[Efficacy and safety of low-dose rasburicase for refractory chronic gouty arthritis].

Objective: To explore the efficacy and safety of low-dose rasburicase for refractory chronic gouty arthritis. Methods: A cohort study. The clinical data of patients with refractory chronic gouty arthritis who were treated with rasburicase at Sun Yat-sen Memorial Hospital, Sun Yat-sen University between January 2021 and July 2022 were retrospectively analyzed. Refractory chronic gouty arthritis was defined as serum uric acid (sUA)>360 µmol/L and urate volume>10 cm3 under dual-energy computed tomography after tolerable maximal oral urate-lowering therapy for at least 3 months. The administration of low-dose rasburicase was applied intravenously with total dosage ranging from 4.5 to 7.5 mg each dose, at 4-week intervals for a maximum of three doses. Efficacy was evaluated by the changes of sUA level, tophus and urate volume. Results: A total of 22 patients were included for analysis, with 95.4% (21/22) male, the mean age was (44±15) years, and the median duration of gout was 11 (6-15) years. The mean sUA at baseline was (667±112) µmol/L. The levels of sUA significantly decreased after each dose of rasburicase (P<0.001), and the median reduction of sUA after each dose of rasburicase was 568 (471-635), 187 (66-335) and 123 (49-207) µmol/L, respectively. At week 12, nine patients (40.9%) exhibited sUA<360 µmol/L and tophus disappeared in one patient. The urate volume significantly decreased at week 12 when compared with that before the first dose of rasburicase in all the patients [40 (16-172) cm3 vs 17 (7-134) cm3, P<0.001], with a median reduction rate of 41.6% (22.9%-58.5%). The everall safety of rasburicase was good, and no serious adverse reactions occurred. Conclusions: Low-dose rasburicase is well-tolerated and effective for decreasing the urate burden in patients with refractory chronic gouty arthritis. Further prospective randomized controlled trials are needed to validate these findings.

[Predictive value of anti-mutated citrullinated vimentin antibody on one-year radiographic progression in patients with rheumatoid arthritis].

Objective: To investigate the value of baseline anti-mutated citrullinated vimentin (MCV) antibody for predicting one-year radiographic progression in patients with rheumatoid arthritis (RA). Methods: Consecutive RA patients were recruited from November 2014 to July 2018 at Department of Rheumatology, Sun Yat-sen Memorial Hospital, Clinical data were collected including disease activity score in 28 joints with four variables including C-reactive protein (CRP).Serum anti-MCV antibody at baseline was detected by enzyme-linked immunosorbent assay. X ray assessment of both hands/wrists was performed and assessed according to the Sharp/van der Heijde modified score (mTSS) at baseline and the 12th month. Univariate and multivariate logistic regression analyses were used to identify the risk factors for one-year radiographic progression. Results: Among 220 RA patients recruited, the positive rate of anti-MCV antibody at baseline was 77.7%. Compared with those with negative anti-MCV antibody, RA patients with positive anti-MCV antibody had higher disease activity score in 28 joints with four variables induding CRP [3.8 (2.4, 5.0) vs. 3.1 (2.1, 4.0), P=0.007], more physical dysfunction (21.6% vs. 8.2%, P=0.033) and higher radiographic indicators including mTSS [11 (2, 27) vs. 4 (1, 10), P=0.003], joint space narrowing [JSN, 4 (0, 14) vs. 2 (0, 6), P=0.024] and joint erosion[JE, 5 (1, 18)vs. 3 (0, 5), P=0.003]. After one-year follow-up, sixty-six RA patients (30.0%) developed radiographic progression, the percentage of whom was significantly higher in positive anti-MCV group than that in negative anti-MCV group (33.9% vs.16.3%, P=0.018). Multivariate logistic regression analysis suggested that positive anti-MCV antibody at baseline was an independent risk factor for one-year radiographic progression (OR=2.341, 95%CI 1.002-5.469). Conclusion: Positive anti-MCV antibody at baseline predicts one-year radiographic progression in RA patients. In the future, anti-MCV antibody can be used not only as a supplementary laboratory marker, but also in disease activity assessment and prognosis prediction for RA.

[The mechanism of circ_0023990/miR-873-5p/ANXA2 axis regulating radiosensitivity and development of thyroid carcinoma].

Objective: To explore the effect and possible mechanism of circ_0023990 on the radiosensitivity of thyroid cancer cells. Methods: qRT-PCR was used to detect the expression of circ_0023990 in the cancer tissues of 55 patients with thyroid cancer and thyroid cancer cell lines (TPC-1, KTC-1, FTC-133 and CAL-62), and the relationship between the expression of circ_0023990 in cancer tissues and the clinical characteristics of the patients were analyzed. Thyroid cancer cells TPC-1 and KTC-1 were divided into sh-circ_0023990 group, sh-NC group, sh-circ_0023990+anti-miR-873-5p group, sh-circ_0023990+anti-miR-NC group, miR-873-5p group, miR-NC group, miR-873-5p+pcDNA-ANXA2 group and miR-873-5p+pcDNA group, and then clone formation experiment was used to detect cell radiosensitivity. After each group of cells was irradiated with 4Gy radiation, the expression of γH2AX protein in the cells was detected by Western Blot. The dual luciferase reporter gene experiment verified the targeting relationship between circ_0023990 and miR-873-5p or miR-873-5p and ANXA2. Results: The expression of circ_0023990 in thyroid cancer tissues was higher than that in normal tissues (2.15±0.09 vs. 0.97±0.05, P<0.05), and its expression was closely related to tumor size, lymph node metastasis and TNM staging of patients with thyroid cancer (P<0.05). The expression of circ_0023990 in thyroid cancer cell lines (TPC-1, KTC-1, FTC-133 and CAL-62) were higher than that of normal thyroid cells HTori-3 (3.16±0.38, 2.63±0.28, 1.82±0.24, 1.71±0.22 vs. 1.00±0.10, all P<0.05). The survival scores of TPC-1 and KTC-1 cells in the sh-circ_0023990 group were significantly lower than those in the sh-NC group (P<0.05), and the sensitization ratios were 2.482, 1.643; The survival scores of TPC-1 and KTC-1 cells in the sh-circ_0023990+anti-miR-873-5p group were higher than those in the sh-circ_0023990+anti-miR-NC group (P<0.05), and the sensitization ratios were 0.305, 0.441, respectively. The survival scores of TPC-1 and KTC-1 cells in the miR-873-5p group were lower than those in the miR-NC group (P<0.05), and the sensitization ratios were 2.044, 1.653 respectively. The survival scores of TPC-1 and KTC-1 cells in the miR-873-5p+pcDNA-ANXA2 group was higher than that in the miR-873-5p+pcDNA group (P<0.05), and the sensitization ratios were 0.496, 0.686, respectively. The expression of γH2AX protein in TPC-1 and KTC-1 cells of the 4 Gy+sh-circ_0023990 group were higher than that in the 4 Gy+sh-NC group (2.68±0.27 vs. 1.87±0.25, 2.46±0.19 vs. 1.77±0.14; all P<0.05), but the expression of γH2AX protein in TPC-1 and KTC-1 cells of the 4 Gy+sh-circ_0023990+anti-miR-873-5p group were lower than that in the 4 Gy+sh-circ_0023990+anti-miR-NC group (1.13±0.09 vs. 1.69±0.09, 1.11±0.08 vs. 1.60±0.08; both P<0.05). The expression of γH2AX protein in TPC-1 and KTC-1 cells in the 4 Gy+miR-873-5p group were higher than that in the 4 Gy+miR-NC group (2.35±0.16 vs. 1.84±0.14, 2.26±0.12 vs. 1.77±0.13; both P<0.05), but the expression of γH2AX protein in TPC-1 and KTC-1 cells of the 4 Gy+miR-873-5p+pcDNA-ANXA2 group were lower than that in the 4 Gy+miR-873-5p+pcDNA group (1.96±0.12 vs. 2.41±0.12, 1.92±0.07 vs. 2.28±0.12; both P<0.05). circ_0023990 targeted the negative regulation of miR-873-5p, and ANXA2 was the target gene of miR-873-5p. Conclusion: circ_0023990 was highly expressed in thyroid cancer tissues and cell lines, and it may promote the radiotherapy resistance of thyroid cancer cells in vivo through regulating miR-873-5p/ANXA2 axis.

Antimicrobial resistance and virulence genes of Streptococcus isolated from dairy cows with mastitis in China.

Streptococcus is a major mastitis-causing pathogen in dairy cows. To investigate the prevalence, antimicrobial resistance and virulence gene of Streptococcus in mastitic milk, a total of 735 mastitic raw milk samples from dairy cows in 11 provinces of China were collected and tested. Antimicrobial resistance of Streptococcus isolates was determined by disc diffusion against 8 classes 29 antimicrobial agents, and Streptococcus resistant genes and virulence genes were determined by PCR and agarose gel electrophoresis. A total of 64 (8.71%) isolates of Streptococcus were isolated and identified using biochemical profiling, including 22 isolates of Streptococcus agalactiae, 13 isolates of Streptococcus dysgalactiae, and 29 isolates of Streptococcus uberis. Out of 64 resistant Streptococcus isolates, all isolates (100%) were resistant to 3 or more antimicrobials. The most frequency (n = 18, 28.12%) of the isolates were multi-resistant to 5-7 antimicrobials and the highest multi-resistant number was 29 (n = 1, 1.56%). Streptococcus isolates had the highest resistance rate to tetracycline (98.44%) and oxacillin (98.44%), followed by penicillin G (96.88%) and doxycycline (96.88%), and the lowest resistance was observed with respect to ciprofloxacin (1.56%). A total of 16 antimicrobials resistance genes with 25 combination patterns were detected in the isolates. The gene combination of Sul1/Sul2/Sul3 + gyrA/parC + cat1/cat2 was the most common pattern (12.5%). The correlation between resistant phenotypes and resistance genes in Streptococcs was 35.87%. A total of 7 virulence genes were detected and 59 (92.19%) isolates harbored at least one gene. Twenty-four classes of gene patterns were found in the isolates and the patterns of bca (9.38%) and cfb (9.38%) were the most prevalent form. In conclusion, the issue of drug resistance of Streptococcus is still a great concern in cattle health in China.

Relative contributions of testosterone deficiency and metabolism syndrome at the risk of reduced quality of life: A cross-sectional study among Chinese mid-aged and elderly men.

Testosterone deficiency and metabolism syndrome (MetS) are universal among ageing males, and they have been suggested responsible for poorer quality of life (QoL). We aimed to evaluate the relative contributions of reproductive hormones and components of MetS at the risk of reduced QoL among Chinese mid-aged and elderly men. A cross-sectional study recruited 2,364 males aged 40-79 years, and 2,165 was included for analysis eventually. The Chinese version of ageing male symptoms scale, 36-item Short Form and Beck Depression Inventory were applied to assess QoL. Bivariate correlation analysis and multiple linear regression analysis were used to assess the relative contributions of reproductive hormones and components of MetS at the risk of reduced QoL. Testosterone deficiency and MetS contributed to poorer QoL, of which higher fasting blood glucose made the primary contribution, lower total testosterone mainly contributed to poorer physical functioning.

[Influence of high-fat diet in paternal C57BL/6 mice on liver fat deposition in offspring].

Objective: To investigate the influence of high-fat diet (HFD) in paternal C57BL/6 mice on HFD-induced liver fat deposition in male offspring, as well as transgenerational inheritance caused by paternal HFD and related mechanisms. Methods: A total of 20 male C57BL/6 mice aged 3 weeks (F0) were randomly divided into normal control group (C, 10 mice) and HFD group (HF, 10 mice). After 12 weeks of HFD intervention, the male mice in the HFD group mated with female ones treated with normal diet and pups were obtained. Male pups (F1) were selected as study subjects. According to the intervention for F0 mice, male F1 mice were divided into control male offspring group (CM, 8 mice) and HFD male offspring group (HFM, 9 mice). All these mice were given normal diet after weaning until 4 weeks old, followed by HFD for 4 weeks. The body length and body weight were measured and recorded every week. Oil red O staining was used to observe fat deposition in the liver. Western blot and real-time PCR were used to measure the expression of related proteins and genes involved in the de novo synthesis and aerobic oxidation of fatty acid, mitochondriogenesis, and autophagy. Results: After 4 weeks of HFD intervention, the HFM group had a significantly higher body weight than the CM group (P < 0.05); the oil red O staining showed that compared with the CM group, the HFM had a significant increase in liver fat deposition and a significantly higher integral absorbance value in the oil red O staining-positive area (384 360±57 600 vs 236 754±12 607, P < 0.01). For related factors involved in the de novo synthesis of fatty acid in the liver, compared with the CM group, the HFM group had significant increases in the expression of sterol regulatory element-binding protein-1 and fatty acid synthase (P < 0.05); for related factors involved in the mitochondrial biosynthesis in the liver, the HFM group had significant reductions in the relative expression of peroxisome proliferator-activated receptor-γ coactivator-1α, nuclear respiratory factor 1, and mitochondrial transcription factor A compared with the CM group (P < 0.05). For autophagy-related factors in the liver in the F1 mice, compared with the CM group, the HFM group had a significant reduction in microtubule-associated protein I light chain 3 (LC3-II/I) (P < 0.05) and a significant increase in P62 (P < 0.05), suggesting a reduced autophagy function in the liver. Conclusion: HFD intervention for paternal C57BL/6 mice can increase HFD-induced liver fat deposition in male offspring, which may be related to the increased de novo synthesis of fatty acid and reduced mitochondriogenesis and autophagy function in the liver.

Importance of CD44 on umbilical cord mesenchymal stem cells for expansion of hematopoietic cells.

Human umbilical cord mesenchymal stem cells (hUCMSCs) have important functions on the expansion of hematopoietic stem cells (HSCs) through providing the essential microenvironment for hematopoiesis. In order to test whether CD44 on hUCMSCs could have a key function for the ability of hUCMSCs to expand human HSCs, the soluble anti—CD44 antibody was added to the co—cultures of hUCMSCs and cord blood (CB) CD34+ cells, which blocked the ability of hUCMSCs to expand CB CD34+ cells significantly. Long—term culture initiating cell (LTC—IC) assay revealed that the ability of multipotent differentiation of CB CD34+ cells co—cultured with CD44 knockdown hUCMSCs could only retain lasting at most for 5 weeks in vitro. In vivo assay, based on non—obese diabetic/severe combined immunodeficient disease (NOD/SCID) mice, revealed that the hematopoietic reconstitution potential of CB CD34+ cells co—cultured with CD44 knockdown hUCMSCs is significantly reduced. The hematopoietic supporting ability of hUCMSCs in vivo and in vitro is reduced upon the knockdown of CD44. CD44 has important functions on the ability of hUCMSCs to expand human HSCs in the cell— extrinsic control.

IL-8 -251T/A polymorphism is associated with susceptibility to acute pancreatitis.

We conducted a case-control study to clarify the asso-ciations between inflammatory cytokine, including interleukin (IL)-1b, IL-6, IL-8, and IL-10, polymorphisms and risk of acute pancreatitis. Genotyping analyses of IL-1ß+3954 C/T (rs1143634), IL-1ß-511 C/T (rs16944), IL-6 -174 G/C (rs1800795), IL-6 -634 C/G (rs1800796), IL-8 -251T/A (rs4073), IL-10 -1082A/G (rs1800896), and IL-10 -819C/T (rs1800871) were conducted using polymerase chain reaction-restriction fragment length of polymorphism. Unconditional logistic regression analysis was utilized to assess the potential association be-tween genotype frequencies and risk of acute pancreatitis. Multivari-ate regression analyses showed that subjects carrying the IL-8 -251 AA genotype had a significantly increased risk of acute pancreatitis, with an adjusted odds ratio (95% confidence interval) of 1.55 (1.02-2.36). However, we found no significant association between IL-1ß +3954 C/T, IL-1ß -511 C/T, IL-6 -174 G/C, IL-6 -174 G/C, IL-6 -634 C/G, IL-10 -1082A/G, or IL-10 -819C/T polymorphisms and risk of acute pancreatitis. We found that the IL-8 -251T/A polymorphism was associated with a higher susceptibility to acute pancreatitis in a Chinese population.

Development and characterization of SSR markers from Pinus massoniana and their transferability to P. elliottii, P. caribaea and P. yunnanensis.

Pinus massoniana (Masson's pine) is a widespread tree species in central and southern China and northern Vietnam; it is valued for rosin and paper production. Despite the significant economic value of Masson's pine, little work has been done on its molecular genetics. We developed 318 SSR primers from genome sequences of P. massoniana, and we identified 10 polymorphic markers. The number of alleles in the population of P. massoniana that we examined ranged from two to four, and the Shannon diversity index ranged from 0.150 to 1.133. Cross-species transferability of the 318 SSRs was also analyzed in the slash pine (Pinus elliottii), the Caribbean pine (Pinus caribaea) and the Yunnan pine (Pinus yunnanensis); 15, 10, and 10 primer pairs generated polymorphic amplification, respectively. These sets of polymorphic SSR markers will be useful for population genetics studies of P. massoniana, for genetic identification of interspecific hybridization, and for phylogeographic studies of Pinus spp.

Effects of heat stress on survival of Frankliniella occidentalis (Thysanoptera: Thripidae) and Thrips tabaci (Thysanoptera: Thripidae).

Temperature is known to play a crucial role in the population dynamics of insects. Insects have evolved different mechanisms to resist unfavorable extreme temperatures. In recent years, western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), and onion thrips, Thrips tabaci (Lindeman) (Thysanoptera: Thripidae), have caused significant damage to vegetable crops. Because of global warming and expanding areas of vegetable cultivation, a study of the effects of heat stress on these thrips species is warranted. We exposed the various developmental stages of western flower thrips and onion thrips to temperatures of 41, 43, or 45 degrees C for 2, 6, 12, 24, or 36 h to determine the effects of heat stress on survival. Our results showed that the heat resistance of nonadult western flower thrips was greater than that of the nonadult onion thrips, and that the natural heat resistant ability was the primary factor in heat resistance in western flower thrips. In contrast, the heat resistance of adult onion thrips was greater than that of the adult western flower thrips, which was primarily the result of the ability of searching suitable microenvironment that enabled the onion thrips to mitigate the effects of high temperatures more efficiently than the western flower thrips. Our analysis of the differences in heat resistance between western flower thrips and onion thrips provides important information for the development of thermal treatments for controlling western flower thrips and onion thrips.

Cystic fibrosis transmembrane conductance regulator protein expression rate in healthy spermatozoa is not correlated with ovum fertilisation rate.

Our previous studies have shown that the cystic fibrosis transmembrane conductance regulator (CFTR) was important for capacitation and fertilisation in mouse, guinea pig and human spermatozoa. However, it is unclear whether CFTR is correlated with ovum fertilisation rate. The present study was to test the possible relationship between spermatozoa CFTR protein expression rate in healthy men and ovum fertilisation rate during in vitro fertilisation. Ninety-four couples for female factor infertility for IVF-ET treatments were retrospectively studied. All the patients were divided into three groups based on the fertilisation rate of ovum in vitro. It was performed to explore whether there were differences in sperm CFTR protein expression rate among the three groups and the relevance between CFTR protein expression rate and ovum fertilisation rate. Our study showed that there was no significant differences in sperm CFTR protein expression rate among the three groups (F = 0.614, P = 0.544), and the relevance between spermatozoa CFTR protein expression rate and ovum fertilisation rate was not significantly different (r = 0.013, P = 0.904). These results further suggest that CFTR protein expression rate in healthy men spermatozoa was not associated with ovum fertilisation rate and thus we cannot predict ovum fertilisation results by sperm CFTR protein expression rate.

[Targeted therapy of pyogenic sterile arthritis, pyoderma gangrenosum, and acne syndrome (PAPA): a case report and literature review].

Objective: To analyze the clinical course and targeted therapy of pyogenic sterile arthritis, pyoderma gangrenosum, and acne (PAPA) syndrome. Methods: The clinical history of a 6-year-old boy with PAPA syndrome, who was admitted to Hong Kong University Shenzhen Hospital in September 2017, was reviewed. His genetic diagnosis was confirmed by whole exome sequencing. The response to targeted therapy was evaluated by comparing the inflammatory markers (erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) and serum cytokines (interleukin (IL)-1, IL-6 and tumor necrosis factor-α (TNF-α)) before and after biological agents treatment. For literature review, "PAPA syndrome" and"PSTPIP1 gene"were used as keywords to retrieve papers published from January 1997 to December 2019 from Pubmed, Wanfang and CNKI database. Results: The patient was a 6-year-old boy, admitted to the hospital due to recurrent joint swelling and pain for more than 4 years. Before treatment, the CRP (256 mg/L), ESR (105 mm/1 h) and cytokines including serum TNF-α (7.43 ng/L), IL-1 (<5 ng/L), IL-6 (301 ng/L) were significantly elevated. Culture of the joint effusion was negative, but the IL-6 level was above 1 000 ng/L. MRI showed osteomyelitis at the lower end of the right femur. Gene detection found a heterozygous variation of PSTPIP1 gene (c.748G>A, p.E250K). Arthralgia once alleviated after the initiation of tocilizumab and infliximab, but recurred after 1 year of treatment. Thereafter, the anti-IL-1 receptor antagonist (Anakinra) was commenced, followed by a significant improvement of the arthralgia, and a complete remission during the follow-up. Besides, the level of CRP, ESR, serum TNF-α, IL-1 and IL-6 were all decreased to normal on the last followed up in December 2019. Literature review found 29 articles and 87 patients in total. The initial symptoms included those of arthritis (n=58), pyoderma gangrenosum (n=33), and acne (n=24). Among all the cases, 13 genotypes were confirmed, and 47 variations involved amino acid p.E250. Steroid and/or biological agents were used in most patients. Conclusions: PAPA syndrome should be suspected in children with recurrent pyogenic sterile arthritis, and an early diagnosis could be achieved by genetic test. Targeted treatment with biological agent may control the symptoms effectively. Biological agents can control symptoms of this disorder effectively.

LncRNA SNHG20 promoted the proliferation of glioma cells via sponging miR-4486 to regulate the MDM2-p53 pathway.

OBJECTIVE: Evidence demonstrated the critical role of long noncoding RNAs (lncRNAs) in the initiation and development of human cancers. LncRNA small nucleolar RNA host gene 20 (SNHG20) was recently reported to promote the progression of several cancers; however, the function of SNHG20 in glioma has not been characterized. PATIENTS AND METHODS: Differential expression of SNHG20 in glioma tissues and cell lines was analyzed by RT-qPCR. Cell Counting Kit-8 (CCK-8) assay was performed to detect the cell viability. The targets prediction was analyzed with the TargetScan database. Western blot was performed to check the protein expression. RESULTS: SNHG20 was overexpressed in glioma tissues and cell lines. Down-regulation of SNHG20 suppressed the proliferation, migration and induced apoptosis of glioma cells. Molecular studies uncovered that SNHG20 acted as a competing endogenous RNA (ceRNA) to sponge the expression of miR-4486. MiR-4486 was down-regulated in glioma tissues and significantly inversely correlated with the expression of SNHG20. Restoration of miR-4486 remarkably attenuated the promotion effect of SNHG20 on the growth of glioma cells. Further study revealed that miR-4486 targeted the E3 ubiquitin ligase mouse double minute 2 (MDM2) and negatively regulated the expression of MDM2. Down-regulation of MDM2 by miR-4486 increased the abundance of p53 in glioma cells. CONCLUSIONS: We identified the functional mechanism by which SNHG20 modulated the malignancy of glioma cells via targeting the miR-4486/MDM2/p53 pathway. Interrupting the expression of SNHG20 might be a novel strategy to suppress the progression of glioma.

Transplantation of a reverse facial artery pedicled submandibular gland in a dog.

This study was performed to evaluate the possibility of using reverse facial artery pedicled submandibular gland transfer for treating keratoconjunctivitis sicca. A reverse facial artery pedicled submandibular gland, including the duct, from a beagle dog was transplanted into the temporal region in the same animal. 99mTc pertechnetate scintigraphy was used to monitor graft viability, and the grafted glands were examined histologically 12 weeks after transplantation. Postoperative 99mTc pertechnetate scintigraphy demonstrated viable salivary gland tissue in the transplanted region. Histologically, some of the acinar cells in the graft had atrophied. The transplantation of a reverse facial artery pedicled submandibular gland was successful and may be a simple, reliable treatment for patients with keratoconjunctivitis sicca.

Effect of downregulated lncRNA NBAT1 on the biological behavior of glioblastoma cells.

OBJECTIVE: To investigate the expression of lncRNA neuroblastoma associated transcript 1 (NBAT1) in human glioma cell lines and its underlying mechanism. Effect of NBAT1 on biological behaviors of T98 and U87 cells are also explored. PATIENTS AND METHODS: The mRNA expressions of NBAT1 in 48 cases of glioblastoma tissues and 30 cases of normal brain tissues were accessed by Real-time fluorescence quantitative PCR (RT-PCR). The relationship between mRNA expression of NBAT1 and tumor size, malignancy, and prognosis were analyzed. Effects of NBAT1 on the proliferation of glioblastoma T98 and U87 cells were determined by CCK-8 assay and colony formation assay, respectively. RESULTS: NBAT1 expressions in glioblastoma tissues were lower than those in normal brain tissues, which was negatively correlated with malignancy degree (p<0.01). Protein levels of Akt were decreased in T98 and U87 cells transfected with si-NBAT1. Meanwhile, proliferation abilities of T98 and U87 cells transfected with si-NBAT1 were significantly decreased as well (p<0.01), which were reversed by transfection of si-Akt. CONCLUSIONS: Upregulated NBAT1 inhibits proliferation of T98 and U87 cells via regulating Akt, indicating that NBAT1 may be related to the malignancy and prognosis of gliomas.

Expression and pathological relevance of inducible nitric oxide synthase in osteosarcoma of the jaws.

The aim of the present study was to examine the expression of inducible nitric oxide synthase (iNOS) in osteosarcoma of the jaw, and its relationship with tumour angiogenesis and clinicopathological characteristics. Streptavidin peroxidase immunohistochemical staining was used to detect the expression level of iNOS and CD34 in paraffin-embedded samples from 25 patients. Osteosarcoma of the jaw was associated with overexpression of iNOS, which correlated with tumour microvessel density (MVD). iNOS expression correlated with the size, pathological grade and clinical stage of the osteosarcoma, and also with clinicopathological characteristics such as primary occurrence or recurrence of tumours. There was no correlation with metastasis. iNOS may promote tumour angiogenesis in osteosarcoma of the jaw, and so may represent an important target in anti-tumour therapy.

Potential confounding factors in measurement of specific cell-free seminal mRNAs and microRNAs derived from human reproductive organs.

Cell-free seminal RNA (cfs-RNA) is mixed transcripts derived from male reproductive organs, and is potential biomarker for the research and diagnosis of male reproductive-related diseases. However, some clinical factors, including age, asymptomatic Ureaplasma urealyticum (UU) infection, scrotal heat stress, abstinence period, and the storage condition of semen samples, may interfere with sperm parameters and the measurement of seminal biomarkers. Accordingly, this study was designed to evaluate the effect of above clinical factors on the measurement of cfs-RNA, aiming to lay a foundation for its research use and potential clinical application. Semen samples were collected according to the selected clinical factors. Cell-free seminal plasma was obtained by centrifugation and total RNA was extracted with TRIzol LS. Selective male reproductive organ-specific cfs-mRNAs and cfs-miRNAs were quantified by quantitative real-time PCR. The concentration and total amount of cfs-mRNAs and cfs-miRNAs in one ejaculate were calculated and compared. ACTB, DDX4 (testis-specific), WFDC9 (epididymis-specific), and miR-514a-3p (testis-specific) significantly increased after scrotal heat stress. SEMG1 (seminal vesicle-specific) showed declining tendency with the prolonged abstinence period. Age, asymptomatic UU infection, and the storage condition showed no significant impact on the measurement of cfs-RNA. These results indicate that scrotal heat stress significantly interfere with the selected cfs-RNA derived from the testis and epididymis, and abstinence period may affect the yield of cfs-mRNA from seminal vesicle, while other clinical factors has no significant impact on the measurement. Thus, heat exposure and abstinence period should be considered for the cfs-RNA measurement in its research or clinical application.

The rs5934505 single nucleotide polymorphism (SNP) is associated with low testosterone and late-onset hypogonadism, but the rs10822184 SNP is associated with overweight and obesity in a Chinese Han population: a case-control study.

Low testosterone is associated with late-onset hypogonadism (LOH) and obesity. Recently, studies have shown that four single nucleotide polymorphisms (SNPs), rs12150660, rs727428, rs5934505, and rs10822184, are associated with testosterone levels in populations of European descent. Therefore, we investigated whether the SNP loci are related to low testosterone, LOH, or obesity in a Chinese Han population. Ruling out co-morbidities, DNA was prepared from 409 men (aged 40-65 years) with low serum testosterone (defined as total testosterone <11.6 nmol/L) and 1 : 1 normal controls (matched age, body mass index (BMI), and the same living area) who were selected from 6898 males. According to the same standards, 310 men with LOH and 1 : 1 normal controls were selected from 6898 males. Excluding the cases with an unreliable sequencing result, genetic analyses were performed. The minor allele frequencies of the SNP loci rs12150660, rs727428, rs5934505, and rs10822184 were 0.1%, 44.6%, 18.7%, and 38.9%, respectively. rs5934505 was associated with the serum total testosterone and calculated free testosterone (CFT) levels (p = 0.045 and p = 0.021). rs5934505 (C>T) was associated with an increased risk of low total testosterone, low CFT, and LOH and adjusted for other factors, with an odds ratio (OR) of 2.01 (1.34-3.01), 2.14 (1.42-3.20), and 1.64 (1.04-2.58). rs10822184 was significantly correlated with weight and BMI (p = 0.035 and p = 0.027). rs10822184 (T>C) was associated with an increased risk of overweight and obesity. We adjusted for other factors, with odds ratios (ORs) of 1.94 (1.36-2.78) and 1.56 (1.00-2.43). In summary, our study provided convincing evidence that rs5934505 (C>T) was associated with the risk of low testosterone and LOH in Chinese populations. We were the first to find that rs10822184 (T>C) was significantly correlated with the risk of overweight and obesity in Chinese populations. However, further large and functional studies are warranted to confirm our findings.

Defibrillation shocks produce different effects on Purkinje fibers and ventricular muscle: implications for successful defibrillation, refibrillation and postshock arrhythmia.

OBJECTIVES: To understand the mechanisms of postdefibrillation arrhythmias and failed defibrillation, we studied the cellular effects of high voltage shocks on different cardiomyocytes in the dog. BACKGROUND: The causes of postdefibrillation arrhythmias and unsuccessful defibrillation are not clear. METHODS: High voltage shocks with voltage differentials of 9.3 to 97.6 V/cm were delivered to isolated canine papillary muscles with attached Purkinje fibers. Transmembrane potentials were recorded simultaneously from the Purkinje fiber and the ventricular muscle using standard microelectrode techniques. RESULTS: After delivery of high voltage shocks, significant depolarization and rapid firing were observed in Purkinje fibers. The maximal rate of the rapid firing in the Purkinje fibers correlated with shock intensity (r = 0.69, p < 0.05). In contrast, in ventricular muscle, only slight depolarization and a transient refractory state were observed after the shock. The incidence of the refractory state was correlated with both the shock intensity and the rate of the rapid firing in the Purkinje fiber (r = 0.89 and 0.74, p < 0.01 and 0.05, respectively). Propranolol at a concentration sufficient for complete beta-blockade (1 mg/liter) did not change the tissue response to shocks but suppressed or abolished the shock-induced rapid firing of Purkinje fibers at a higher concentration (3 mg/liter). Blockade of the slow calcium channel with verapamil (400 micrograms/dl) did not alter the responsiveness of the preparation to shocks. CONCLUSION: These results indicate that high voltage shocks induce different responses in Purkinje fibers and ventricular muscle. The shock-induced rapid firing in the Purkinje fiber may contribute to postshock arrhythmias and possibly refibrillation in some cases. The shock-induced transient refractory state in the ventricular muscle may prevent the ventricle from responding to the rapid firing and thus may decrease the incidence of postshock arrhythmias.

Electrophysiologic substrate associated with pacing-induced heart failure in dogs: potential value of programmed stimulation in predicting sudden death.

To investigate the possible mechanisms of sudden death and the potential role of electrophysiologic testing in congestive heart failure, this study evaluated the electrophysiologic substrate in a model of heart failure induced by rapid pacing. Seventeen mongrel dogs underwent cardiac pacing at 220 to 240 beats/min for 5 weeks (paced group) and 11 other dogs served as a sham-operated control group. Rapid pacing of the right ventricle produced clinical and hemodynamic features of congestive heart failure. Dogs in the paced group had prolonged cardiac conduction time as reflected by longer epicardial activation time (36.1 +/- 2.4 vs. 30.8 +/- 0.8 ms, p less than 0.05). The ventricular effective refractory period was significantly prolonged after the development of heart failure (141 +/- 4 vs. 177 +/- 5 ms, p less than 0.01, at a basic pacing cycle length of 300 ms), whereas no significant change was found in the control group (140 +/- 4 vs. 145 +/- 4 ms, p = NS). The prolongation of the ventricular effective refractory period correlated with an increase in left ventricular end-diastolic pressure (r = 0.55, p less than 0.001) and the ventricular effective refractory period correlated inversely with cardiac index (r = -0.49, p less than 0.025). The rest membrane potential of ventricular muscle was less negative in the paced group compared with the control group (-80.7 +/- 2.2 vs. -85.6 +/- 2.2 mV, p less than 0.05). Intracellularly recorded action potential duration of ventricular muscle was longer in the paced than in the control group (236 +/- 9.8 vs. 198.9 +/- 2.6 ms, p less than 0.01), action potential duration at 90% repolarization).(ABSTRACT TRUNCATED AT 250 WORDS)