAE) of PA were determined. Furthermore, the in vivo effects of PA on H. pylori eradication and gastritis were examined. Results showed that MICs of PA against three standard strains (pH 5.3 to 9) and 12 clinical isolates were 25 to 75 and 12.5 to 50 µg/ml, respectively. The killing kinetics of PA were time and concentration dependent, and its minimal bactericidal concentrations (MBCs) were 25 to 75 µg/ml. In addition, H. pylori adhesion, motility, ultrastructure, and flagellation were significantly suppressed. PA also remarkably inhibited the expression of adhesion genes (alpA and alpB) and motility genes (flaA and flaB). Furthermore, PA treatment caused a longer PAE and less bacterial resistance than clarithromycin and metronidazole. The in vivo study showed that PA can effectively eradicate H. pylori, inhibit gastritis, and suppress the expression of inflammatory mediators (COX-2, interleukin 1ß, tumor necrosis factor alpha, and inducible nitric oxide synthase [iNOS]). In conclusion, PA can efficiently kill H. pylori, interfere with its infection process, and attenuate gastritis with less bacterial resistance, making it a potential candidate for new drug development.

[Effect of exendin-4 on lipid deposition in skeletal muscle of diet-induced obese mice and its underlying mechanism].

Objective: To investigate the effect of exendin-4, a glucagon-like peptide-1 (GLP-1) receptor agonist, on reducing lipid deposition and improving insulin resistance in skeletal muscle and the underlying mechanisms in high-fat diet (HFD)-induced obese mice. Methods: Twelve male C57BL/6J mice were challenged with HFD for 12 weeks to induce obesity and then randomly divided into two groups: exendin-4 group (intraperitoneal injection of 24 nmol·kg-1·d-1 exendin-4 for 4 weeks) and HFD group (intraperitoneal injection of normal saline for 4 weeks), with 6 mice in each group. Additional 6 mice were also selected as control group. Body weight, fasting blood glucose were recorded. Serum triglyceride (TG), total cholesterol (TC), insulin and skeletal muscle triglyceride levels were measured by enzyme-linked immunosobent assay (ELISA). Oil red O staining was used for morphologic changes of frozen sections from skeletal muscle. The protein levels of lipid metabolic pathway mediated by AMP-activated protein kinase (AMPK) and insulin signailing pathway were determined by Western blot. Results: Compared with mice in HFD group, exendin-4 significantly decreased body weight[(37.68±1.80) vs (46.03±5.00) g, P<0.025], fasting blood glucose[(5.40±0.33) vs (7.65±1.92) mmol/L, P<0.025], serum TG[(37.78±7.14) vs (80.76±34.22) mg/dl, P<0.025], TC[(180.13±18.75) vs (217.57±22.52) mg/dl, P<0.025], insulin[(0.58±0.01) vs (1.67±1.23) ng/ml, P<0.025]and skeletal muscle TG levels[(9.84±1.08) vs (19.35±7.44) mg/g, P<0.025]of obese mice. Oil red O staining revealed that exendin-4 alleviated the accumulation of larger lipid droplets in skeletal muscle. The protein expressions of lipolysis and lipid oxidation mediated by AMPK and insulin signailing pathway were up-regulated, and the protein expressions of lipogenesis mediated by AMPK were down-regulated after intervention of exendin-4. Conclusion: Exendin-4 reduces lipid deposition and insulin resistance in skeletal muscle of HFD-induced obese mice via activating AMPK and up-regulating insulin signailing pathway.