[Cloning and expression analysis of U6 promoters in Panax quinquefolius].

The U6 promoter is an important element driving sgRNA transcription in the CRISPR/Cas9 system. Seven PqU6 promo-ter sequences were cloned from the gDNA of Panax quinquefolium, and the transcriptional activation ability of the seven promoters was studied. In this study, seven PqU6 promoter sequences with a length of about 1 300 bp were cloned from the adventitious roots of P. quinquefolium cultivated for 5 weeks. Bioinformatics tools were used to analyze the sequence characteristics of PqU6 promoters, and the fusion expression vectors of GUS gene driven by PqU6-P were constructed. Tobacco leaves were transformed by Agrobacterium tumefaciens-mediated method for activity detection. The seven PqU6 promoters were truncated from the 5'-end to reach 283, 287, 279, 289, 295, 289, and 283 bp, respectively. The vectors for detection of promoter activity were constructed with GUS as a reported gene and used to transform P. quinquefolium callus and tobacco leaves. The results showed that seven PqU6 promoter sequences(PqU6-1P to PqU6-7P) were cloned from the gDNA of P. quinquefolium, with the length ranged from 1 246 bp to 1 308 bp. Sequence comparison results showed that the seven PqU6 promoter sequences and the AtU6-P promoter all had USE and TATA boxes, which are essential elements affecting the transcriptional activity of the U6 promoter. The results of GUS staining and enzyme activity test showed that all the seven PqU6 promoters had transcriptional activity. The PqU6-7P with a length of 1 269 bp had the highest transcriptional activity, 1.31 times that of the positive control P-35S. When the seven PqU6 promoters were truncated from the 5'-end(PqU6-1PA to PqU6-7PA), their transcriptional activities were different in tobacco leaves and P. quinquefolium callus. The transcriptional activity of PqU6-7PA promoter(283 bp) was 1.59 times that of AtU6-P promoter(292 bp) when the recipient material was P. quinquefolium callus. The findings provide more ideal endogenous U6 promoters for CRISPR/Cas9 technology in ginseng and other medicinal plants.

Myelin sheath structure and regeneration in peripheral nerve injury repair.

Observing the structure and regeneration of the myelin sheath in peripheral nerves following injury and during repair would help in understanding the pathogenesis and treatment of neurological diseases caused by an abnormal myelin sheath. In the present study, transmission electron microscopy, immunofluorescence staining, and transcriptome analyses were used to investigate the structure and regeneration of the myelin sheath after end-to-end anastomosis, autologous nerve transplantation, and nerve tube transplantation in a rat model of sciatic nerve injury, with normal optic nerve, oculomotor nerve, sciatic nerve, and Schwann cells used as controls. The results suggested that the double-bilayer was the structural unit that constituted the myelin sheath. The major feature during regeneration was the compaction of the myelin sheath, wherein the distance between the 2 layers of cell membrane in the double-bilayer became shorter and the adjacent double-bilayers tightly closed together and formed the major dense line. The expression level of myelin basic protein was positively correlated with the formation of the major dense line, and the compacted myelin sheath could not be formed without the anchoring of the lipophilin particles to the myelin sheath.

Characterization and expression analyses of somatolactin-α and -ß genes in rare minnows (Gobiocypris rarus) following waterborne cadmium exposure.

Using reverse transcription-polymerase chain reaction (RT-PCR) and RACE (rapid amplification of cDNA ends), somatolactin-α (rmSLα) and -ß (rmSLß) were identified from the pituitary gland of rare minnows (Gobiocypris rarus). The full-length cDNAs of these two genes were 1288 and 801 bp, encoding prepeptides of 250 and 228 amino acids residues, respectively. rmSLß can be detected in the brain (including the pituitary), ovary, testis, and gill, while rmSLα was mainly expressed in the brain. On the other hand, rmSLα was expressed in all the fetal developmental stages; however, rmSLß can just be detected in the stages since from 14 h post-fertilization (hpf). After exposure to acute waterborne cadmium (Cd), rmSLα was distinctly upregulated in juvenile rare minnows at all detected time points, from 24 to 96 h and 10 days, while rmSLß was significantly altered only in 96 h or 10-day treatment groups. As for adults, acute Cd exposure caused alterations of both rmSLα and rmSLß in the brain (containing the pituitary) at the 24 h; subchronic waterborne Cd treatment led to upregulation of rmSLα, while decrease of mSLß in the brain. Alteration of rmSL transcripts following waterborne Cd exposure further confirmed the endocrine disruption of this heavy metal. Besides, exposure to as low as 5 µg/L Cd caused alteration of rmSLα, which suggested that rmSLα might be a potential biomarker for risk assessment of aquatic Cd.

[Analysis on the application of three methods for malaria diagnosis].

Objective: To test the usage of microscopic examination, antigen detection(rapid dignostic test, RDT) and nucleic acid test(PCR) for detection of malaria cases. Methods: The blood test results for malaria and suspected malaria cases during 2012-2015 were retrospectively reviewed. Taking the confirmed cases as a gold standard, the three methods were compared in aspects of diagnosis indices, specificity of identification species, and cost effectiveness. Results: A total of 212 samples were included, each analyzed with the three methods. Based on the results of the three tests, 167(78.8%) were determined to be positive for malaria, and 45 negative (21.2%). Of the positive samples, 120(71.9%) were infected with Plasmodium falciparum,22(13.2%) with P. vivax,17(10.2%) with P. ovale, 6 (3.6%) with P. malariae, and 2(1.2%) with mixed infections. The method of PCR had the highest diagnostic efficiency (96.2%,204/212), followed by RDT (93.2%,192/206; P > 0.05 vs. PCR) and the microscopic method (88.2%,187/212; P < 0.05 vs. RDT and PCR). Similarly, the PCR method had the highest overall coincidence rate to the confirmed cases (95.3%,202/212), followed by RDT (93.2%,192/206) and microscopy (88.2%,187/212; P < 0.05 vs. PCR). As to the identification specificity among species, the PCR method(95.6%, 43/45) was superior to microscopy (91.1%, 41/45; P > 0.05 vs. PCR) and RDT (68.9%, 31/45; P < 0.05 vs. PCR). As to the identification of a particular species (P. falciparum), RDT performed best (100%,116/116), followed by PCR (93.3%,112/120) and microscopy (84.2%,101/120). Based on the comprehensive evaluation on 14 indicators including if it is a diagnostic criterion, equipment and technical requirement, diagnostic performance, time cost, and the need of technical training and promotion, we found that the RDT method had the highest score(37 of 42), while microscopy and PCR were scored 26 and 27, respectively. Conclusion: Under the falciparum malaria-dominated epidemiological situation, PCR and RDT show a higher detection efficiency, PCR and microscopy perform better in species identification, and RDT has the highest cost-effectiveness.

Acclimation temperature alters the relationship between growth and swimming performance among juvenile common carp (Cyprinus carpio).

Individual variation in growth, metabolism and swimming performance, their possible interrelationships, and the effects of temperature were investigated in 30 juvenile common carp (Cyprinus carpio) at two acclimation temperatures (15 and 25°C). We measured body mass, critical swimming speed (Ucrit), resting metabolic rate (RMR), active metabolic rate (AMR) and metabolic scope (MS) twice (28days apart) in both temperature groups. Fish acclimated to 25°C showed a 204% higher specific growth rate (SGR) than those acclimated to 15°C due to a 97% higher feeding rate (FR) and a 46% higher feed efficiency (FE). Among individuals, SGR was positively correlated with the FR and FE at both low and high temperatures. All measured variables (Ucrit, RMR and AMR) related to swimming except MS showed a high repeatability after adjusting for body mass (mass-independent). Fish acclimated to 25°C had a 40% higher Ucrit compared with 15°C acclimated fish, which was at least partially due to an improved metabolic capacity. AMR showed a 97% increase, and MS showed a 104% parallel increase with the higher acclimation temperature. Residual (mass-independent) Ucrit was positively correlated with residual RMR, AMR and MS, except for the residual RMR at high temperature. When acclimated to the lower temperature, both the residual and absolute Ucrit were negatively correlated with FR and FE and, hence, with SGR, suggesting a functional trade-off between growth and locomotion in fish acclimated to low temperatures. However, when acclimated to the higher temperature, this trade-off no longer existed; absolute Ucrit was positively correlated with SGR because individuals with rapid growth exhibited greatly increased body mass. The higher metabolic capacity at 25°C showed a positive effect on both swimming performance and growth rate (because of improved digestive efficiency) under the high-temperature condition, which we did not anticipate. Overall, these results indicate that temperature alters the relationship between growth and swimming performance of juvenile common carp. This change may be an adaptive strategy to seasonal temperature variation during their life history.

The effect of aerobic exercise and starvation on growth performance and postprandial metabolic response in juvenile southern catfish (Silurus meridionalis).

To investigate the effects of aerobic exercise and starvation on growth performance, postprandial metabolic response and their interaction in a sedentary fish species, either satiation-fed or starved juvenile southern catfish (Silurus meridionalis) were exercised at 25 °C under three water velocities, i.e., nearly still water (control), 1 body length (bl) s(-1) and 2 bl s(-1), for eight weeks. Then, the feed intake (FI), food conversion efficiency (FCE), specific growth rate (SGR), morphological parameters, resting MO2 (MO2rest) and postprandial MO2 responses of the experimental fish were measured. Exercise at a low velocity (1 bl s(-1)) showed no effect on any growth performance parameter, whereas exercise at a high velocity (2 bl s(-1)) exhibited higher FI but similar SGR due to the extra energy expenditure from swimming and consequent decreased FCE. Starvation led to a significant body mass loss, whereas the effect intensified in both exercise groups. Exercise resulted in improved cardio-respiratory capacity, as indicated by increased gill and heart indexes, whereas it exhibited no effect on resting and postprandial metabolism in S. meridionalis. The starved fish displayed significantly larger heart, gill and digestive tract indexes compared with the feeding fish, suggesting selective maintenance of cardio-respiratory and digestive function in this fish species during starvation. However, starved fish still exhibited impaired digestive performance, as evidenced by the prolonged duration and low postprandial metabolic increase, and this effect was further exacerbated in both the 1 and 2 bl s(-1) exercise groups. These data suggest the following: (1) aerobic exercise produced no improvement in growth performance but may have led to the impairment of growth under insufficient food conditions; (2) the mass of different organs and tissues responded differently to aerobic exercise and starvation due to the different physiological roles they play; and (3) aerobic exercise had no effect on the postprandial metabolic response under a "normal feeding" situation, whereas it may have resulted in the impairment of the digestive capacity when food availability was low due to the competition of energy and oxygen under unfavorable conditions in juvenile S. meridionalis.

The effects of starvation and re-feeding on growth and swimming performance of juvenile black carp (Mylopharyngodon piceus).

We investigated the effects of starvation and re-feeding on growth and swimming performance and their relationship in juvenile black carp (Mylopharyngodon piceus). We measured the specific growth rate (SGR), resting metabolic rate (RMR) and constant acceleration test speed (U CAT, the maximum swimming speed at exhaustion by constant acceleration test with 0.1667 cm s(-2) rate) in a treatment group (21 days of starvation then 21 days of re-feeding) and control group (routine feeding) (n = 20). Starvation resulted in a 17 % decrease in body mass of black carp (P < 0.05). After 21 days of re-feeding, body mass was greater than that of pre-starvation but still less than that of the control group at 42 days. During the re-feeding phase, the SGR of the treatment group was higher than that of the control group (P < 0.05). Starvation resulted in a significant decrease in the RMR and U CAT. After 21 days of re-feeding, both the RMR and U CAT recovered to the pre-starvation levels. In the control group, individual juvenile black carp displayed strong repeatability of the RMR and U CAT across the measurement periods (P ≤ 0.002). In the treatment group, RMR showed significant repeatability between pre-starvation and re-feeding (P = 0.007), but not between pre-starvation and starvation or between starvation and re-feeding. U CAT showed significant repeatability between pre-starvation and starvation (P = 0.006) and between pre-starvation and re-feeding (P = 0.001), but not between starvation and re-feeding. No correlation or only a weak correlation was found between any two variables of RMR, U CAT and SGR, whereas the increment of the U CAT (ΔU CAT) was negatively correlated with that of SGR during the starvation phase (r = -0.581, n = 20, P = 0.007) and re-feeding phase (r = -0.568, n = 20, P = 0.009). This suggested that within individual black carp, there is a trade-off between growth and maintenance (or development) of swimming performance under food-limited conditions.

Molecular characterization of Beclin 1 in rare minnow (Gobiocypris rarus) and its expression after waterborne cadmium exposure.

Beclin 1 plays an important role in autophagy and apoptosis which are well documented in mammals. However, relevant reports are rare in fish. This study characterized Beclin 1 of the rare minnow Gobiocypris rarus (rmBeclin 1), which encodes a peptide of 447 amino acids using RT-PCR and RACE. The deduced peptide showed 96.4 and 80.8% similarity to Beclin 1 of common carp and human, respectively. Semiquantitative RT-PCR revealed that rmBeclin 1 was ubiquitously expressed in all tested tissues of male and female fish in all developmental stages, even unfertilized eggs. RT-qPCR revealed that rmBeclin 1 mRNA transcripts were significantly up-regulated in gills after a 12 h treatment with waterborne CdCl2 but were decreased thereafter. However, rmBeclin 1 expression was decreased in the brain, but it was not significantly changed in other tissues. Subchronic CdCl2 exposure significantly increased rmBeclin 1 in the brain, but it distinctly decreased rmBeclin 1 in the gill and hepatopancreas. A dose-dependent effect was not observed in mature fish treated for 96 h, but a dose-dependent effect existed in immature fish treated for 10 days. Longer treatment (10 day) caused a significantly higher expression of rmBeclin 1 in the larvae groups. These data suggest that alterations in rmBeclin 1 after CdCl2 exposure are tissue-specific and time-related and that the dose-dependent effect was restricted to a certain concentration range and exposure time.

The effect of temperature on repeat swimming performance in juvenile qingbo (Spinibarbus sinensis).

To investigate the effect of temperature on the repeat constant acceleration swimming performance and on the metabolic recovery capacity in juvenile qingbo (Spinibarbus sinensis), their constant acceleration test speed (U(CAT)) and excess post-exercise oxygen consumption (EPOC) recovery process were measured twice with 1-h intervals at different acclimation temperatures (10, 15, 20, 25 and 30 °C). Temperature significantly affected U(CAT), the pre-exercise metabolic rate (MO(2)), metabolic peak values (MO(2peak)), the metabolic scope (MS, MO(2peak)--pre-exercise MO(2)) and the magnitude of the EPOC (P < 0.05). These parameters significantly increased as the temperature increased from 15 to 25 °C and significantly decreased (U(CAT) and EPOC magnitude) or did not change (MO(2peak) and MS) when the temperature increased from 25 to 30 °C in the first test (P < 0.05). The relationships between temperature (T) and these parameters (U(CAT), MO(2peak), MS and EPOC magnitude) in the first test were as follows: U(CAT) = 62.14/{1 + [(T - 25.1)/21.1](2)} (r = 0.847, P < 0.001, n = 40); MO(2peak) = 1,052.11/{1 + [(T - 29.2)/18.9](2)} (r = 0.901, P < 0.001, n = 39); MS = 753.74/{1 + [(T - 27.1)/18.6](2)} (r = 0.768, P < 0.001, n = 39); and EPOC = 195.42/{1 + [(T - 25.6)/8.7](2)} (r = 0.752, P < 0.001, n = 39). The optimal temperatures for U(CAT), MO(2peak), MS and EPOC magnitude in juvenile qingbo were 25.1, 29.2, 27.1 and 28.6 °C, respectively. Repeat exercise had different effect on U(CAT) and EPOC magnitude at different temperature (interaction effect, P < 0.05). There was no difference in U(CAT) and in EPOC magnitude between the first and second tests at low temperatures (10-20 °C). However, both U(CAT) and EPOC magnitude decreased significantly during the second test compared with the first test at high temperatures (25 and 30 °C) (P < 0.05). The present study showed that the recovery of the constant acceleration swimming performance was poorer at higher temperatures than at low temperatures in juvenile qingbo. These differences may be related to larger anaerobic metabolism, a lower pH value in the blood, larger ionic fluids and/or higher levels of hormones present at high temperatures.

[Application of Semi-solid Culture and Rapid Screening Strategy in the Preparation of Monoclonal Antibody against Plasmodium falciparum Histidine-rich Protein].

OBJECTIVE: To prepare the monoclonal antibody of Plasmodium falciparum histidine-rich protein, and analyze the roles of semi-solid culture technique and screening strategies in the preparation of monoclonal antibodies. METHODS: BALB/c mice was immunized with the recombinant antigen of Plasmodium falciparum histidine-rich protein (rePf-HRP). The spleen cells of immunized mice were fused with SP20 cells. The fused cells were cultured in semi-solid medium containing 2% methylcellulose to promote colony growth. The single cell clone was transferred to a liquid medium. Testing methods for screening culture supernatant was established based on the immune antigen and other related antigens. The positive cell clones by coarse screening and specific screening were preserved in liquid nitrogen. The positive cell lines were used for ascite antibody preparation, identification of IgG subclass, recognition sites, antibody affinity and application analysis on sensitivity of detecting antigen. RESULTS: A total of 915 cell clones were obtained in semi-solid culture after cell fusion. The positive rate by coarse screening was 37.8% (346/915). The positive rate of specific screening accounted for only 2.6% (9/346) of the coarse screening-positive clones, 0.98% (9/915) of the total number of clones. Eight specific antibody-secreting cell clones were obtained after liquid nitrogen frozen recovery tests. After further detection, 2 specific cell clones could be used as a pair of antibody for rapid detecting circulating antigen in the blood of patients with falciparum malaria. CONCLUSION: Semi-solid culture method can provide enough fused cells for screening. Combined strategy of coarse and specific screening ensures the rapid selection of specific clones.

Complete feminization of catfish by feeding Limnodilus, an annelid worm collected in contaminated streams.

BACKGROUND: Feminization of animals derived from areas polluted by endocrine disrupting chemicals (EDCs) has been observed in all classes of vertebrates. However, feminization of artificially reared offspring by feeding of specific living organisms has never been reported. METHODS: Different food (including Limnodilus spp collected from the wild) and time treatment were applied to southern catfish. In addition, EDCs in Limnodilus spp., an annelid worm collected from wild contaminated small streams, was detected by LC-MS (Liquid chromatography-mass spectrometry). Serum estradiol-17ß and vitellogenin (VTG) levels and gonadal Sf1, Dmrt1, Foxl2, Cyp19a1a expression levels in the catfish were measured through Estradiol/VTG EIA Kit and real-time PCR. RESULTS: Here we report that feeding of Limnodilus spp. resulted in complete feminization of southern catfish, which has a 1:1 sex ratio in wild conditions. Furthermore, HPLC analysis showed that the extraction of Limnodilus spp. contained EDCs, including bisphenol A (BPA), diethylstilbestrol (DES), 4-tert-octylphenol (4-t-OP) and 4-nonylphenol (4-NP), which were further confirmed by LC-MS. Feeding southern catfish using commercial diets sprayed with EDCs cocktail also resulted in 100% female, whereas the control fish displayed approximate 1:1 sex ratio. Limnodilus spp. fed fish displayed similar serum estradiol-17ß and VTG levels and gonadal Sf1, Dmrt1, Foxl2, Cyp19a1a expression levels to those of female control. CONCLUSION: These results demonstrated that EDCs in Limnodilus spp. cause southern catfish feminization by affecting aromatase expression and endogenous estrogen level. This is the first report showing that feeding of any living organism resulted in complete feminization of a vertebrate.

[Correlation of androgen receptor CAG repeats with the risks of benign prostatic hyperplasia and prostate cancer: a meta-analysis].

OBJECTIVE: To explore the association of the androgenic receptor (AR) CAG repeats with the risks of benign prostatic hyperplasia (BPH) and prostate cancer (PCa). METHODS: We searched the major databases at home and abroad for the literature addressing the correlation of the AR gene CAG repeats with BPH and PCa. Based on the results of heterogeneity tests, we used the M-H fixed effect model and random effect model to pool the odds ratio (OR) effect size. We evaluated publication bias by Begg and Egger bias analysis, investigated the association of CAG repeats with the risks of BPH and PCa by systematic review, and stratified their relationship according to the races of the patients. RESULTS: Based on the selection criteria, 4 of the 29 identified studies were included, with 485 cases of BPH, 767 cases of PCa, and 709 controls. There was no heterogeneity between the BPH and control groups, and no correlation between short CAG repeats and BPH after pooling the odds ratio (OR) effect size. Heterogeneity was found among the BPH, PCa and control groups. Random effects model suggested an association of short CAG repeats with the risk of PCa (OR(PCa/control) = 1.45, OR(PCa/BPH) = 1.86, OR(PCa/(BPH + control)) = 1.66), while subgroup analysis with racial stratification indicated inter-ethnic differences between the two. Begg and Egger bias analysis showed no significant publication bias. CONCLUSION: Shorter CAG repeats are positively correlated with the risk of PCa but not with that of BPH.

[Comparison of two rapid diagnostic tests in detection of malaria parasites].

OBJECTIVE: To compare the performance of two rapid diagnostic tests (RDTs) for malaria parasite detection. METHODS: Blood samples of 200 malaria patients and 60 non-malaria persons were collected from Yunnan and Shanghai, respectively. The sera were detected by gold-colloidal immunochromatography (GICA) and OptiMAL, and microscopy was used as gold standard in species identification. The sensitivity, specificity, minimum detection limit of the two RDTs was compared. RESULTS: Of the 260 samples, malaria parasites were found in 200 by microscopy, of which 100 each were Plasmodium falciparum and P. vivax, respectively. Compared with microscopy, the sensitivity and specificity of GICA and OptiMAL for the samples were 87.5% (175/200) and 93.3% (56/60), 95.5% (191/200) and 100.0% (60/60), respectively. The sensitivity and specificity of GICA and OptiMAL for detection of P. falciparum were 83.0% (83/100) and 96.9% (155/160), 90.0% (90/100) and 99.4% (159/160), respectively; and for detection of P. vivax, they were 89.0% (89/100) and 98.8% (158/160), 96.0% (96/100) and 97.5% (156/160), respectively. There was a significant difference in malaria detection between GICA and OptiMAL (chi2 = 8.23, P < 0.05). No statistical difference was found between the two RDTs in P. falciparum and P. vivax detection (P > 0.05). OptiMAL showed better result in detection of P. falciparum when the parasite density was higher. The minimum detection limit of the two RDTs was about 100-200 parasites/microl blood. CONCLUSION: Compared to GICA, OptiMAL has higher sensitivity and specificity. However, GICA shows lower minimum detection limit and better reproducibility in blood samples with different densities than that of OptiMAL.

The effect of aerobic exercise training on growth performance, digestive enzyme activities and postprandial metabolic response in juvenile qingbo (Spinibarbus sinensis).

Continual swimming exercise usually promotes growth in fish at a moderate water velocity. We hypothesized that the improvement in growth in exercise-trained fish may be accompanied by increases in digestive enzyme activity, respiratory capacity and, hence, postprandial metabolism. Juvenile qingbo fish (Spinibarbus sinensis) were subjected to aerobic training for 8weeks at a water velocity of control (3cms(-1)), 1, 2 and 4 body length (bl)s(-1) at a constant temperature of 25°C. The feed intake (FI), food conversion rate (FCR), specific growth rate (SGR), whole-body composition, trypsin and lipase activities, maximal oxygen consumption (M˙O2max) and postprandial M˙O2 response were measured at the end of the training period. Aerobic exercise training induced a significant increase in FI compared with the control group, while the FCR of the 4bls(-1) group was significantly lower than for the other three groups (P<0.05). The 1 and 2bls(-1) groups showed a significantly higher SGR over the control group (P<0.05). The whole-body fat and protein contents were significantly altered after aerobic exercise training (P<0.05). Furthermore, aerobic exercise training elevated the activity of both trypsin and lipase in the hepatopancreas and intestinal tract of juvenile S. sinensis. The M˙O2max of the 4bls(-1) training group was significantly higher than for the control group. The resting M˙O2 (M˙O2rest) and peak postprandial M˙O2 (M˙O2peak) in the three training groups were significantly higher than in the control group (P<0.05). Time to M˙O2peak was significantly shorter in the 1, 2 and 4bls(-1) training groups compared with the control group, while exercise training showed no effect on SDA (specific dynamic action) duration, factorial metabolic scope, energy expended on SDA and the SDA coefficient when compared to the control group. These data suggest that (1) the optimum water velocity for the growth of juvenile S. sinensis occurred at approximately 2.4bls(-1); (2) the improvement of growth may have been primarily due to an increase in the FI after long-term training; (3) and aerobic exercise training boosted the activity of digestive enzymes and maximum digestive metabolism, which could favor fast digestion and growth in juvenile S. sinensis.

[Association of TET2, LMTK2 and FAM84B gene expression with prostate cancer risk in Chinese patients].

OBJECTIVE: To explore the association between the common variations of TET2 (rs7679673, A), MTK2 (rs6465657, T) and FAM84B (rs12543663, C) genes and prostate cancer (Pca) risk in Chinese population in Beijing, and to understand the relationship between genotypes and phenotypes including clinical characteristics and life style, etc. in patients with prostate cancer. METHODS: Based on a case-control study, 124 patients with prostate cancer and 138 age-matched control subjects were recruited. Information of clinical phenotype and life style, etc. in the prostate cancer patients was collected. We compared the differences of allele and genotype frequencies of TET2 (rs7679673, A), LMTK2 (rs6465657, T) and FAM84B (rs12543663, C) gene expressions between the two groups for the allele and genotype frequencies, and explored the relationship between different genotypes and clinical features such as patient age, BMI, Gleason score, PSA level and tumor stage, by Chi-square test in patients with PCa. Multifactor dimensionality reduction was used to detect the gene-gene interactions. RESULTS: The FAM84B (rs12543663, C) C carriers frequency had significant difference between the case group and the control group (χ(2) = 3.980 P = 0.046; OR = 1.883; 95%CI = 1.006-3.526). The allele and genotype frequencies of TET2 gene (rs7679673, A) and LMTK2 gene (rs6465657, T) were not significantly different between the case group and the control group (P > 0.05). Analysis of the genotypes and clinical phenotypes showed that the genetic type of FAM84B C carriers [CX (CC + CA)] were significantly associated with cancer stage (χ(2) = 9.585; P = 0.002; OR = 3.740; 95%CI = 1.580 - 8.853). Association between three loci and 12 kind of relevant outcomes was found in TET2 A carriers and the smoking and drinking patients (all P < 0.05). Significant correlation was also found between LMTK2 (rs6465657, T) TX carriers and surgery (χ(2) = 8.612; P = 0.003; OR = 0.174; 95%CI 0.049 - 0.620). No significant correlation was seen with other covariates (P > 0.05). Dendrogram analysis among the three loci showed that the best model consisted of the three sites (P = 0.0270), cross validation consistency: 10/10, and testing balanced accuracy: 0.5120. There may be gene-gene interaction among TET2 (rs7679673, A), LMTK2 (rs6465657, T), and FAM84B (rs12543663, C). CONCLUSIONS: There may be obvious association of FAM84B (rs12543663, C) gene with prostate cancer risk and the stages, and the synergistic effects of TET2 (rs7679673, A), LMTK2 (rs6465657, T) and FAM84B (rs12543663, C) genes may have an association with prostate cancer risk in Chinese population.

Synonymy and misidentification of three Hahnia species (Araneae: Hahniidae) from China.

The type specimens of two Hahnia species, H. maginii Brignoli, 1977 and H. thorntoni Brignoli, 1982 were examined to determine the identification of Hahniidae from South China. Hahnia thorntoni is found to be a senior synonym of H. flagellifera Zhu, Chen & Sha, 1989, while the paratype male of H. thorntoni belongs to another species, H. zhejiangensis Song & Zheng, 1982. Chinese specimens previously identified as H. maginii probably belong to H. thorntoni. The female and male specimens of H. yueluensis Yin & Wang, 1983 were mismatched and misidentified; the female holotype and paratype belong to H. thorntoni and male allotype to H. zhejiangensis.

[Serodiagnosis of the recombinant multi-epitope antigens from antigen B subunits of Echinococcus granulosus].

OBJECTIVE: To evaluate the diagnostic value of six recombinant multi-epitope antigens and three AgB subunit antigens from antigen B subunit of Echinococcus granulosus. METHODS: A linker sequence was inserted into the sequence of MEA-26 to make it an MEA-49. I-TASSER on-line server was used to analyze protein structure. The reactivity of two multi-epitope recombinant antigens with the same target sequence but in different tertiary structure was compared. The reactivity of six multi-epitope antigens (MEA-8, MEA-20, MEA-26, MEA-36, MEA-49, and MEA-52), 3 subunit antigens (AgB1, AgB2, and AgB4), and 2 control antigens (Trx and linker) was determined by indirect ELISA. The assays were performed on 232 serum samples separated as follows: 112 sera from patients with cystic echinococcosis, 35 sera from individuals with alveolar echinococcosis, 43 sera from patients with cysticercosis and 42 sera from healthy individuals. Their diagnostic performance was assessed by receiver operating characteristic (ROC) curve analysis. RESULTS: Tertiary structure prediction showed that the epitope regions of MEA-26 were closer to each other and aligned in parallel, while that in MEA-49 were farther apart from each other and formed two independent domains. Serological analysis revealed that the mean P/N value (2.88 +/- 2.02), sensitivity (92%) and diagnostic efficiency (89%) of MEA49 were higher than that of MEA-26 (2.54 +/- 2.02, 78% and 82%). MEA-20 (2.24 +/- 1.31), MEA-26 (2.54 +/- 2.02), MEA-36 (2.44 +/- 1.51), MEA-49 (2.88 +/- 2.02) and MEA-52 (2.50 +/- 1.37) showed a high reactivity to the sera from patients with cystic echinococcosis, which was superior to that of AgB1 (2.15 +/- 1.26). There was no significant difference in the reactivity to sera from individuals with alveolar echinococcosis between multi-epitope antigens and AgB1 (P > 0.05). MEA-52 showed a high diagnostic sensitivity in cysticercosis cases (1.27 +/- 0.70), superior to that of AgB1 (0.95 +/- 0.13) (P < 0.01). The reactivity of MEA-8 (1.04 +/- 0.15) to sera from healthy individuals was significantly higher than that of AgB1 (0.89 +/- 0.07) (P < 0.01). ROC analysis showed in the cases of cystic echinococcosis, the diagnostic sensitivity accomplished with AgB1, AgB2, and AgB4 was 77%, 55%, and 66%, respectively; the multi-epitope antigens of MEA-49 (92%), MEA-36 (92%), MEA-52 (87%), and MEA-26(78%) revealed a higher sensitivity than AgB1. CONCLUSION: The reactivity of multi-epitope antigens is superior to that of AgB subunit antigens. The reactivity of MEA-49 is higher than that of MEA-26 which has the same target sequence but in different tertiary structure.

Exposure to acute waterborne cadmium caused severe damage on lipid metabolism of freshwater fish, revealed by nuclear lipid droplet deposition in hepatocytes of rare minnow.

Cadmium (Cd) is a widely distributed aquatic toxic heavy metal with the potential to disrupt fish metabolism; however, more research is needed to clarify the underlying mechanisms. In the present study, rare minnows (Gobiocypris rarus) were used to detect the effects of cadmium on freshwater fish lipid metabolism and its underlying mechanism by histopathological observation, measurement of serum and liver biochemical indexes, and analysis of gene expression in terms of lipid oxidation, synthesis and transport. Here, severe damage, such as cytoplasmic lipid droplet (LD) accumulation, ectopic deposition of LDs, and the appearance of nuclear LDs (nLDs), was detected after exposure to 2.0 mg/L or higher concentrations (2.5 and 2.8 mg/L CdCl2) for 96 h. Other damage included abnormal increases in rough endoplasmic reticulum (RER) lamellae in a fingerprint or concentric circle pattern and necrosis of hepatocytes, and which was observed in the livers of fish exposed to 2.0 mg/L CdCl2.. Both hepatic and serum lipids, such as triglycerides and total cholesterol, were significantly increased after exposure to 2.0 mg/L CdCl2, as was serum lipase (LPS). Hepatic lipase and lipoprotein lipase remained unchanged, in accordance with the unchanged hepatic mRNA transcripts of PPARɑ. Furthermore, the mRNA transcripts of both SCD and SQLE were significantly decreased. Moreover, hepatic and serum low-density and high-density lipoprotein cholesterol showed significant changes, which were accompanied by a significant increase and decrease in hepatic APOAI and APOB100 mRNA levels, respectively. All the results indicate the presence of severe damage to hepatic lipid metabolism and that disrupted lipid transport may play a key role in the accumulation of hepatic LDs. In addition, the hepatic nLDs of nonmammalian vertebrates and their location across the nuclear envelope are intriguing, suggesting that large-size nLDs are a common marker for severe liver damage.

Application of the first rechargeable sacral neuromodulation system for treatment of neurogenic lower urinary tract dysfunction in China: a case report.

Neurogenic lower urinary tract dysfunction (NLUTD) is caused by nervous system lesions and characterized by impaired micturition and urinary incontinence. The goal of treatment is to manage these symptoms, improve quality of life, prevent urinary tract infections, and maintain urinary function. Pelvic floor muscle training and medication are commonly used for treating it. Sacral neuromodulation (SNM) has been used in the treatment of NLUTD for >20 years worldwide, and its effectiveness and safety have been verified. Several countries have begun using a rechargeable SNM system, whereas the current sacral SNM system used in China is non-rechargeable. A 29-year-old man with persistent voiding dysfunction for >20 years presented with progressive symptoms 1 year ago. He was admitted to our hospital in August 2022 for a rechargeable SNM system implantation. The patient underwent a video-urodynamic examination and the Short Form of a Urinary Quality of Life Questionnaire (SF-Qualiveen) before and 1 month after surgery. The video-urodynamic examination showed that the maximum bladder capacity significantly increased after surgery, bladder compliance improved, the phenomenon of uninhibited bladder contraction during filling decreased, and urine leakage was reduced. The SF-Qualiveen score showed the patient's quality of life significantly improved. To our knowledge, this is the first case of a rechargeable SNM system implantation in China, which shows that it is safe and effective. More clinical cases and long-term observation are still needed. In conclusion, a rechargeable SNM system has significance for health and the economy and has a broad clinical application prospect.

Analysis on the reactivity of five subunits of antigen B family in serodiagnosis of echinococcosis.

In this study, the reactivity and differences of five subunits of echinococcus antigen B (AgB) family, recognizing specific antibodies in echinococcosis patient serum, were analyzed. Eight recombinant subunit antigens from Echinococcus granulosus (EgAgB1-EgAgB4) and Echinococcus multilocularis (EmAgB1-EmAgB3 and EmAgB5) were tested by ELISA using a panel of 243 serum samples collected from cystic echinococcosis (CE), alveolar echinococcosis (AE), cysticercosis (CC) patients and clinically normal individuals (NH). The results showed that the diagnostic sensitivity of the subunits for CE sera were 83.06%, 62.90%, 29.03%, 75.81% and 41.13%, and the specificities were 73.95%, 72.27%, 76.47%, 73.11% and 85.71%, respectively. The reactivity of three paralogous subunits, EgAgB1, EgAgB2 and EgAgB3 from E. granulosus and EmAgB1, EmAgB2 and EmAgB3 from E. multilocularis were compared by serological assay. All of the orthologous subunits showed no statistical difference (P>0.05) in detecting CE and AE sera; it revealed that the reactive epitopes may be similar between the orthologous subunits. In a total of 124 CE sera, the positive recognition rate by EgAgB1 was the highest (103/124), yet cocktail subunit antigens may detect even more positives from 100/124 to 112/124 using different subunit combinations. IgG4 subclass was the predominant antibody in reacting with subunit antigens. To conclude, the epitopes of orthologous AgB subunits from E. granulosus and E. multilocularis that recognize specific antibodies may be similar. The paralogous subunits EgAgB1, EgAgB2 and EgAgB4 were the main reactive subunit in sera detection and may have utility as echinococcosis diagnostics, with EgAgB1 possessing the greatest potential. Cocktail subunits may improve the positive detection rate.