A Case Report of Brucellosis and the Differential Diagnosis of Lymphoma and Tuberculosis

ABSTRACT In this paper, we present a male patient from Xinjiang with fever of unknown origin and significant weight loss for more than 1 month. He was admitted to hospital with negative Rose Bengal test (RBT) and decreased leucocyte count. Ultrasound revealed splenomegaly and abdominal computed tomography, which showed multiple hypodense splenic nodules. The patient was suspected of lymphoma or tuberculosis. Pathological biopsy suggested brucellosis infection following splenectomy. Anti-Brucella treatment was effective and his temperature gradually returned to normal. During the follow-up, the patient's RBT result turned to positive and he was instructed to continue the anti-Brucella drug regimen. His temperature, weight, white blood cell count, other laboratory examinations, and imaging findings all returned to normal during the 6-month follow-up after the treatment.

The effects of threonine on performance parameters, carcass traits, visceral organ indices and serum biochemical parameters of Linwu ducks, aged 4 to 8 weeks

This study was conducted to establish dietary threonine (Thr) levels for Linwu ducks, aged 4 to 8 wk. Experimental diets formulated to contain 0.56, 0.61, 0.66, 0.71, 0.76, and 0.81% Thr fed to Linwu ducks. A total of 360 healthy Linwu female ducks with similar body weight (1183.89±3.83 g) were randomly divided into six groups, with five replicates in each group, and 12 ducks in each replicate. Samples were collected at 8 wk for the determination of growth performance, carcass traits, visceral organ indices, and serum biochemical parameters. As a result of this study, Thr level had no significant influence on the final weight, the daily gain, feed/gain ratio, and average daily intake (p>0.05). Similarly, there were no significant effects of dietary Thr on carcass traits and visceral organ indices (p>0.05). The pancreatic index was highest among all the treatments when the dietary Thr level was 0.66%. The different dietary Thr levels had no significant effect (p>0.05) on the concentration of total protein (TP), triglyceride (TG), total cholesterol (TC), glucose (GLU), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and glutathione (GSH). However, the serum MDA concentration in the 0.66% treatment was lower (P=0.068) than in the other treatments. In conclusion, Thr at 0.66% concentration may have an antioxidant activity and exert positive effect on Linwu ducks.

The effects of threonine on performance parameters, carcass traits, visceral organ indices and serum biochemical parameters of Linwu ducks, aged 4 to 8 weeks

This study was conducted to establish dietary threonine (Thr) levels for Linwu ducks, aged 4 to 8 wk. Experimental diets formulated to contain 0.56, 0.61, 0.66, 0.71, 0.76, and 0.81% Thr fed to Linwu ducks. A total of 360 healthy Linwu female ducks with similar body weight (1183.89±3.83 g) were randomly divided into six groups, with five replicates in each group, and 12 ducks in each replicate. Samples were collected at 8 wk for the determination of growth performance, carcass traits, visceral organ indices, and serum biochemical parameters. As a result of this study, Thr level had no significant influence on the final weight, the daily gain, feed/gain ratio, and average daily intake (p>0.05). Similarly, there were no significant effects of dietary Thr on carcass traits and visceral organ indices (p>0.05). The pancreatic index was highest among all the treatments when the dietary Thr level was 0.66%. The different dietary Thr levels had no significant effect (p>0.05) on the concentration of total protein (TP), triglyceride (TG), total cholesterol (TC), glucose (GLU), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and glutathione (GSH). However, the serum MDA concentration in the 0.66% treatment was lower (P=0.068) than in the other treatments. In conclusion, Thr at 0.66% concentration may have an antioxidant activity and exert positive effect on Linwu ducks.(AU)

Serum total bilirubin levels are negatively correlated with metabolic syndrome in aged Chinese women: a community-based study.

We evaluated serum total bilirubin levels as a predictor for metabolic syndrome (MetS) and investigated the relationship between serum total bilirubin levels and MetS prevalence. This cross-sectional study included 1728 participants over 65 years of age from Eastern China. Anthropometric data, lifestyle information, and previous medical history were collected. We then measured serum levels of fasting blood-glucose, total cholesterol, triglycerides, and total bilirubin, as well as alanine aminotransferase activity. The prevalence of MetS and each of its individual component were calculated per quartile of total bilirubin level. Logistic regression was used to assess the correlation between serum total bilirubin levels and MetS. Total bilirubin level in the women who did not have MetS was significantly higher than in those who had MetS (P<0.001). Serum total bilirubin quartiles were linearly and negatively correlated with MetS prevalence and hypertriglyceridemia (HTG) in females (P<0.005). Logistic regression showed that serum total bilirubin was an independent predictor of MetS for females (OR: 0.910, 95%CI: 0.863-0.960; P=0.001). The present study suggests that physiological levels of serum total bilirubin might be an independent risk factor for aged Chinese women, and the prevalence of MetS and HTG are negatively correlated to serum total bilirubin levels.

Molecular characterization of the gene checkpoint homolog 1 in Daphnia carinata during different reproductive phases.

Full-length cDNA of the gene checkpoint homolog 1 (Chk1) was cloned from Daphnia carinata and designated DcarChk1. DcarChk1 cDNA was 1817 bp in length and encoded a 497-amino acid polypeptide. Phylogenetic analyses revealed that DcarChk1 was most closely related to Chk1 of Daphnia pulex, followed by homologous genes of insects. Expression of DcarChk1 was higher in adult Daphnia than in larvae, and significantly higher in males than females, as determined by real-time polymerase chain reaction analysis. Using whole-mount in situ hybridization techniques, DcarChk1 in parthenogenetic females was found to be expressed mainly on the head surface, capillus, and carapace valve edge. In contrast, in sexual females, DcarChk1 was expressed mainly in the joint of the second antenna, and in the thoracic limbs and capillus. These results suggest that DcarChk1 plays a significant role in both the growth and development, as well as in regulating reproductive plasticity, in D. carinata.

Overexpression of Cathepsin L is associated with gefitinib resistance in non-small cell lung cancer.

Lung cancer, the most common malignancy, is still the leading cause of cancer-related death worldwide. Non-small-cell lung cancer (NSCLC) accounts for 80 % of all lung cancers. Recent studies showed Cathepsin L (CTSL) is overexpressed in various cancerous tissues; however, the association between CTSL expression and EGFR-TKI resistance remains unknown. In this study, we investigated the expression of CTSL in lung cancer specimens and matched normal tissues by quantitative real-time PCR and IHC. The functional role of CTSL in resistant PC-9/GR cell line was investigated by proliferation and apoptosis analysis compared with control PC-9 cells. Our results found that the level of CTSL expression was higher in NSCLC tissues compared with matched normal adjacent tissue samples, and CTSL was more highly expressed in PC-9/GR cells compared to PC-9 cells. Knocking-down of CTSL in PC-9/GR cells could decrease cell proliferation and potentiate apoptosis induced by gefitinib, suggesting CTSL may contribute to gefitinib resistance in NSCLC. CTSL might be explored as a candidate of therapeutic target for modulating EGFR-TKI sensitivity in NSCLC.

Improvements in neuroelectrophysiological and rear limb functions in rats with spinal cord injury after Schwann cell transplantation in combination with a C5a receptor antagonist.

We measured the effect of Schwann cell transplantation and complement factor 5a (C5a) receptor antagonist on nerve function recovery in rats with spinal cord injury. Experimental spinal cord injury was induced in eighty Wistar rats and these were randomly divided into four treatment groups: culture medium and saline injection (control group), Schwann cell injection (cell transplantation group), C5a receptor antagonist injection (C5a receptor antagonist group), and both Schwann cell and C5a receptor antagonist injections (combination group). Rear limb functional recovery was assessed 1, 2, 4, 6, and 8 weeks after the spinal cord injury with the tilt table test and the Basso, Beattie, Bresnahan scale. Sex-determining region Y (SRY) gene expression was measured at week 4 and horseradish peroxidase (HRP) labeling was used at week 8 to further assess the recovery of neuroelectrophysiological functions. The rear limb functional assessment showed that the combination group had better outcomes than the cell transplantation and C5a receptor antagonist groups. All treatment groups had better outcomes than control. Only the cell transplantation and combination groups showed SRY expression. The number of HRP-positive nerve fibers in the different groups ranked as follows: combination group > cell transplantation and C5a receptor antagonist > control. The refractory period and amplitude of the induced potential in the combination group were significantly greater than in the other three groups. These results suggest that the combination of Schwann cell transplantation and the C5a receptor antagonist enhances the regeneration of injured synapses and improves limb function and electrophysiology.

Characterization of ESTs from black locust for gene discovery and marker development.

Black locust (Robinia pseudoacacia L.) is an ecologically and economically important species. However, it has relatively underdeveloped genomic resources, and this limits gene discovery and marker-assisted selective breeding. In the present study, we obtained large-scale transcriptome data using a next-generation sequencing platform to compensate for the lack of black locust genomic information. Increasing the amount of transcriptome data for black locust will provide a valuable resource for multi-gene phylogenetic analyses and will facilitate research on the mechanisms whereby conserved genes and functions are maintained in the face of species divergence. We sequenced the black locust transcriptome from a cDNA library of multiple tissues and individuals on an Illumina platform, and this produced 108,229,352 clean sequence reads. The high-quality overlapping expressed sequence tags (ESTs) were assembled into 36,533 unigenes, and 4781 simple sequence repeats were characterized. A large collection of high-quality ESTs was obtained, de novo assembled, and characterized. Our results markedly expand the previous transcript catalogues of black locust and can gradually be applied to black locust breeding programs. Furthermore, our data will facilitate future research on the comparative genomics of black locust and related species.

New compound heterozygous mutations of p. Thr101Ilefs 2 and p. Thr306Ale in a child from a Chinese family with 17α-hydroxylase/17, 20-lyase deficiency.

We determined whether a child with 17α-hydroxylase/17, 20-lyase deficiency possessed the sex-determining region (SRY) gene, and examined the mutations present in the CYP17A1 gene that led to 17α-hydroxylase/17, 20-lyase deficiency. In the child, karyotype analysis was performed and polymerase chain reaction analysis and electrophoretic techniques were used to identify the SRY gene. A total of 50 normal individuals were included as a control group. Polymerase chain reaction and DNA sequencing were used to identify CYP17A1 gene mutations in all samples. The karyotype of the child was 46, XY, which was inconsistent with her social sex, SRY was positive, and a compound heterozygous mutation p. Thr101Ilefs*2 in exon 2 and p. Thr306Ale in exon 5 were identified in the CYP17A1 gene. These mutations were inherited from her parents. In the 20 normal individuals, these mutations were not identified. In the child, sex reversal may have been caused by CYP17A1 mutations. The compound heterozygous mutation of p. Thr101Ilefs*2 and p. Thr306Ale is a new gene mutation of 17α-hydroxylase/17, 20-lyase deficiency.

Identification and evaluation of endogenous control genes for use in quantitative RT-PCR during wheat (Triticum aestivum L.) grain filling.

The use of appropriate reference genes is essential for the generation of accurate and biologically meaningful results from quantitative real-time PCR (qRT-PCR) analysis. However, studies have found that the expression of most commonly used reference genes is not always independent of the tissues, treatments, or developmental stages studied. geNormPlus, NormFinder, and BestKeeper, were applied and the expression stability of nine candidate genes was evaluated in different data sets during wheat grain development. Varying degrees of diversity in either single or multiple reference genes were observed among the results generated from the different computer programs, parameters, and data sets. Therefore, the reliability of identified reference genes in the flag leaf and the complete set of samples was estimated by monitoring the expression dynamics of three NAM genes (TaNAM-A1, TaNAM-B1, and TaNAM-B2). The results suggest that a single control gene identified by geNormPlus for use with the complete set of samples, and multiple reference genes selected by geNormPlus and NormFinder exclusively for the flag leaf outperformed others owing to the consistent results with previous analyses of these genes, which were normalized against a verified single control gene. Given the limit of NormFinder in gene numbers of multiple reference genes, robust quantification can be achieved by normalizing against Ta27922 or multiple reference genes chosen by geNormPlus for individual tissues.

Anti-prostate cancer effects of CTL cell induction in vitro by recombinant adenovirus mediated PSMA/4-1BBL dendritic cells: an immunotherapy study.

This study aimed to examine anti-prostate cancer immune response induced by dendritic cells (DCs) transduced with PSMA/4-1BBL recombinant adenoviruses in vitro. Ad-PSMA, Ad-4-1BBL, and Ad-GFP were transfected into DCs derived from peripheral blood of healthy volunteers. Ad-PSMA/4-1BBL-DC, Ad-PSMA-DC, Ad-4-1BBL-DC, Ad-GFP-DC, and normal-DC, PSMA and 4-1BBL protein levels in DCs were detected by western blot. IL-12, IFN-γ and IL-10 were measured by ELISA. Mixed lymphocyte reaction and the cytotoxicity of each group targeted to LNCap, Du145, and 22RV prostate cancer cells were determined by CCK-8 assay. PSMA and 4-1BBL protein could express on DC successfully, the IL-12 supernatant content (134.29 ± 2.22 pg) was higher than others (P < 0.05). The ability to stimulate autologous T lymphocyte proliferation in the co-transfection group was higher than others (P < 0.05). When the DCs were co-cultured with CTLs, the PSMA/4-1BBL-DC-CTL group showed the highest content of IFN-γ (1176.10 ± 14.37pg/5 x 10(6) cells), but the lowest IL-10 content (75.14 ± 2.01 pg/5 x 10(6) cells) (P < 0.05), and the strongest anti-tumor effect when the effector to target ratio was 40:1, along with a higher killing ratio of LNCap cells than others (P < 0.05). Overall, Mature DCs transfected with Ad-PSMA/4- 1BBL not only showed high secretion of IL-12, but also induced CTLs to stimulate and enhance the killing effect of PSMA specific effector cells to PSMA positively expressing prostate cancer cells. Furthermore, the DCs infected with two kinds of tumor-associated antigens would induce more effective tumor-specific CTL induction.

Effect of ultraviolet A exposure on transport of compatible organic osmolytes in human lens epithelial cells.

Compatible organic osmolytes, such as betaine, myoinositol, and taurine, are involved in antioxidant defense, protein stabilization, and stress responses. This osmolyte strategy requires the expression of specific osmolyte transporters such as betaine (BGT-1), myoinositol (SMIT), and taurine (TAUT). In contrast to the kidney, keratinocytes, and neural cells, few studies have examined osmolytes in human lens epithelial cells (HLECs). We examined the expression of mRNA specific for BGT-1, SMIT, and TAUT in HLECs. In comparison to normoosmotic (305 mOsM) controls, there was a 3-5-fold time-dependent reaction of BGT-1, SMIT, and TAUT mRNA levels in HLECs exposed to hyperosmotic stress (405 mOsM). Maximal responses were obtained for BGT-1, SMIT, and TAUT mRNA expression after 3, 24 and 9 h of hyperosmotic exposure, respectively. This expression was correlated with increased osmolyte uptake. In contrast, hypoosmotic (205 mOsM) stimulation led to a significant efflux of osmolytes. Exposure to ultraviolet A (340-400 nm) radiation significantly stimulated osmolyte uptake. Increased osmolyte uptake was associated with upregulation of mRNA steady-state levels for osmolyte transporters in irradiated cells. These results demonstrate that ultraviolet A radiation leads to the accumulation of compatible organic osmolytes in HLECs as hyperosmotic pressure, which can maintain cellular environmental homeostasis.

Analysis of prognostic factors for infantile rotavirus infection.

In this study, we investigated the prognostic factors of infantile rotavirus (RV) infection. A total of 102 infants with RV enteritis were divided into 2 groups according to the standards of improvement and cure at the time of discharge from the hospital: improvement group (N = 58; 47 males and 11 females with an average age of 15.19 ± 5.03 months) and the cure group (N = 44; 34 males and 10 females with an average age of 10.02 ± 4.92 months). Both groups were phlebotomized for the detection of serum glutamic oxaloacetic aminotransferase, creatine kinase-MB, and lactate dehydrogenase. Prognostic factors and clinical data were analyzed by univariate and multivariate logistic analysis. Among the 102 cases of RV infection, 58 were cured and 44 were improved. Univariate analysis showed that the 2 groups were significantly different in age, feeding pattern, concentrations of serum glutamic oxaloacetic aminotransferase, creatine kinase-MB, and lactate dehydrogenase, and central nervous system damage. Logistic regression analysis showed that age, feeding, and central nervous system damage were significant independent prognostic factors for RV enteritis (P < 0.05). There were no statistical differences in gender, course of disease, and respiratory infection (P < 0.05). Both myocardial and hepatic damages presented a temporary feature in the infants and had no significant influence on prognosis. Age, feeding pattern, and central nervous system damage are significant independent prognostic factors for RV infection. These factors should be carefully considered in clinical practice.

Retrospective analysis of pathologic nipple discharge.

The cause of pathologic nipple discharge is mainly benign lesions, but there is still a possibility of malignancy. Pathologic nipple discharge may be the only or the first symptom of breast cancer. This study aimed to investigate the clinical factors associated with lesions in patients with pathologic nipple discharge using a retrospective analysis of clinical data in 207 cases. The univariate analysis showed that age >50 years, breast lumps, or breast calcifications were risk factors associated with breast cancer in nipple discharge patients (P < 0.05). Discharge characteristics, duration of disease, and identification of lesions had no clear clinical significance (P > 0.05). The multivariate analysis also showed that age >50 years, breast lumps, and breast calcifications were risk factors associated with breast cancer in nipple discharge patients (P < 0.05). Age, breast lumps, and breast calcifications had important clinical significance in identification of benign and malignant nipple discharge.

Association of vitamin D receptor BsmI gene polymorphism with risk of low bone mineral density in post-menopausal women: a meta-analysis.

The vitamin D receptor BsmI gene polymorphism is reportedly associated with low bone mineral density (BMD) in postmenopausal women, but results from previous studies are conflicting. In the present study, we investigated the association between this polymorphism and the risk of low BMD through a meta-analysis of published studies. A literature search of the Pubmed, Embase, and CNKI databases from inception through July 2013 was conducted. The meta-analysis was performed using the STATA 12.0 software. Crude odds ratios with 95% confidence intervals were used to assess the strength of any association. Eleven case-control studies were included for a total of 1468 low BMD cases and 2177 healthy controls. No significant variation in low BMD risk was detected in any of the genetic models. Further stratified analyses were performed to examine the effect of ethnicity. In the subgroup analysis, no significant association was found in Caucasians and in Asians. The meta-analysis results suggest that the BsmI polymorphism is not associated with low BMD risk in postmenopausal women.

Analysis of intervertebral disc-related genes.

Intervertebral disk disease is a common clinical disorder manifested by pain, ataxia, paresis, motor paralysis, and sensorimotor paralysis. The clinical features, diagnosis, and treatment of cervical and thoracolumbar disk disease have been unclear until now. In this study, some differentially expressed genes were identified, and a network was constructed based on these genes. Through the statistical analysis of nodes and the contrast of 2 more connectivity nodes, it was found that the nodes in the network are in an important position and play key roles. Several of these genes, including MAP2K6, MAP2K3, and MAPK14, belong to the MAP kinase family, and several genes, including RHOBTB2, RHOQ, and RHOH, belong to the RHO family. Therefore, we hypothesize that the development of intervertebral disk disease is related to MAP and RHO family proteins.

Correlation analysis between starch properties and single nucleotide polymorphisms of waxy genes in common rye (Secale cereale L.).

To understand the relationships between single nucleotide polymorphisms (SNPs) in the waxy gene and starch parameters in common rye, we performed sequence characterization, enzyme activity testing, amylopectin/amylose ratio evaluation, starch property testing, and correlation analysis. Specific primers were used to clone waxy from 20 rye cultivars. Sequence analysis showed that waxy was 2852 bp, including 11 exons, and sequence similarity across the 20 cultivars was over 98%. The Waxy protein showed >95% similarity with those from wheat, rice, and barley, the closest genetic relationship being with wheat Wx-A type. Waxy had multiple SNPs, most of which were located in the exons. Amino acid variants were found to be mainly distributed in the catalytic domain in an imbalanced state. Multi-factor correlation analysis revealed significant correlation among starch pasting parameters in rye flour. The Waxy protein activity was significantly negatively correlated with the amylose content and amylopectin/amylose ratio. However, pasting parameters, Waxy enzyme activity, and amylopectin/amylose content ratio were not correlated. The correlation of SNPs, the key catalytic site of Waxy, with starch parameters and enzyme activity suggested that both starch pasting parameters and Waxy protein activity were influenced by No. 260 amino acid (aa). Further, the 141 and 152 aa loci were found in the enzyme-catalyzing domain of Waxy. Interestingly, Waxy enzyme activity was also influenced by the 363 aa locus in the pliable region. These results provide important theoretical regarding the high-throughput quality identification of noodle starch, functional studies, directional selection, and molecular markers of wheat Wx subunits.

Early responses of the STAT3 pathway to platinum drugs are associated with cisplatin resistance in epithelial ovarian cancer.

Cisplatin resistance remains one of the major obstacles when treating epithelial ovarian cancer. Because oxaliplatin and nedaplatin are effective against cisplatin-resistant ovarian cancer in clinical trials and signal transducer and activator of transcription 3 (STAT3) is associated with cisplatin resistance, we investigated whether overcoming cisplatin resistance by oxaliplatin and nedaplatin was associated with the STAT3 pathway in ovarian cancer. Alamar blue, clonogenic, and wound healing assays, and Western blot analysis were used to compare the effects of platinum drugs in SKOV-3 cells. At an equitoxic dose, oxaliplatin and nedaplatin exhibited similar inhibitory effects on colony-forming ability and greater inhibition on cell motility than cisplatin in ovarian cancer. Early in the time course of drug administration, cisplatin increased the expression of pSTAT3 (Tyr705), STAT3α, VEGF, survivin, and Bcl-XL, while oxaliplatin and nedaplatin exhibited the opposite effects, and upregulated pSTAT3 (Ser727) and STAT3ß. The STAT3 pathway responded early to platinum drugs associated with cisplatin resistance in epithelial ovarian cancer and provided a rationale for new therapeutic strategies to reverse cisplatin resistance.

Early responses of the STAT3 pathway to platinum drugs are associated with cisplatin resistance in epithelial ovarian cancer

Cisplatin resistance remains one of the major obstacles when treating epithelial ovarian cancer. Because oxaliplatin and nedaplatin are effective against cisplatin-resistant ovarian cancer in clinical trials and signal transducer and activator of transcription 3 (STAT3) is associated with cisplatin resistance, we investigated whether overcoming cisplatin resistance by oxaliplatin and nedaplatin was associated with the STAT3 pathway in ovarian cancer. Alamar blue, clonogenic, and wound healing assays, and Western blot analysis were used to compare the effects of platinum drugs in SKOV-3 cells. At an equitoxic dose, oxaliplatin and nedaplatin exhibited similar inhibitory effects on colony-forming ability and greater inhibition on cell motility than cisplatin in ovarian cancer. Early in the time course of drug administration, cisplatin increased the expression of pSTAT3 (Tyr705), STAT3α, VEGF, survivin, and Bcl-XL, while oxaliplatin and nedaplatin exhibited the opposite effects, and upregulated pSTAT3 (Ser727) and STAT3β. The STAT3 pathway responded early to platinum drugs associated with cisplatin resistance in epithelial ovarian cancer and provided a rationale for new therapeutic strategies to reverse cisplatin resistance.

Genetic variants on 17q21 are associated with asthma in a Han Chinese population.

A genome-wide study has shown an association between SNPs located on 17q21 and asthma. Such associations have been identified in several populations, but little is known about the Han Chinese population. We conducted a case-control study in a Han Chinese population to investigate the relationship between SNPs located on 17q21 and asthma; 241 asthmatic patients and 212 healthy controls were recruited from the outpatient clinics of the Nanfang Hospital, Guangdong Province, southern China. We genotyped six SNPs (rs8067378, rs8069176, rs2305480, rs4795400, rs12603332, and rs11650680) located on 17q21 with the Sequenom MassARRAY iPLEX platform. For two of these six loci (rs2305480 and rs8067378), there was evidence of association with asthma, and there was a weak association of asthma with rs8069176. We confirm that genetic variants on 17q21 are associated with asthma in the Han Chinese population.