[Effects of protein disulfide isomerase on hyperglycemia and hypoxia/reoxygenation injury in H9c2 cardiomyocytes].

Objective: To explore the effect of protein disulfide isomerase (PDI) in diabetic ischemic heart disease. Methods: We established an in vitro model of high glucose and hypoxia/reoxygenation in H9c2 rat myocardial cells. Cultured cells were divided into four groups: Control, high glucose (HG), hypoxia/reoxygenation (H/R) and HG+H/R. Changes in PDI expression mediated by PDI adenovirus(Ad-PDI) infection and siRNA(PDI-siRNA) transfection in myocardial cells were observed by inverted fluorescence microscopy. We also measured lactate dehydrogenase(LDH) activity and malondialdehyde(MDA) and high molecular weight(HMW)-APN concentrations. PDI, APN, cleaved caspase-3, and glucose regulated protein 78 (Grp78) protein expression were detected. Results: PDI expression was significantly decreased in the HG, H/R and HG+H/R groups compared to the Control group; however, LDH activity[(179.7±10.4) U/L、(218.4±18.4) U/L、(328.2±5.3) U/L vs (91.0±11.0) U/L], MDA concentration[(7.0±0.4) µmol/L、(10.0±1.0) µmol/L、(11.7±1.0) µmol/L vs (4.2±1.8) µmol/L], cleaved caspase-3, and Grp78 expression were increased. Interestingly, APN and HMW-APN expression were decreased [(2.01±0.21) µg/L、(1.64±0.27) µg/L、(1.20±0.14) µg/L vs (2.62±0.12) µg/L, all P<0.05]. Over expression of PDI attenuated high glucose and hypoxia/reoxygenation induced apoptosis and oxidative stress in H9c2 cardiomyocytes(all P<0.05), and simultaneously increased APN and HMW-APN expression [(2.86±0.03) µg/L vs (3.03±0.10) µg/L、(2.06±0.05) µg/L vs (2.31±0.06) µg/L、(1.83±0.07) µg/L vs (1.96±0.11) µg/L、(1.20±0.06) µg/L vs (1.39±0.09) µg/L]. PDI-siRNA transfection increased LDH activity, MDA concentration, and cleaved caspase-3 and Grp78 expression, and decreased APN and HMW-APN expression [(0.75±0.09) µg/L vs (0.59±0.09) µg/L、(0.62±0.04) µg/L vs (0.53±0.05) µg/L、(0.55±0.14) µg/L vs (0.51±0.12) µg/L、(0.48±0.12) µg/L vs (0.35±0.08) µg/L] in response to different treatments in cultured H9c2 cardiomyocytes (all P<0.05). Conclusion: PDI may regulate the expression of APN and HMW-APN, and play an important role in the function of diabetic ischemia-reperfusion cardiomyocytes.

[Analysis of pregnancy outcome of single and double blastocysts in the freeze-thaw cycle].

Objective: To compare the clinical outcomes of one and two blastocysts in the freeze-thaw transplantation cycle. Methods: Totally 3 675 cycles of frozen thawed blastocyst transplantation in Reproductive Medical Center of the Second Nanning People's Hospital from January 2012 to December 2016 were analyzed retrospectively. According to the quantity and quality of transferred blastocysts, all the patient were divided into two groups: (1) one embryo group, including the single excellent group (one high quality blastocyst) and the single non excellent group (one non high quality blastocyst); (2) two embryo groups, including the double excellent group (two high quality blastocysts), the one excellent and one non excellent group (one high quality blastocyst+one non high quality blastocyst), and the two non excellent group (two non high quality blastocysts were transplanted). Then the patients were divided into subgroups according to their ages: less than 35 years old, 35-40 years old and over 40 years old. On this basis, the implantation rate, clinical pregnancy rate, multiple birth rate and live birth rate were compared. Results: (1) The implantation rate, clinical pregnancy rate, multiple birth rate, preterm birth rate and live birth rate were all significantly increased, while the abortion rate was significantly reduced in the double blastocyst group (all P<0.05). (2) In the group of<35 years old, the rates of multiple birth and preterm birth in the double blastocyst group were significantly higher than those in the single optimal group (P<0.01). (3) In the 35-40 years old group, the clinical pregnancy rate, multiple birth rate and live birth rate of the double excellent group were significantly higher than those of the single excellent group (P<0.01); while the clinical pregnancy rate and live birth rate of the one excellent and one non excellent group and the double non excellent group were not significantly different from those of the single excellent group (P>0.05), but the multiple birth rate and preterm birth rate were significantly increased (P<0.01). The clinical pregnancy rate, live birth rate and multiple birth rate of double non optimal group were significantly higher than those of single non optimal group (P<0.01). (4) In the group>40 years old, there were no significant differences in clinical pregnancy rate and live birth rate between the two groups (P>0.05). There were no significant differences in implantation rate, clinical pregnancy rate and live birth rate between double non optimal group and single non optimal group (P>0.05). Conclusion: No matter the age of the patients, if the couple have high quality blastocysts, we should give priority to single high quality blastocyst transplantation; even if they have no high quality blastocysts, we should also consider single blastocyst transplantation, in order to reduce the risk of multiple pregnancy and improve the cumulative live birth rate, so as to improve the pregnancy outcome.

[Study on the related factors of esophageal cancer and precancerous lesions in rural residents aged 40-69 years in Shandong Province].

Objective: To analyze the related factors of esophageal squamous cell carcinoma and precancerous lesions among residents aged 40-69 years old in rural areas of Shandong Province. Methods: In October 2018, 300 villages in 13 counties of the Shandong upper gastrointestinal cancerearly diagnosis and treatment projectin 2017 were selected as research areas, and 30 400 residents aged 40-69 were recruited in this study. The demographic characteristics, health status and lifestyle information were collected through the questionnaire survey, and endoscope iodine staining and indicative biopsy methods were used for cancer screening among eligible people.The multivariate logistic regression model was used to analyze the risk factors for esophageal cancer and precancerous lesions. Results: The subjects in this study were (56.42±7.24) years old, including 13 193 males (43.40%).There were 936 cases of esophageal cancer and precancerous lesions (3.08%), including 521 males and 415 females.Compared with women, 40-49 years old, high level education, drinking tap water, regular intake of meat, eggs and milk, and family average annual income more than 30 000 RMB, men (OR=1.90, 95%CI: 1.65-2.19), 60-69 years old (OR=5.28, 95%CI: 4.11-7.30), primary school education or below (OR=1.50, 95%CI: 1.20-1.89), drinking groundwater (OR=1.71, 95%CI: 1.38-2.13), never eating meat, eggs and milk (OR=1.48, 95%CI: 1.22-1.80), and family average annual income less than 30 000 RMB (OR=1.41, 95%CI: 1.16-1.70) would increase the risk of esophageal cancer and precancerous lesions. Conclusion: The gender, age, educational level, annual household income, drinking water source, the frequency of eating meat, egg and milk were related to the occurrence of esophageal cancer and precancerous lesions among 40-69 years old residents in rural areas of Shandong Province.

[Quantification of optical coherence tomographic angiography images in type 2 diabetic patients].

Objective: To quantify optical coherence tomographic angiography (OCTA) images and to explore potential clinical values of each parameter in diabetic patients. Methods: A case-control and cross-sectional study of diabetic (129 eyes) and healthy (32 eyes) subjects was conducted. 3-mm OCTA scan, fundus photography and best corrected visual acuity measurement were performed. Image-pro plus was used to calculate microvascular and foveal avascular indices in nonsegmented and segmented OCTA images. Intraclass correlation coefficient and relative standard deviation were used to examine the reliability, reproducibility and accuracy. Correlation of each parameter was calculated, and so was the area under the receiver operating characteristic curve. Results: Vessel density (VD) of diabetic patients measured in the nonsegmented retinal layer (RL) and superficial retinal layer (SRL) was 49.146%±6.097%, 44.038%±5.641%, respectively, significantly lower than healthy subjects 52.212%±6.250%, 46.698%±5.417% (t=2.534, 2.405; P=0.012, 0.017), and vessel length (VL) was (19.905±2.285)mm, (17.596±2.149)mm, (14.479±2.091)mm in the three layers, which was significantly shorter than that in the healthy control group (21.037±2.185)mm, (18.739±1.994)mm, (15.343±2.266)mm (t=2.529, 2.731, 2.059; P=0.012, 0.007, 0.041). Compared with non-diabetes, foveal avascular zone (FAZ) in diabetes was (0.441±0.167)mm(2), larger compared to health eyes 0.352±0.109mm(2) (t=-2.831, P=0.005). FAZ demonstrated a negative correlation with VD and VL in RL or SRL(r=-0.227, -0.338; P<0.05) (r=-0.241, -0.332; P<0.05), while best corrected visual acuity showed a negative correlation with VL in each layer (r=-0.225, -0.201, -0.250; P=0.01, 0.022, 0.004), and VD in DRL (r=-0.197, P=0.026). All parameters showed high reproducibility between graders(intraclass correlation coefficient>0.965), while relative standard deviation was greater than 3.049%. AR had the highest area under the receiver operating characteristic curve (0.737), compared with AI (0.724). Conclusions: Our study demonstrates good reliability, reproducibility and accuracy of quantification of OCTA images and reveals changes of VD, VL and FAZ in diabetes, which may occur earlier than vision impairment. AR and AL may have great values in prompting early diabetic retinopathy in diabetic patients. (Chin J Ophthalmol, 2019, 55:273-279).

[An association of ulcerative colitis with tumor necrosis factor superfamily member 15 gene polymorphisms in Chinese patients].

Objective: To explore the relationship between ulcerative colitis (UC) susceptibility and tumor necrosis factor superfamily member (TNFSF) 15 gene polymorphisms and haplotypes in Han nationality in Zhejiang province of China. Methods: A total of 408 UC patients and 574 healthy controls were recruited in this study. Three single nucleotide polymorphisms of TNFSF15 (rs3810936, rs4263839, rs4979462) were examined by improved multiple ligase detection reaction (iMLDR) technique. Analyses of linkage disequilibrium (LD) and haplotype were performed by Haploview 4.2 software in all study subjects. Results: The variant allele A and genotype (GA+AA) of rs4263839 were less frequent in UC patients than in controls (45.34% vs. 50.17%, P=0.035;68.38% vs. 76.66%, P=0.004). According to the severity and location of disease, UC patients were divided into different subgroups. After multiple comparison correction(α=0.012 5), the frequencies of variant allele A and genotype (GA+AA) of rs4263839 were lower in patients with severe UC than in the controls (37.69% vs. 50.17%, P=0.007; 60.00% vs. 76.66%, P=0.004). Similar findings were also drawn for patients with extensive colitis in contrast with the controls (42.22% vs. 50.17%, P=0.009; 63.33% vs. 76.66%, P<0.001). Furthermore, the haplotype analysis indicated that three SNPs above were in a strong LD. The frequency of haplotype TAC was lower in UC patients than in the controls(40.83% vs. 46.04%, P=0.023). Also it was less prevalent in patients with severe UC and patients with extensive colitis when compared with controls respectively (33.38% vs. 46.04%, P=0.005;37.22% vs. 46.04%, P=0.003). Conclusions: TNFSF15 (rs4263839) variation might not only reduce the risk of UC, but also affect the severity and lesion location of UC. The haplotype TAC formed by rs3810936, rs4263839 and rs4979462 might be related to a lower risk of UC, especially in patients with severe colitis or patients with extensive colitis.

[Association of Crohn's disease with aryl hydrocarbon receptor gene polymorphisms and haplotypes].

Objective: To explore the relationship of Crohn's disease (CD) susceptibility to aryl hydrocarbon receptor (AhR) polymorphisms and haplotypes in Han population in Wenzhou city, China. Methods: A total of 310 CD patients and 573 age- and sex-matched healthy controls were enrolled in our study. Three single nucleotide polymorphisms (SNPs) of AhR(rs10249788,rs2158041,rs2066853) were determined by the improved multiple ligase detection reaction technique. Unconditional logistic regression analyses was applied to analyze the allelic and genotypic differences of each SNP between CD patients and controls, as well as their influence on the clinicopathologic characteristics in CD patients. Analyses of linkage disequilibrium and haplotype were performed by Haploview 4.2 software in all study subjects. Results: Compared with the controls, the variant allele (T) and genotype (CT+TT) of (rs2158041) were evidently decreased among CD patients (19.52% vs. 25.04%, P=0.009; 34.19% vs. 44.68%, P=0.003). According to "the Montreal Classification Standards" , CD patients were divided into different subgroups. The variant allele (T) and genotype (CT+TT) of (rs2158041) were significantly lower in patients with terminal ileum CD than in controls (16.79% vs. 25.04%, P=0.005; 28.24% vs. 44.68%, P=0.001). Similar conclusions were also drawn in patients with constricting disease when compared with the controls (15.20% vs. 25.04%, P=0.003; 28.43% vs. 44.68%, P=0.003). The three SNPs above were shown to be in a linkage disequilibrium. Compared with the controls respectively, the frequency of haplotype (CCG) was increased in CD patients (44.73% vs. 39.60%, P=0.039), whereas that of haplotype (CTG) was decreased (18.02% vs. 22.78%, P=0.047). Conclusions: AhR (rs2158041) variation might influence the risk as well as the location and behavior of CD. The haplotype (CCG) possibly increase the risk of CD development, whereas haplotype (CTG) might decrease it.

Promoter analysis and tissue expression of the chicken ASB15 gene.

1. This study was conducted to explore the promoter region of the chicken ASB15 gene by detecting the activities of the dual luciferase reporter gene and to assess expression profiles of the ASB15 gene in 10 different tissues from Gushi chickens. 2. Five dual luciferase reporter gene vectors were constructed and transfected into DF1 cells. The activities of recombined plasmids were measured and the core promoter was confirmed by bioinformatic analysis. Total RNA was extracted and the relative expression of the ASB15 gene was examined. 3. Data analysis indicated that the promoter was located from -955 to -212 bp. Results showed that the chicken ASB15 gene was expressed in heart, breast muscle and leg muscle. 4. This study has confirmed the promoter region and the expression profile of the chicken ASB15 gene, which provides a foundation for further exploring its transcriptional regulation and function.

Morphometrics and molecular analysis of the free-living nematode, Belondira bagongshanensis n. sp. (Dorylaimida, Belondiridae), from China.

A new species, Belondira bagongshanensis n. sp., extracted from soil under grass in Bagongshan Forest Park, Anhui Province, China, is described and illustrated. The new species is mainly characterized by a body length 1.6-2.1 mm; cephalic framework moderately sclerotized; odontostyle robust with a distinct lumen; anterior part of pharynx with a distinct fusiform swelling, bearing distinct sclerotized valve plates and basal expansion occupying about three-fifths of the total neck length; female genital system mono-opisthodelphic and anterior branch 0.8-1.6 times the corresponding body diameter; tail short and hemispheroid; male spicules 36-49 µm long and two widely spaced ventromedian supplements. The new species is close to B. wajahati, B. golden and B. eugeniae in possessing a short tail, a robust odontostyle with a distinct aperture and a moderately sclerotized cephalic framework. New sequences of 18S rDNA and the D2-D3 region of 28S rDNA of the new species were also identified.

Morphology of Labronemella major n. sp. (Nematoda: Dorylaimida), a soil-dwelling nematode from China, including a revised key to species of the genus.

One new species from Qinghai Province, China, Labronemella major n. sp., is described. The new species is characterized by a body length of 3.03-3.34 mm; lip region wide, offset by a distinct depression, disc-like with six separated inner liplets; amphid fovea funnel-shaped, distinctly bulged on body surface in scanning electron micrographs; odontostyle long (35-39 µm) with distinct lumen, aperture about 39-47% of its length; odontophore rod-like and long; guiding ring double; pharyngeal basal expansion about half of the total neck length; uterus relatively long and tripartite; vulva transverse and sclerotized; spicules 81-90 µm long; ventromedial supplements 19-23; tail short, rounded to conoid. It can be differentiated from all other species of the genus by its relatively longer body, odontostyle and spicules, and wider lip region. Due to the lip region being offset by a deep constriction, and the long (three or more times the body diameter at mid-body) tripartite uterus, the new species is close to Labronemella czernowitzensis (Micoletzky, 1922) Andrássy, 2002 and Labronemella labiata Andrássy, 1985. An improved key to the genus including the new species is provided.

Morphology and molecular analysis of Paratylenchus nanjingensis n. sp. (Nematoda: Paratylenchinae) from the rhizosphere soil of Pinus massoniana in China.

Paratylenchus nanjingensis n. sp. was obtained from Nanjing, Jiangsu Province, China. This new species is characterized by having a female with a slender, vermiform body (243-279 µm), head with distinct submedian lobes, slender and long stylet (64-68 µm), anchor-shaped stylet knobs, excretory pore anterior to the level of the stylet knobs, small lateral vulval flaps and lateral field with four lines; and male with more distinct body annuli, stylet lacking and pharynx degenerate. The internal transcribed spacer sequences of ribosomal RNA (ITS rRNA) gene of the new species were amplified and sequenced in this study. The phylogenetic relationships of the new species with other Paratylenchus species using the ITS rRNA gene sequences are given.

Morphology and morphometrics of Heterodorus qinghaiensis n. sp. (Dorylaimida, Nordiidae) from soil samples in China.

One new species of the family Nordiidae Jairajpuri & Siddiqi, 1964 from the enclosure grassland of Qinghai Province, China, Heterodorus qinghaiensis n. sp., is described and illustrated. The new species is characterized by the slender body, 1.29-1.46 mm in length; the granular lateral chord with numerous large depression plaques throughout its entirety; the lip region offset by a distinct depression; amphid goblet-shaped with aperture about half to two-thirds of corresponding body diameter; odontostyle 11-13 µm long; rod-like odontophore without basal flanges; pharyngeal basal expansion about one-third of the total neck length; didelphic genital system containing sperm; ovaries generally not reaching the sphincter level; vulva transversed and sclerotized; female tail conoid with round terminus; 3-5 spaced ventromedial supplements and spicule 32-41 µm long. It is close to H. liangi (Ahmad, Wu & Shaheen, 2002) Andrássy, 2009, H. brevidentatus (Thorne, 1939) Andrássy, 2009, H. monticola Andrássy, 2011, H. morgensis (Loof, 1988) Andrássy, 2009 and H. meghalayensis (Mushtaq, Baniyamuddin & Ahmad, 2007) Andrássy, 2009 in having inconspicuous or no lateral body pores, smaller odontostyle and ventrally curved tail.

Effect of atorvastatin on diabetic rat endothelial cells and retinal lesions.

We investigated the effect of atorvastatin on vascular endothelial growth inhibitor (VEGI) expression in rats with diabetic retinopathy. Wistar rats were divided into a blank group and diabetic model group, which was further randomly divided into treatment and control groups. Rats in the treatment group received 10 mg/kg atorvastatin daily, while rats in the blank and control groups received normal saline. Rats were randomly euthanized at 3 or 6 months. Immunohistochemical staining was used to determine changes in VEGI and vascular endothelial growth factor, interleukin-4, and tumor necrosis factor α levels in rats with diabetic retinopathy. Survival rate in the treatment group was 84% (63/75) after 6 months, which was significantly higher than that in the control group (P < 0.05); rats in the control group showed the lowest survival rate. Survival in the treatment group was higher than that in the control group but not significant compared with the blank group after 3 months. VEGI, vascular endothelial growth factor, tumor necrosis factor α, and interluekin-4 expression was lower than that in the control group, but higher than the blank group after 3 months. The expression of each factor decreased to the blank group level in the treatment group and was significantly lower than that in the control group after 6 months (P < 0.05). Expression in control and blank groups was similar at 3 and 6 months. Atorvastatin can inhibit VEGI and vascular endothelial growth factor expression to protect rats from diabetic retinopathy.

[Mitochondrial pyruvate carrier deficiency: 3 cases report and literature review].

Objective: To analyze the clinical and genetic features of patients with mitochondrial pyruvate carrier deficiency (MPYCD). Methods: This was a case series research. The clinical data, genetic characteristics, and glutamine treatment efficacy of 3 patients diagnosed with MPYCD at the Department of Neurology, Beijing Children's Hospital, Capital Medical University and Department of Pediatrics, Guizhou Provincial People's Hospital, from August 2019 to June 2023 were retrospectively collected. A literature search with "MPC1 gene" "MPC2 gene and" "mitochondrial pyruvate carrier deficiency" as keywords was conducted at the Wanfang Data Knowledge Service Platform, China National Knowledge Infrastructure (CNKI) and PubMed (up to June 2023). Clinical and genetic characteristics of patients with MPYCD were summarized. Results: Case 1 was a 3 years and 11 months old boy, while case 2 was a 4 years and 10 months old boy and case 3 was an 8 years and 9 months old girl. Case 2 and case 3 were siblings from one consanguineous family. All 3 patients presented with general developmental delay, growth failure and elevated serum lactate. Cranial magnetic resonance imaging (MRI) showed subtle bilateral symmetrical T2 signal hyperintensity in basal ganglia and thalamus in case 1, but normal in case 2 and 3. Trio-WES revealed case 1 harboring compound heterozygous missense variants c.208G>A (p.Ala70Thr) and c.290G>A (p.Arg97Gln) in MPC1 gene, while case 2 and 3 revealed a homozygous variant c.290G>A (p.Arg97Gln) in the same gene. All 3 cases were diagnosecl as MPYCD. Clinical symptoms including motor ability, cognition and activity endurance were improved in these 3 patients after taking glutamine for 2 years. A total of 5 articles published in English were reviewed, and no Chinese literature was found. Including these 3 cases, 15 cases were enrolled for analysis. Eleven patients carried MPC1 gene variants and 4 cases carried MPC2 gene variants. Except for 3 cases died during prenatal period, 9 of 12 enrolled born cases were onset before 6 months old. The most common clinical symptoms were mental and motor general developmental delay, microcephaly, growth failure and hypotonia. All patients had elevated blood lactate and pyruvate, but the ratio of lactate/pyruvate was normal. Seven patients performed cranial MRI, 3 exhibited non-specific changes, 2 showed bilateral symmetrical T2 signal hyperintensity in basal ganglia and thalamus, and 3 were normal. A total of 5 MPC1 gene missense variants and 2 MPC2 gene variants were identified in 15 cases. Conclusions: Onset age of patients with MPYCD is usually within 6 months. The main clinical characteristics are developmental delay, microcephaly and growth failure, accompanied by increased serum lactate and pyruvate. Glutamine supplement could lead to clinical improvements.

First Report of Root-Knot Nematode Meloidogyne arenaria Infesting Roots of Anubias barteri in Guangdong, China.

Anubias spp. are very popular aquatic plants that are native to Africa. Anubias barteri is a broadleaf Anubias and is a popular choice for aquariums because of its robust size. In China, broadleaf Anubias is usually planted in sand, with sponges around the rhizome, in small plastic baskets in aquatic plant nurseries. In December 2010, a survey of phytoparasitic nematodes was conducted in the nurseries in Guangzhou, Guangdong Province. Many second-stage juveniles of a Meloidogyne sp. were detected from the roots of A. barteri, but no root galls were found. To identify these juveniles, molecular identification was performed with PCR. The DNA sequence between CO II and l-rRNA of the mitochondrial gene of single juveniles was amplified with universal primers of Meloidogyne, #C2F3 (5'-GGTCAATGTTCAGAAATTTGTGG-3') and #1108 (5'-TACCTTTGACCAATCACGCT-3') (3). The amplified fragments were approximately 1.1 kb long and could not be digested with restrictive enzyme HinfI. The specific fragments were then sequenced. The blast search result revealed that the DNA sequence (GenBank Accession No. JQ446377) had 99 to 100% identity with submitted sequences of Meloidogyne arenaria (GenBank Accession Nos. EU364879, GQ266686, and AY635610). The other extracted juveniles were inoculated into sterile, potted, water spinach (Ipomoea aquatica) in the greenhouse to obtain more nematodes. After 40 days, root galls and female egg masses were clearly observed, and biochemical, molecular, and morphological identifications were conducted. Isoenzyme phenotype (esterase and malate dehydrogenase (MDH) patterns) and the perineal pattern of several gravid females were the same as M. arenaria (1,2), and PCR amplification of single juveniles produced identical fragments as previously found. Single egg masses were collected and juveniles were hatched out and inoculated onto 10 nematode-free plants of A. barteri in a greenhouse. After 40 days, roots of A. barteri exhibited inconspicuous small galls, and the same identification procedures were conducted as mentioned previously. Isoenzyme phenotypes, perineal patterns of adult females, and amplified fragments of single juveniles were identical to those of M. arenaria. M. arenaria is one of the most important root-knot nematodes and causes great losses in many crops around the world (2). To our knowledge, this is the first record of M. arenaria parasitizing aquatic plants of broadleaf Anubias in China and elsewhere, and A. barteri is a new host of M. arenaria. The economic importance of this nematode to A. barteri production is currently unknown. However, because A. barteri is a commercial aquatic plant, more attention should be given by producers to prevent this nematode from becoming an important pathogen. In addition, this finding is very helpful for relevant plant nematode quarantine work. References: (1) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 17:1, 1985. (2) R. N. Perry et al. Root-Knot Nematodes. CABI. Wallingford, UK, 2009. (3) T. O. Powers and T. S. Harris. J. Nematol. 25:1, 1993.

Prognostic value of LncRNA-HOTAIR for patients with hepatocellular carcinoma: a meta-analysis.

OBJECTIVE: This study aims at evaluating the prognostic value of LncRNA-HOTAIR for patients with hepatocellular carcinoma (HCC). MATERIALS AND METHODS: A comprehensive databases and literature search was performed on PubMed, EMBASE, Web of Science, the Cochrane Library, CNKI, CBM, Wanfang, and VIP database up to the end of February 2022, for published studies on the connection of HCC and HOTAIR. STATA 12.0 software was used for the meta-analysis. RESULTS: Eight studies with 412 patients were selected to be entered in the meta-analysis. We found that high expression of HOTAIR was associated with III+IV tumor stage (HR=2.31, 95% CI:1.32, 4.01), and it was not associated with age (HR=0.86, 95% CI:0.55, 1.34), gender (HR=0.91, 95% CI:0.55, 1.46), tumor number (HR=1.58, 95% CI:0.72, 3.48), tumor size (HR=1.54, 95% CI:0.96, 2.49), lymph node metastasis (HR=0.66, 95% CI:0.38, 1.15), AFP (HR=0.81, 95% CI:0.46, 1.42), cirrhosis (HR=1.34, 95% CI:0.75, 2.41), or portal invasion (HR=1.76, 95% CI:0.83, 3.72). A high expression level of HOTAIR was associated with a poorer OS (HR=3.12, 95% CI:1.31-7.43, p=0.010) and RFS (HR=1.67, 95% CI:1.23-2.26, p=0.010) for patients with HCC. CONCLUSIONS: A high expression level of HOTAIR was associated with III+IV tumor stage. Our meta-analysis clearly supports the prognostic value of HOTAIR to predict unfavorable prognostic outcomes for patients with HCC.

[Clinical significance and pathogenesis analysis of heterogeneous nuclear ribonucleoprotein U in acute myeloid leukemia].

Objective: To investigate the clinical significance and pathogenesis of heterogeneous nuclear ribonucleoprotein U (hnRNP U) in acute myeloid leukemia (AML) . Methods: The expression of hnRNP U, an RNA binding protein, in patients with AML and healthy controls was compared based on the Gene Expression Profiling Interactive Analysis database and the data of the center. The Beat AML Dataset (n=158) was downloaded from the cBioPortal database. The hnRNP U expression level was divided into the high-expression group (n=89) and low-expression group (n=69) , and patients' clinical characteristics were compared. The effect of hnRNP U on the biological behavior of human AML cell lines was studied by Cell Counting Kit-8 assay to detect cell proliferation. Annexin Ⅴ-APC/7-AAD antibodies were used to detect cell apoptosis. DNA content (PI staining) was quantitatively analyzed to detect cell cycle changes, and colony formation experiments were performed to detect cell cloning formation ability after hnRNP U knockdown in Kasumi-1 and MOLM-13 cells. To study the effect of hnRNP U knockdown on the DNA damage response (DDR) pathway proteins of cleaved-PARP, immunoblot analysis using p-H2A.X was conducted. Results: ①Pan-cancer analysis showed that hnRNP U was highly expressed in patients with AML, and the expression level of hnRNP U mRNA in peripheral blood mononuclear cells was significantly higher in patients with AML than in healthy controls (0.0315±0.0042 vs 0.0195±0.0006, respectively, P<0.01) . ②The age of onset was 56 (2-87) years in the high-expression group and 65 (8-85) years in the low-expression group (t=-2.681, P=0.007) . Moreover, the high-expression group had a higher proportion of combined FLT3 mutations than the low-expression group (χ(2)=4.069, P=0.044) . ③Compared with the negative control, hnRNP U knockdown inhibited the proliferation (P<0.001 and P<0.001) , promoted the apoptosis (P<0.01 and P<0.001) , decreased the colony formation ability (P<0.001 and P<0.001) , and arrested the cell cycles in the G(2)/M phase (P<0.05 and P<0.01) of Kasumi-1 and MOLM-13 cells, respectively. ④hnRNP U knockdown could increase the protein expression of cleaved-PARP and p-H2A.X on the DDR pathway. Conclusion: hnRNP U is highly expressed in AML, and hnRNP U knockdown can inhibit the occurrence and development of AML possibly through the activation of the DDR pathway.

[Enzyme immunoassay for the determination of hexestrol in meat].

An indirect competitive enzyme-linked immunosorbent assay (ELISA) of hexestrol (HES), an antibiotic forbidden for use in livestock farming, has been developed. Conditions of ELISA have been optimized by varying the concentrations of the coating conjugate (HES-ovalbumin), anti-HES antiserum, casein, and Tween 20. In the absence of Tween 20 in the reaction mixture, the detection limit (IC10) equaled 0.01 ng/ml, IC50 equaled 0.17 ng/ml, and the working range (IC20-IC80) equaled 0.03-0.86 ng/ml, while, in the presence of 0.05% Tween 20, these values equaled 0.05 ng/ml, 2.9 ng/ml, and 0.26-32.0 ng/ml, respectively. Standard deviation of the analysis results did not exceed 5.4%. If ELISA was performed in the absence of detergents, the recovery value upon HES determination in spiked beef samples ranged from 74 to 147%.

Size-dependent survival of brook trout Salvelinus fontinalis in summer: effects of water temperature and stream flow.

A 5 year individual-based data set was used to estimate size-specific survival rates in a wild brook trout Salvelinus fontinalis population in a stream network encompassing a mainstem and three tributaries (1.5-6 m wetted width), western Massachusetts, U.S.A. The relationships between survival in summer and temperature and flow metrics derived from continuous monitoring data were then tested. Increased summer temperatures significantly reduced summer survival rates for S. fontinalis in almost all size classes in all four sites throughout the network. In contrast, extreme low summer flows reduced survival of large fish, but only in small tributaries, and had no significant effects on fish in smaller size classes in any location. These results provide direct evidence of a link between season-specific survival and environmental factors likely to be affected by climate change and have important consequences for the management of both habitats and populations.

Protocol optimization for long-term liquid storage of goat semen in a chemically defined extender.

A specific problem in the preservation of goat semen has been the detrimental effect of seminal plasma on the viability of spermatozoa in extenders containing egg yolk or milk. The use of chemically defined extenders will have obvious advantages in liquid storage of buck semen. Our previous study showed that the self-made mZAP extender performed better than commercial extenders, and maintained a sperm motility of 34% for 9 days and a fertilizing potential for successful pregnancies for 7 days. The aim of this study was to extend the viability and fertilizing potential of liquid-stored goat spermatozoa by optimizing procedures for semen processing and storage in the mZAP extender. Semen samples collected from five goat bucks of the Lubei White and Boer breeds were diluted with the extender, cooled and stored at 5 degrees C. Stored semen was evaluated for sperm viability parameters, every 48 h of storage. Data from three ejaculates of different bucks were analysed for each treatment. The percentage data were arcsine-transformed before being analysed with anova and Duncan's multiple comparison test. While cooling at the rate of 0.1-0.25 degrees C/min did not affect sperm viability parameters, doing so at the rate of 0.6 degrees C/min from 30 to 15 degrees C reduced goat sperm motility and membrane integrity. Sperm motility and membrane integrity were significantly higher in semen coated with the extender containing 20% egg yolk than in non-coated semen. Sperm motility, membrane integrity and acrosomal intactness were significantly higher when coated semen was 21-fold diluted than when it was 11- or 51-fold diluted and when extender was renewed at 48-h intervals than when it was not renewed during storage. When goat semen coated with the egg yolk-containing extender was 21-fold diluted, cooled at the rate of 0.07-0.25 degrees C/min, stored at 5 degrees C and the extender renewed every 48 h, a sperm motility of 48% was maintained for 13 days, and an in vitro-fertilizing potential similar to that of fresh semen was maintained for 11 days.

Liquid storage of goat semen in chemically defined extenders.

The suitability of certain commercial and self-made chemically defined extenders for liquid storage of goat semen was tested and the effects of storage temperatures, dilution rates and sperm washing and pH of extenders on the goat sperm during liquid storage were observed. Semen was collected from nine goat bucks of the Lubei White and Boer breeds using an artificial vagina. Each ejaculate after initial evaluation was diluted with a specific extender, cooled and stored at a desired temperature. Stored semen was evaluated for sperm motility and other parameters every 24 or 48 h of storage. The ranking order of the existing milk- and yolk-free extenders in sustaining goat sperm motility was Androhep > Zorlesco > Beltsville thawing solution > the Tris-glucose medium. The new extender (mZA) which was formulated based on Zorlesco and Androhep was more suitable for goat sperm than Androhep. The mZAP extender with Bovine Serum Albumin (BSA) replaced with polyvinyl alcohol (PVA) worked as efficiently as the mZA in maintaining sperm motility, membrane integrity, acrosome intactness and capacitation status. Goat sperm motility was best maintained at 5 degrees C during liquid preservation, but decreased significantly as the temperature increased. When semen was sixfold diluted, sperm motility was maintained longer (p < 0.05) after centrifugation, but sperm motility did not differ between the centrifuged and non-centrifuged groups when semen was 11-fold diluted. When the extender pH was adjusted from 6.6 to 6.04, the efficiency increased significantly in both Androhep and mZAP. A forward sperm motility of 34% was maintained for 9 days when buck semen was 11-fold diluted and stored at 5 degrees C in mZAP, with pH adjusted to 6.04. It is concluded that for liquid storage of buck semen, the mZA extender was more suitable than other extenders; BSA can be replaced with PVA in mZA; centrifugation to remove seminal plasma can be omitted by adequate dilution; and the storage temperature and pH of extenders affected sperm motility significantly.