Ammonium depletion associated with the COVID-19 pandemic in the Mexican Caribbean.

The Mexican Caribbean contributes significantly to Mexico's gross national product. The number of tourists declined from 16.7 million in 2019 to 8.8 million in 2020 due to the COVID-19 pandemic, with a rapid recovery of 13.5 million in 2021. Wastewater discharge is the primary contamination source associated with the tourism sector's demand for goods and services. Water quality could improve due to fewer tourists arriving during the COVID-19 sanitary emergency. This study aimed to quantify ammonium concentrations at eleven locations to evaluate water quality during the sanitary restriction due to the pandemic in the Mexican Caribbean. The ammonium concentrations were 85 % (Nov-2019), 89 % (Feb-2020), and 86 % (Feb-2021) higher than in Nov-2020, where six of the eleven sampled stations were below the detection limit (0.15 µM). Lower ammonium concentrations coincide with the sanitary restriction period and a decrease in affluent tourists.

Identification of the function of γδ1 T cells in the lung cancer microenvironments.

PURPOSE: To investigate whether γδ1 T cells derived from lung cancer tissues have immunosuppressive function and to verify the mechanism of immunosuppressive effect. METHODS: Fresh lung cancer tissue samples were collected, some of them were prepared tissue sections, the others were isolated and amplified into TILs cells, γδ1 T cells were isolated from TILs cells by immunomagnetic beads kits, and then cloned and amplified. The immunomodulatory effects of γδ1 T cells on naive and effector CD4+ T cells were detected by immunohistochemistry, flow cytometry, CCK8, ELISA and transwell culture. RESULTS: A high proportion of γδ1 T cells was found in lung cancer tissues. The cultural supernatants of γδ1 T cells could inhibit the proliferation of naive CD4+ T cells and decrease the secretion level of IL-2 by effector CD4+ T cells. Further studies showed that the expression levels of IL-8, MIP-1α, MIP-1ß and RANTES were higher than that of IFN-γ, GM-CSF and TNF-α, TNF-ß, however, their neutralizing antibodies could not block the immunosuppressive activity of the supernatant. CONCLUSION: γδ1 T cells play an negative immunoregulation function in lung cancer microenvironments, and have obvious immunosuppressive effects on proliferation and cytokine release of naive CD4+ T cells and effector CD4+ T cells. Preliminary evidence from this study suggests that the mechanism of immunosuppressive effects is mediated by the soluble factors in γδ1 T cell culture supernatants, but its exact molecular mechanism needs to be further explored.

circRNA CRIM1 regulates the migration and invasion of bladder cancer by targeting miR182/Foxo3a axis.

PURPOSE: To explore the molecular mechanism of circRNA CRIM1 in the regulation of bladder cancer by targeting the miR182/Foxo3a axis. METHODS: 50 pairs of cancer tissues and para-cancerous tissues of patients with bladder cancer were collected. RT-PCR method was used to detect the expression of CRIM1 and miR-182. The association between circRNA CRIM1 and clinical data was analyzed. qPCR was used to measure the expression of circRNA CRIM1 and miR-182 in bladder cancer cell UMUC3 and endothelial cell line HUVEC. CRIM1 genes and miR-182 in UMUC3 cell lines were overexpressed and silenced, respectively, to investigate their effects on invasion and migration of bladder cancer, and to detect the changes of miR182/Foxo3a expression. The association between circRNA CRIM1 and miR182/Foxo3a was determined by bioinformatics analysis. RESULTS: The results showed that there was a significant association between the expression of circRNA CRIM1 and distal migration. The expression of CRIM1 in adjacent tissues was significantly down-regulated and negatively correlated with distal migration. The overexpression of circRNA CRIM1 reduced migration and invasion processes in bladder cancer cells. After circRNA CRIM1 was overexpressed, the miR-182 was significantly down-regulated. The expression levels of Foxo3a mRNA and proteins were up-regulated after miR-182 silencing of bladder cancer cell line UMUC3. miR-182 silencing inhibited invasion and migration of cancer cells to some extent. In bladder cancer cells and tissues, CRIM1 and Foxo3a were significantly down-regulated, miR-182 was significantly up-regulated. CONCLUSION: circRNA CRIM1 regulated the migration and invasion of bladder cancer by targeting the miR182/Foxo3a axis.

UCHL3 promotes aerobic glycolysis of pancreatic cancer through upregulating LDHA expression.

BACKGROUND: Aerobic glycolysis has a pivotal role in the carcinogenic process. The current understanding of the functional role and mechanism of UCHL3-related aerobic glycolysis in pancreatic cancer is far from comprehensive, therefore requires an in-depth analysis on this aspect. METHODS: In the present research, the expressions of ubiquitin carboxyl-terminal hydrolase L3 (UCHL3), lactate dehydrogenase A (LDHA) and Forkhead box protein M1 (FOXM1) were detected by qRT-PCR, Western blot and immunohistochemistry. The effects of UCHL3 knockdown or overexpression on pancreatic cancer cells were examined by determining cell viability and colony formation. Aerobic glycolysis was assessed according to glucose uptake, lactic acid production, and lactate dehydrogenase (LDH) activity. Dual-luciferase reporter assay was performed to detect LDHA promoter activity. RESULTS: The results showed that UCHL3 expression was significantly increased in the pancreatic cancer tissues and cells, and that knocking down UCHL3 noticeably inhibited cell viability and aerobic glycolysis. Further investigations revealed that LDHA expression was promoted by UCHL3 and could be reduced by shFOXM1, and that low-expressed LDHA partly reversed the inhibition of aerobic glycolysis induced by overexpressed UCHL3. CONCLUSIONS: To conclude, this study demonstrates that UCHL3 plays a carcinogenic role by promoting aerobic glycolysis in pancreatic cancer, suggesting that UCHL3 may be a potential diagnostic and therapeutic target for the treatment of cancer.

Isolation and characterization of polymorphic microsatellite markers for the chub mackerel (Scomber japonicus) and cross-species amplification in the blue mackerel (S. australasicus).

In this study, 10 polymorphic microsatellite markers were developed in Scomber japonicus and were examined on 30 individuals collected from the North Pacific. The number of alleles per locus ranged from 4 to 17. The observed and expected heterozygosities per locus ranged from 0.2759 to 0.8621 and from 0.43071 to 0.9177, respectively. The polymorphism information content (PIC) was from 0.3931 to 0.8939. One locus showed moderate polymorphism (0.25 < PIC < 0.5), while the rest were highly polymorphic (PIC > 0.5). Two loci showed significant deviation from Hardy-Weinberg equilibrium after Bonferroni corrections (P < 0.005). No linkage disequilibrium was detected among the loci. Results of cross-species amplification showed that 10 microsatellite markers were successfully amplified in 29 individuals of S. australasicus and 9 indicated polymorphisms. These markers will be useful for investigating the genetic structure, gene flow, and species identification of S. japonicus and S. australasicus, its closely related species.

MicroRNAs Association in the Cardiac Hypertrophy Secondary to Complex Congenital Heart Disease in Children.

Complex congenital heart disease (CHD) affects cardiac blood flow, generating a pressure overload in the compromised ventricles and provoking hypertrophy that over time will induce myocardial dysfunction and cause a potential risk of imminent death. Therefore, the early diagnosis of complex CHD is paramount during the first year of life, with surgical treatment of patients favoring survival. In the present study, we analyzed cardiac tissue and plasma of children with cardiac hypertrophy (CH) secondary to CHD for the expression of 11 miRNAs specific to CH in adults. The results were compared with the miRNA expression patterns in tissue and blood of healthy children. In this way, we determined that miRNAs 1, 18b, 21, 23b, 133a, 195, and 208b constitute the expression profile of the cardiac tissue of children with CHD. Meanwhile, miRNAs 21, 23a, 23b, and 24 can be considered specific biomarkers for the diagnosis of CH in infants with CHD. These results suggest that CH secondary to CHD in children differs in its mechanism from that described for adult hypertrophy, offering a new perspective to study the development of this pathology and to determine the potential of hypertrophic miRNAs to be biomarkers for early CH.

Association of neutrophil/lymphocyte ratio and platelet/lymphocyte ratio with ER and PR in breast cancer patients and their changes after neoadjuvant chemotherapy.

PURPOSE: Proinflammatory markers, including neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR), are associated with many aspects of different malignancies. The aim of this study was to assess the associations of NLR and PLR with estrogen receptor (ER) and progesterone receptor (PR) expression in locally advanced breast cancer patients and their changes after neoadjuvant chemotherapy (NAC). Whether these parameters were predictive for the response to NAC in breast cancer patients was also evaluated. METHODS: 132 Female primary locally advanced breast cancer patients treated with either ET (epirubicin-docetaxel), TEC (docetaxel-epirubicin-cyclophosphamide), or CEF (cyclophosphamide-epirubicin-fluorouracil) as NAC were retrospectively studied. NLR and PLR were calculated from peripheral blood cell count and their optimal cutoff levels were determined by receiver operating characteristic curves. RESULTS: The proportion of ER-positive breast cancers before NAC was higher both in NLRlow (<2.05) group and PLRlow group (<159.01). Changes in ER or PR expression level or status were observed in some patients. The alterations of NLR and PLR after NAC correlated with chemotherapy regimens, and elevated PLR was found. The patients with low pretreatment NLR (<1.67) or PLR (<151.27) had better responses to NAC than those with high NLR (≥1.67, 67.3 vs. 47.1%, P < 0.05) or PLR (≥151.27, 64.0 vs. 45.1%, P < 0.05). CONCLUSIONS: The patients with low pretreatment NLR (<2.05) or PLR (<159.01) had higher ER expression. Changes in ER and PR expression status or level occured following NAC. Elevated PLR was found aft-NAC. Pretreatment NLR and PLR may be important predictive indicators for NAC response in breast cancer patients.

Meristic and morphometric analysis of two hagfish species (Myxine affinis and Notomyxine tridentiger) from the Magellan Strait, Chile / Análisis nerístico y morfométrico de dos especies de anguila babosa (Myxine affinis y Notomyxine tridentiger) en el Estrecho de Magallanes, Chile

Myxinoids in Chile are represented by the subfamilies Eptatretinae and Myxininae, with a total of 14 species, the identification is complex due to the low level of morphological differentiation that characterizes this taxonomic group. Worldwide, hagfish are species of commercial value, and in Chile many attempts have been reported to initiate small-scale fisheries. The aim of the present study is describe the hagfish species caught in an incipient fishery of the Magellan Strait. Samples were collected in the Magellan Strait during eight fishing expeditions from June 2009 to October 2010 in Bahía Lomas (5348`S; 70°46'W) and Agua Fresca (5323`S; 70°45'W). The samples were taken at two depths, 0-70 meters and 71-140 meters. Taxonomic keys were used to identify the species. All specimens were individuals from the Myxininae subfamily. From a total of 3946 hagfishes, 99 % (n=3905) were the species Myxine affinis and the remaining 1 % were Notomyxine tridentiger, both reported for Chilean and Argentinean Patagonia. The range of variation for meristic variables recorded in this research was wider than those reported in the literature. This could be explained by differences in sample size between the present study and those previously published. Body proportions and meristic variables were not species specific in Myxine sp, so there is a large overlap of ranges between species, which makes their diagnostic use not applicable. This research updates the information and extends the meristic ranges for both species. The esophageocutaneous duct (in N. tridentiger) and the number of fused teeth (bicuspid in M. affinis and tricuspid in N. tridentiger) are the morphological characters that allow a clear identification in the field of the two species.

Los Myxinoideos en Chile están representados por las subfamilias Eptatretinae y Myxininae, con un total de 14 especies, cuya identificación resulta compleja debido al bajo nivel de diferenciación morfológica que caracteriza a este grupo taxonómico. A nivel mundial las anguilas babosas constituyen especies de valor comercial, y en Chile se reportan varios intentos para iniciar pesquerías de pequeña escala. El presente estudio tuvo como propósito la descripción de las especies de anguila babosa capturadas en una pesquería incipiente del Estrecho de Magallanes. Los ejemplares fueron colectados durante ocho expediciones de pesca, desde junio de 2009 a octubre de 2010 en Bahía Lomas (5348`S; 70°46'W) y Agua Fresca (5323`S; 70°45'W). Las muestras fueron tomadas en dos rangos de profundidad, 0-70 metros y 71-140 metros. Todos los especímenes fueron pertenecientes a la subfamilia Myxininae. De un total de 3946 anguilas el 99 % (n=3905) pertenecieron a la especie Myxine affinis y el restante 1 % a la especie Notomyxine tridentiger, ambas reportadas para Chile y la Patagonia Argentina. El rango de variación para las variables merísticas, registradas en esta investigación, fue mayor a los reportados en literatura. Esto puede ser explicado por las diferencias en el tamaño de muestra entre el presente estudio y aquellos publicados previamente. Las proporciones corporales y las variables merísticas no fueron especie-específica en Myxine sp, por lo que existe una gran sobre posición de los rangos entre las especies, lo que hace que su uso como diagnóstico no sea aplicable. Esta investigación actualiza la información y extiende los rangos merísticos para ambas especies. El conducto esofágico-cutáneo (en N. tridentiger) y el número de dientes fusionados (bicúspide en M. affinis y tricúspide en N. tridentiger), son los caracteres morfológicos que permiten una identificación clara de ambas especies.

Prognostic significance of immune cells in the tumor microenvironment and peripheral blood of gallbladder carcinoma patients.

BACKGROUND: The role of the interaction between tumor cells and inflammatory cells in gallbladder carcinoma (GBC) is unclear. Inflammatory cells exist in both the tumor immune microenvironment and the host peripheral blood circulatory system. In the current study, we examined the prognostic value of inflammatory cells in the tumor microenvironment and peripheral blood in patients with GBC. METHODS: 98 patients with GBC were recruited in this retrospective study. Using immunohistochemistry, we examined tumor-infiltrating CD3+ generic T-cells, CD8+ cytotoxic T-cells, CD45RO+ memory T-cells, and CD15+ neutrophils. Peripheral venous blood samples were also collected, and absolute neutrophil count (ANC), absolute lymphocyte count (ALC) and neutrophil/lymphocyte ratio (NLR) were measured. The relationships between these variables and patient outcome were evaluated. RESULTS: Survival analysis revealed that the density of CD3+ cell infiltrates in the tumor microenvironment was positively correlated with overall survival (OS) and the density of CD15+ cell infiltrates was negatively correlated with the OS. The combined analysis showed that a high density of CD3+ cell infiltrates combined with a low density of CD15+ cell infiltrates was an independent prognostic factor for GBC. In peripheral blood, survival analysis suggested that ANC and NLR were negatively correlated, while ALC was positively correlated with OS. Multivariate survival analysis showed that NLR was an independent prognostic factor for gallbladder cancer prognosis. CONCLUSIONS: The results indicate that the combination of high density of CD3+ cell infiltrates combined with a low density of CD15+ cell infiltrates in tumor samples and pretreatment peripheral blood NLR were independent prognostic factors in patients with GBC.

Characterization of the complete mitochondrial genome of Portunus pelagicus with implications for phylogenomics.

This study determined the mitochondrial genome structure of the blue swimming crab (Portunus pelagicus), and elucidated its phylogenetic relationships among the species within the order Decapoda. The complete mitochondrial genome was 16,155 bp long, and contained 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 DNA control region. The gene order of the genome was the same as that found within the family Portunidae. Twenty-three genes were on the heavy strand and 14 were on the light strand. Almost all of the protein-coding genes were initiated by an ATG codon, except for three genes (ATP6, ND1, and ND3) that started with a rare ATT codon. Of the 13 protein-coding genes, 10 ended with complete TAA or TAG stop codons and three ended with an incomplete T codon. Thirteen non-coding regions were identified that ranged from 1 to 30 bp in length. Nine overlaps were found, which ranged 1 to 7 bp in length. Phylogenetic analyses based on 12 concatenated protein-coding genes revealed that P. pelagicus formed a monophyletic group with Portunus trituberculatus, which were in a larger group with Callinectes sapidus, while the genera Charybdis and Thalamita formed another group. These two groups clustered together and grouped with the genus Scylla. The phylogenetic analysis supported the inclusion of Charybdis in subfamily Portuninae of the family Portunidae, and revealed a close relationship between Charybdis and Thalamita. We suggest that Thalamita should also be classified into the subfamily Portuninae. The results can be used in the study of phylogenetic, population genetic and conservation genetics of P. pelagicus.

Optimization of selective breeding through analysis of morphological traits in Chinese sea bass (Lateolabrax maculatus).

Determining correlations between certain traits of economic importance constitutes an essential component of selective activities. In this study, our aim was to provide effective indicators for breeding programs of Lateolabrax maculatus, an important aquaculture species in China. We analyzed correlations between 20 morphometric traits and body weight, using correlation and path analyses. The results indicated that the correlations among all 21 traits were highly significant, with the highest correlation coefficient identified between total length and body weight. The path analysis indicated that total length (X1), body width (X5), distance from first dorsal fin origin to anal fin origin (X10), snout length (X16), eye diameter (X17), eye cross (X18), and slanting distance from snout tip to first dorsal fin origin (X19) significantly affected body weight (Y) directly. The following multiple-regression equation was obtained using stepwise multiple-regression analysis: Y = -472.108 + 1.065X1 + 7.728X5 + 1.973X10 - 7.024X16 - 4.400X17 - 3.338X18 + 2.138X19, with an adjusted multiple-correlation coefficient of 0.947. Body width had the largest determinant coefficient, as well as the highest positive direct correlation with body weight. At the same time, high indirect effects with six other morphometric traits on L. maculatus body weight, through body width, were identified. Hence, body width could be a key factor that efficiently indicates significant effects on body weight in L. maculatus.

Characterization of the complete mitochondrial genome and phylogenetic relationships of the three-spot swimming crab (Portunus sanguinolentus).

In this study, we determined the whole mitochondrial genome profile of the three-spot swimming crab (Portunus sanguinolentus) and elucidated phylogenetic relationships between representative species in the order Decapoda. The mitochondrial genome was 16,024 bp in length and consisted of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a putative control region. Of the 37 genes, 23 were encoded by the heavy strand while 14 were encoded by the light strand. Four types of start codons were identified; ATG initiated nine genes, ATT initiated two genes, and ATC and GTG each started one gene. Nine protein-coding genes ended with a complete TAA or TAG stop codon, and four genes ended with an incomplete T or TA codon. Fourteen non-coding regions were found, which ranged from 1 to 34 bp in length. Nine overlaps were observed, with lengths between 1 and 7 bp. Phylogenetic analysis suggested that P. sanguinolentus is genetically closest to P. trituberculatus and P. pelagicus. Charybdis feriata, C. japonica, and Thalamita crenata formed a single cluster, and were close to the genera Callinectes and Portunus. Therefore, the genera Charybdis and Thalamita should be classified into the subfamily Portuninae.

First genetic linkage map for the mud crab (Scylla paramamosain) constructed using microsatellite and AFLP markers.

The mud crab (Scylla paramamosain) is of economic importance for the fisheries and aquaculture industry in China. In this study, we constructed the first genetic linkage map for this species using microsatellite and amplified fragment length polymorphism (AFLP) markers. The map consisted of 65 linkage groups, including 34 triplets and 9 doublets. A total of 212 molecular markers were mapped, including 60 microsatellites and 152 AFLP markers. The linkage groups ranged from 7 to 102.5 cM and covered 2746.4 cM in length. The mean length was 42.3 cM per linkage group, and the mean spacing was 18.68 cM. The genome size was estimated to be 5539.62 cM, with 50% coverage by the present map. Moreover, we reported 5 transcriptome-derived polymorphic microsatellite markers and characterized their polymorphism in a first-generation family. This study will facilitate studies on high-density maps and molecular marker-assisted selection in S. paramamosain and related crustacean species.

[Lymphocele surgical treatment in pediatric kidney transplantation]. / Tratamiento quirúrgico de linfocele en trasplante renal pediátrico.

BACKGROUND: In a retrospective study, we evaluated; frequency, clinical presentation and treatment of lymphocele in pediatric patients with kidney transplant. METHODS: Between January 2004 and January 2009, we had 242 kidney transplantations, 197 from living donors and 45 from cadaveric donors. The technique was the usual, and the implants of the ureteres were by the technique Ricard modified. The treatment of lymphocele was by percutaneous punction and laparoscopic intraperitoneal drainage. RESULTS: We diagnosed lymphocele in seven patients (2.9 % with an IC95 %: 0.6- 5.2 %) one female and six males. All patients went trough percutaneous drainage. Six patients presented lymphocele recurrence at 48 to 72 hours after the drainage. Surgical laparoscopic intraperitoneal drainage was performed with success. CONCLUSIONS: Our frequency of lymphocele is the same reported in other studies (6-18 %). The treatment by laparoscopic intraperitoneal window seems to be the most appropriate in pediatric patients.

Introducción: en un estudio observacional, retrospectivo evaluamos la frecuencia, presentación clínica y tratamiento de linfocele en pacientes pediátricos con trasplante renal. Métodos: de enero de 2004 a enero de 2009 se realizaron 242 trasplantes renales, 197 de donantes vivos y 45 trasplantes de donantes cadavéricos. La técnica quirúrgica utilizada fue la habitual y la de los implantes uretrales fue la técnica Ricard modificada. El tratamiento fue por punción cutánea y drenajes internos mediante una ventana peritoneal laparoscópica. Resultados: se diagnosticaron siete pacientes con linfoceles (2.9 % con un IC 95 % 0.6-5.2 %). Una paciente del sexo femenino y seis del sexo masculino, todos fueron sometidos a drenajes por punción cutánea, seis pacientes fueron recidivados y tratados exitosamente por drenaje interno mediante una ventana peritoneal laparoscópica. Conclusiones: nuestra frecuencia es igual a la reportada en otros estudios (0.6-18 %). El drenaje interno mediante la ventana peritoneal laparoscópica parece ser lo más apropiado en pacientes pediátricos.

Pre-harvest foliar application of ethephon strengthens gibberellins-induced fruit expansion in Pyrus pyrifolia.

To identify the roles of ethylene in fruit development in Japanese pear Pyrus pyrifolia 'Niitaka', one of the non-climacteric genotypes, source-sink strength and fruit development during fruit expansion were investigated when ethephon was applied after a conventional gibberellic acid (GA) lanolin paste treatment on the pedicel. The results demonstrate that the conventional GA treatment during the early stage of fruit expansion resulted in larger fruit size and advanced fruit maturation, but pre-harvest foliar application of ethephon only advanced fruit maturation. However, pre-harvest foliar application of ethephon with a preceding conventional GA treatment during the early stage of fruit expansion dramatically improved fruit size and advanced fruit maturation over GA or ethephon alone. Moreover, the early foliar application of ethephon showed a better efficacy in increasing fruit size than the late spraying. A further study revealed that when ethephon was applied after the conventional GA treatment, it improved source-sink strength associated with leaf photosynthesis and the specific rate of [13C] accumulation in fruit, and also strengthened cell expansion more than did GA or ethephon alone.

Discovery and characterization of a first set of polymorphic microsatellite markers in Siganus oramin.

Nine microsatellite DNA markers were developed and characterized for Siganus oramin by the 5'-anchored polymerase chain reaction technique. A total of 42 alleles were identified in 30 individuals, and the number of alleles per locus ranged from 3 to 7, with an average of 4.7. The observed and expected heterozygosity per locus ranged from 0.5333 to 1.0000 and from 0.5254 to 0.8474, respectively, with an average of 0.7422 and 0.6906, respectively. A significant deviation from the Hardy-Weinberg equilibrium was detected at one microsatellite locus after a Bonferroni's correction (P < 0.0056). No significant linkage disequilibrium was found between any of the pairs of the nine loci. The microsatellite loci developed in this study will improve our understanding of the genetic background of S. oramin.

Role of VEGF gene polymorphisms in the clinical outcome of non-small cell lung cancer.

This report details a study conducted to assess the role of VEGF gene polymorphisms in the prognosis of advanced non-small cell lung carcinoma (NSCLC). Samples obtained from 210 advanced NSCLC patients admitted at the Huaihe Hospital of Henan University between January 2010 and December 2011 were recruited for this study. The VEGF -2578C/A (rs699947), +936C/T (rs3025039), and -634G/C (rs2010963) genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism. We discovered, by logistic regression analysis, that the TT genotype of VEGF +936C/T was associated with more complete response + partial response to chemotherapy, compared to the CC genotype (odds ratio (OR) = 4.78, 95% confidence interval (CI) = 1.34-25.85). We also found a correlation between the TT genotype of VEGF +936C/T and lower risk of death from all causes compared to the CC genotype (OR = 0.26, 95%CI = 0.10-0.69), using the Cox proportional hazard model (after adjusting for potential confounding factors). In conclusion, we discovered that the VEGF +936C/T gene polymorphism influences the response to chemotherapy and overall survival of NSCLC patients.

Molecular authentication of Gynostemma pentaphyllum through development and application of random amplification polymorphic DNA sequence-characterized amplified region marker.

Due to the morphological similarities of aerial parts, it is difficult to distinguish Gynostemma pentaphyllum from Cayratia japonica, which is usually an adulterant of the former. To develop a reliable method for the identification and authentication of G. pentaphyllum, a combination of random amplification polymorphic DNA (RAPD) technique with sequence-characterized amplified region (SCAR) markers was studied. Twenty-five samples of G. pentaphyllum and two samples of C. japonica were collected from different regions in Guangxi or bought from different provinces in China. Through the RAPD analysis, significant genetic polymorphism was observed among the intraspecies samples of G. pentaphyllum. Furthermore, a specific marker, J-750, was obtained for authentication. Therefore, the SCAR marker for G. pentaphyllum (359 bp) was developed from the RAPD amplicon. With PCR amplification using the SCAR primers, a specific band of 359 bp was distinctly visible for all tested samples of G. pentaphyllum, but was absent in the samples of C. japonica. Furthermore, the results revealed that the SCAR marker was useful for the identification and authentication of G. pentaphyllum irrespective of whether samples were fresh, dry, or of commercial origin. The SCAR marker obtained in this study successfully authenticated G. pentaphyllum through an integrated PCR system containing SCAR and control primer combinations of two pairs. In addition, it was also used for simultaneous discrimination of G. pentaphyllum from C. japonica.

Risk Factors for Death of Elderly Patients with Acute Obstructive Suppurative Cholangitis.

OBJECTIVE: To identify the risk factors for death of elderly patients with acute obstructive suppurative cholangitis (AOSC). METHODS: Three hundred and forty-eight AOSC patients > 60 years of age were retrospectively analysed in the First People's Hospital of Jining from June 2005 to June 2013. The patients were treated with endoscopic retrograde cholangiopancreatography (ERCP) immediately after AOSC was diagnosed to clear the stones and drain, and surgical procedures were then performed in the patients in whom ERCP failed. The risk factors were identified with univariate and multivariate analysis. RESULTS: Among the 348 AOSC patients, 27 patients died after treatment. Two hundred and forty-nine patients were treated with ERCP, and 11 patients died; 99 patients were treated with ERCP plus surgery, and 16 patients died. Two hundred and thirty-two patients were treated within 24 hours after they were admitted to the hospital, and 10 patients died; 116 patients were treated beyond 24 hours, and 17 patients died. According to the results of the univariate and multivariate analysis, shock, ERCP plus surgery, advanced age, low platelet count, the presence of co-morbidities, door to treatment time > 24 hours, hypoproteinaemia, and hyperbilirubinaemia were the independent risk factors for death of elderly patients with AOSC. CONCLUSION: The strategies of dealing with these risk factors should be researched to reduce mortality of elderly patients with AOSC.

Mutation analysis of four Chinese families with pure hereditary spastic paraplegia: pseudo- X-linked dominant inheritance and male lethality due to a novel ATL1 mutation.

We studied four Chinese families with pure hereditary spastic paraplegia (HSP) to investigate the clinical features and associated genetic mutations. Linkage analysis was performed for all families to map the disease locus onto autosomal chromosomes, and related loci involved in HSP on the X chromosome were also examined. Polymerase chain reaction (PCR) sequencing was used to detect gene mutations. To confirm the influence of a splice-site mutation on mRNA, we used reverse transcription-PCR and direct sequencing. Linkage analysis and ATL1 gene sequencing of amniocytes were performed for prenatal genetic diagnosis. One missense variant (c.1517T>A) and a splice-site mutation (c.1245+1G>A) in SPAST, and two missense variants (c.715C>T, c.1204T>G) in ATL1 were identified. The c.1245+1G>A mutation caused a deletion of exon 9 in the SPAST gene. Prenatal genetic diagnosis showed that fetus did not carry the ALT1 c.1204T>G mutation. Follow-up was maintained for 5 years, and the negative result was confirmed by evidence of a healthy growing boy. We identified two novel mutations and two previously reported mutations in SPAST and ATL1, respectively. The family with the ATL1 c.1204T>G mutation exhibited male-lethality, female infancy-onset, and pseudo- X-linked dominant transmission, which had never been previously reported for HSP. Characteristic facial features were also noticed. The boy on whom prenatal gene diagnosis was performed is healthy and without unusual facies, suggesting that the c.1204T>G mutation might be related to these features. The results extend the genetic spectrum of HSP and suggest that linkage analysis remains a powerful tool in gene discovery studies.