[Clinical observation of different reperfusion methods in patients with acute ischemic stroke with atrial fibrillation within 4.5 hours from onset].

Objective: To detect the ideal treatment for acute ischemic stroke (AIS) patients with atrial fibrillation (AF) within 4.5 hours from onset. Methods: A total of 95 AIS patients with AF was retrospectively analyzed from April 2014 to January 2019. Thirty patients (group A) were treated with endovascular treatment directly, 35 (group B) patients were treated with intravenous recombinant tissue plasminogen activator (rtPA) followed by endovascular treatment, and 30 (group C) patients were treated with intravenous rtPA only. There were no significant differences among the groups in baseline data as age, gender, underlying diseases, medication, National Institutes of Health Stroke Scale (NIHSS) score, time from onset to treatment. Modified thrombolysis in cerebral infarction (mTICI), Symptomatic hemorrhagic transformation (SICH), 90 d prognosis of modified Rankin Scale (mRS) and death were compared. Results: Recanalization (mTICI≥2b) was similar in group A and B (70.0% vs. 68.6%, P>0.05). SICH of group A (6.7%) was significantly lower than that of group B (31.4%, P<0.05), but similar with that of group C (13.7%, P>0.05). Prognosis (mRS≤2) was significantly better in group A (70.0%) than that in group B (37.1%) and group C (30.0%), both P<0.01. The mortality rate in group A (6.7%) was lower than that in group B (14.3%) and group C (20.0%) without statistically significant, both P>0.05. Conclusion: AIS patients with AF within 4.5 hours from onset should receive endovascular treatment directly.

[Analysis of abnormal results of thyroid B-ultrasonography in medical radiological workers in Nanjing, China].

Objective: To investigate the abnormal condition of thyroid B-ultrasonography in medical radiological workers and provides basis for health management of them. Methods: From January 1, 2018 to December 31, 1648 staff members exposed to medical radiation who participated in radiation health examination in all medical institutions in Nanjing to analyze their thyroid B-ultrasonography and compare the sex, age, length of service, blood pressure, blood glucose, smoking, drinking, professional and The post was analyzed statistically, and the influencing factors of thyroid B super anomaly were analyzed. Results: Among the 1 648 staff members, the abnormal rate of thyroid B-ultrasonography accounted for 39.4% (650/1648) , and the comparison of thyroid nodule, thyroid echo irregularity and thyroid cyst detection rate was statistically significant (P<0.01) . Compared with the normal group, the abnormal rate of thyroid B was statistically significant in different age groups, different injury working age groups, different posts, blood pressure and blood sugar (P<0.01) . The abnormal rate of thyroid B-ultrasound increased with the increase of age and working age (χ(2trend)=93.093、99.346, P<0.01) , especially the age over 40 and working age over 20 years increased the risk of abnormal thyroid B-ultrasound. Abnormal blood pressure and blood sugar were the risk of abnormal thyroid B-ultrasound (P<0.05) . Drinking alcohol could reduce the risk of abnormal thyroid B-ultrasound (P<0.05) . Conclusion: Medical radiation workers in Nanjing thyroid abnormalities are affected by age, working age, post, blood pressure, blood glucose, should strengthen protection, do a good job of comprehensive health management.

[Effect of Dishevelled 2 on apoptosis in rheumatoid arthritis fibroblast-like synoviocytes through inhibiting JAK-STAT pathway].

Objective: To investigate if Dishevelled 2 (DVL2) regulates the apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) via the JAK-STAT pathway. Methods: DVL2 overexpressed lentivirus was transfected into RA-FLS and the apoptosis rate was detected by flow cytometry. The effect of DVL2 on RA-FLS signaling pathway was detected by RNA-seq, and then the key genes were verified by RT-PCR. Results: Compared with the control group, DVL2 significantly increased the apoptosis rate of MH7A (3.2%±2.2% vs 25.7%±4.5%). RNA-seq results showed that DVL2 down-regulated the JAK-STAT pathway.The results of RT-PCR showed that DVL2 inhibited the gene expression of JAK2, Stat1, and Stat2; DVL2 still inhibited the gene expression of JAK2 and Stat2 but not Stat1 after TNF-α stimulation.DVL2 inhibited the gene expression of Bcl-xL, and the gene expression of Bcl-2 and Bcl-xL after TNF-α stimulation. Conclusion: DVL2 can increase the apoptosis rate of RA-FLS through inhibiting the JAK-STAT pathway and its downstream anti-apoptotic gene.

[Effects of nasogastric and percutaneous endoscopic gastrostomy tube feeding on the susceptibility of pulmonary infection in long-term coma patients with stroke or traumatic brain injury].

Objective: To investigate the effects of nasogastric tube (NGT) and percutaneous endoscopic gastrostomy (PEG) tube feeding on the susceptibility of pulmonary infection in long-term coma patients with stroke or traumatic brain injury. Methods: A total of 295 candidates who were in long-term coma after stroke or traumatic brain injury but without pulmonary infection and eligible for PEG catheterization were screened prospectively between January 2014 and February 2018. The patients were divided into PEG group (86 patients) and NGT group (209 patients) according to the choice of next-of-kin. Data related to the susceptibility of pulmonary infection were collected and analyzed in the two groups one month after the catheterization. Results: After follow-up for one month, compared with NGT group, patients in PEG group had a lower incidence of pulmonary infection (23.3% vs 37.8%, P=0.023), a later occurrence of pulmonary infection (average time: 21 days vs 13 days, P=0.034), and a less severe pulmonary infection (7.0% vs 12.9%, P=0.029). The following characteristics made patients more susceptible to pulmonary infection: age ≥ 70 years (HR=1.619, 95% CI 1.054-2.172), Charlson comorbidity index ≥ 2 points (HR=1.647, 95% CI 1.043-2.485), using of proton pump inhibitor ≥ 7 days (HR=1.725, 95% CI 1.214-2.738), and number of pressure ulcers ≥ 3 (HR=2.109, 95% CI 1.128-3.844). However, serum albumin concentration ≥35 g/L (HR=0.670, 95% CI 0.375-0.963) was a protective factor for pulmonary infections. The number of consistent pathogens cultivated from saliva, gastric juice and sputum simultaneously in NGT and PEG group was 35 strains (27.8%) and 8 strains (13.3%), respectively (P=0.029). The mortality of pulmonary infection was similar in the two groups (3.5% vs 4.3%, P=1.000), but the death due to pulmonary infection in the PEG group occur later (median time: 20 days vs 11 days, P=0.012). Conclusions: PEG feeding was a preferred nutrition way which could reduce the risk of pulmonary infection more effectively than NGT feeding which might favored a retrograde gastro-pulmonary route by which pathogens colonized in stomach migrated to respiratory tract. Patients with characteristics mentioned above had the susceptibility of pulmonary infection, thus risk assessment of pulmonary infection should be conducted before selecting the catheterization method.

[The health status of radiation workers and the analysis of crystalline lens results].

Objective: To understand the healthy status of radiation workers on the staff in Nanjing, and to analysis the relationship between abnormal crystalline lens and its influencing factors. Methods: We described physical indicators including blood pressure (BP) 、blood sugar (BG) 、thyroid B ultrasound、crystalline lens、chromosome and so on among 3 349 radiation workers on the staff in the year 2016 from Jan 1 to Dec 31, and the abnormal results of crystalline lens were analyzed statistically. Results: The rate of abnormal BP、BG、WBC、Thyroid B ultrasound、crystalline lens was 19.0%、2.2%、5.8%、30.0%、3.6% respectively; The rate of chromosome aberration was 0.1%, and the chromosomal micronuclei are all within normal range. With the rate of abnormal crystalline lens increasing in age and working years, statistical significance both existed in the trend; Compared to the lowest group, the risk of abnormal crystalline lens increased 3.86 times in ≥60 year old group and 3.16 times in ≥30 years working group; The risk of abnormal crystalline lens in nosocomial radation group was higher than non-medical group; There's no found in smoking and drinking alcohol increasing the risk. Morphologically, dot abnormal focused on 30~39 years old and 0~9 working years group, while lamellar abnormal concentrated upon ≥60 year old、≥30 working years group; Age and working-year were both the risk factors of lamellar abnormal; The risk of lamellar abnormal in nosocomial radiation group was significantly higher than non-medical group. Conclusion: Existing nisk foctions in the radiation work has a serious impact on several healthy physiology indicators, the more prominent was crystalline lens. Attention should be paid to eye protection and comprehensive health management.

Multiple variants in 5q31.1 are associated with systemic lupus erythematosus susceptibility and subphenotypes in the Han Chinese population.

BACKGROUND: A previous study provided evidence for a genetic association between PPP2CA on 5q31.1 and systemic lupus erythematosus (SLE) across multi-ancestral cohorts, but failed to find significant evidence for an association in the Han Chinese population. OBJECTIVES: To explore the association between this locus and SLE using data from our previously published genome-wide association study (GWAS). METHODS: Single-nucleotide polymorphisms (SNPs) rs7726414 and rs244689 (near TCF7 and PPP2CA in 5q31.1) were selected as candidate independent associations from a large-scale study in a Han Chinese population consisting of 1047 cases and 1205 controls. Subsequently, 3509 cases and 8246 controls were genotyped in two further replication studies. We then investigated the SNPs' associations with SLE subphenotypes and gene expression in peripheral blood mononuclear cells. RESULTS: Highly significant associations with SLE in the Han Chinese population were detected for SNPs rs7726414 and rs244689 by combining the genotype data from our previous GWAS and two independent replication cohorts. Further conditional analyses indicated that these two SNPs contribute to disease susceptibility independently. A significant association with SLE, age at diagnosis < 20 years, was found for rs7726414 (P = 0·001). The expression levels of TCF7 and PPP2CA messenger RNA in patients with SLE were significantly decreased compared with those in healthy controls. CONCLUSIONS: This study found evidence for multiple associations with SLE in 5q31.1 at genome-wide levels of significance for the first time in a Han Chinese population, in a combined genotype dataset. These findings suggest that variants in the 5q31.1 locus not only provide novel insights into the genetic architecture of SLE, but also contribute to the complex subphenotypes of SLE.

[Significance of monitoring the gradients between transcutaneous PCO(2) and end-tidal PCO(2) in patients with septic shock].

Objective: To investigate the significance of monitoring the gradients between transcutaneous PCO(2) and end-tidal PCO(2) [P(c-et)CO(2)] in patients with septic shock. Method: Thirty-five mechanically ventilated patients with early septic shock were enrolled as the study group and 18 non-septic shock patients with stable hemodynamics as the control group between May 2014 and October 2016. The patients with septic shock were treated by early goal-directed therapy (EGDT) within 6 hours since hospitalization. The differences of baseline level of P(c-et)CO(2) and arterial lactate concentration (LAC) between the two groups and the variations of these indexes after EGDT in the study group were compared respectively. Results: The baseline levels of P(c-et)CO(2) and LAC in patients with septic shock were significantly higher than those of the control group [(26.0±16.2) mmHg vs (11.0±5.6) mmHg (1 mmHg=0.133 kPa) and (4.0±1.7) mmol/L vs (1.6±0.6)mmol/L, all P=0.000]. The area under receiver operator characteristic (ROC) curve (AUC) for baseline P(c-et)CO(2) and LAC was 0.924 (95%CI: 0.851-0.996) and 0.931 (95%CI: 0.872-1.000), respectively. P(c-et)CO(2) >12.6 mmHg and LAC >2.5 mmol/L could discriminate septic shock patients from those without shock with the same sensibility of 97% and the specificity of 83% and 78% respectively. With regard to the prognosis (Day 28) of the patients with septic shock, AUC for baseline P(c-et)CO(2) and LAC was 0.709 (95%CI: 0.533-0.886) and 0.714 (95%CI: 0.545-0.883), respectively. P(c-et)CO(2) >20.0 mmHg and LAC>3.6 mmol/L could discriminate survivors from non-survivors with the same sensibility of 92% and the same specificity of 76%. All the patients in the study group completed EGDT within 6 hours after admission, 20 (57.1%) passed EGDT and 17 (85.0%) survived, 15 (42.9%) failed EGDT and 4 (26.7%) survived, and the survival rates were significantly different (F=9.844, P=0.001). After EGDT, P(c-et)CO(2) (21.0±9.5 mmHg) and LAC(3.3±2.5 mmol/L)reduced significantly compared with the baselines (P=0.008 and P=0.046), and the associated AUC was 0.905(95%CI: 0.792-1.000) and 0.747 (95%CI: 0.576-0.917)respectively. P(c-et)CO(2) > 16.5 mmHg and LAC > 3.1 mmol/L could discriminate survivors from non-survivors with the sensibility of 97% and 91%, and the specificity of 78% and 69%, respectively. Conclusions: P(c-et)CO(2) >12.6 mmHg could play the same role as LAC in recognizing early septic shock. EGDT was an effective therapy for the septic shock and P(c-et)CO(2) reflected efficacy. P(c-et)CO(2)>20 mmHg before EGDT and >16.5 mmHg after EGDT both could predict the 28 d prognosis of patients with septic shock, and the effect of the former was equal to that of LAC, but the latter was better than LAC.

Genetic differences in ChTLR15 gene polymorphism and expression involved in Salmonella enterica natural and artificial infection respectively, of Chinese native chicken breeds, with a focus on sexual dimorphism.

Chicken Toll-like receptor 15 (ChTLR15) has been shown to participate in immune activation in response to various pathogens and in the innate defence against infection. Two genetically distinct Chinese breeds of chicken (Qinyuan Partridge and Baier breeds) were used to study the correlation between ChTLR15 single nucleotide polymorphisms and the natural infection status of salmonella in hens, and also to examine genetic and sex-specific effects on ChTLR15 mRNA expression in heterophils and spleen during acute infection with Salmonella enterica serovar Enteritidis (SE) from 1 to 10 days after experimental infection. Three single-nucleotide polymorphisms (G168A, C726T and A1166G) in a single exon of ChTLR15 were identified in the two breeds, but only C726T showed a significant association with salmonella infection. Compared with layer-type Baier chicks, meat-type Qingyuan chicks showed a higher tolerance for capture stress and (SE) infection, as measured, respectively, by the modified body weight of chicks in the control group and in the infection group. Meanwhile, ChTLR15 down-regulation in heterophils and up-regulation in spleen were involved in the response to pathogenic SE colonization during the acute infection period. These significant genetic effects in females led to greater differences in both innate and adaptive immune responses than those exhibited in males. These results suggest that genetics, time and gender play important roles in the modulation of ChTLR15 mRNA level elicited by the SE-mediated immune response differentially in the two genetically distinct breeds, with a focus on sexual dimorphism.

Molecular cloning, characterization, and expression profiles of androgen receptors in spotted scat (Scatophagus argus).

Androgen plays critical roles in vertebrate reproductive systems via androgen receptors (ARs). In the present study, the full-length spotted scat (Scatophagus argus) androgen receptor (sAR) cDNA sequence was cloned from testis. The sAR cDNA measured 2448 bp in length with an open-reading frame of 2289 bp, encoding 763 amino acids. Amino acid alignment analyses showed that the sARs exhibited highly evolutionary conserved functional domains. Phylogenetically, the sARs clustered within the ARß common vertebrate group. Real-time polymerase chain reaction (RT-PCR) revealed that sAR expression varied in level and distribution throughout the tissues of both females and males. sAR expression was detected during testicular development by quantitative RT-PCR. The results showed that the highest transcription of sARs was observed in the mid-testicular stage, and remained at a high expression level until the late-testicular stage. In addition, the effects of 17α-methyltestosterone (MT) and estrogen (E2) on the expression of sARs in ovaries were determined using quantitative RT-PCR. sAR expression increased at 12 and 24 h post-MT treatment and decreased with E2 treatment. The present study provides preliminary evidence indicating gonadal plasticity of spotted scat under exogenous steroidal hormone treatments. It also provides a theoretical basis for sex reversal and production of artificial pseudo-males for female monosex breeding.

[Temperature modifies the acute effect of particulate air pollution on mortality in Jiang'an district of Wuhan].

OBJECTIVE: To analyze the temperature modification effect on acute mortality due to particulate air pollution. METHODS: Daily non-accidental mortality, cardiovascular mortality, and respiratory mortality data were obtained from Jiang'an District Center for Disease Control and Prevention. Daily meteorological data on mean temperature and relative humidity were collected from China Meteorological Data Sharing Service System. The daily concentration of particulate matter was collected from Wuhan Environmental Monitoring center. By using the stratified time-series models, we analyzed effects of particulate air pollution on mortality under different temperature zone from 2002 to 2010, meanwhile comparing the difference of age, gender and educational level, in Wuhan city of China. RESULTS: High temperature (daily average temperature > 33.4 ℃) obviously enhanced the effect of PM10 on mortality. With 10 µg/m(3) increase in PM10 concentrations, non-accidental, cardiovascular, and respiratory mortality increased 2.95% (95%CI: 1.68%-4.24%), 3.58% (95%CI: 1.72%-5.49%), and 5.07% (95%CI: 2.03%-9.51%) respectively. However, low temperature (daily average temperature <-0.21 ℃) enhanced PM10 effect on respiratory mortality with 3.31% (95% CI: 0.07%-6.64%) increase. At high temperature, PM10 had significantly stronger effect on non-accidental mortality of female aged over 65 and people with high educational level groups. With an increase of 10 µg/m(3), daily non-accidental mortality increased 4.27% (95% CI:2.45%-6.12%), 3.38% (95% CI:1.93%-4.86%) and 3.47% (95% CI:1.79%-5.18%), respectively. Whereas people with low educational level were more susceptible to low temperature. A 10 µg/m(3) increase in PM10 was associated with 2.11% (95% CI: 0.20%-4.04%) for non-accidental mortality. CONCLUSIONS: Temperature factor can modify the association between the PM10 level and cause-specific mortality. Moreover, the differences were apparent after considering the age, gender and education groups.

[Acute impact of cold spells on mortality during 2001-2011 in Jiang'an district of Wuhan, China].

OBJECTIVE: To evaluate the association between cold spells and nonaccidental mortality from 2001 to 2011 in the Jiang'an District of Wuhan, China. METHODS: We collected mortality data for December 2001 to 2010 and January to March 2002 to 2011 in the study area. According to the International Classification of Diseases, we stratified the mortality data into three cause-specific categories: nonaccidental mortality, cardiovascular mortality, and respiratory mortality. We also obtained meteorological data (from the China Meteorological Administration) and ambient pollution data (from the Wuhan Environmental Monitoring Center) during the same periods. In the present study, a cold spell was defined as 5 or more consecutive days with daily average temperatures below the 5th percentile of daily mean temperatures (2.58 ℃) from January to December in 2001-2011. A distributed lag nonlinear model (DLNM) was applied to assess the acute effect of cold spells on daily nonaccidental mortality. RESULTS: During the study period, the total number of nonaccidental deaths was 17 119, including 9 403 (75.5%) among individuals aged over 65 years; 7 968 (46.5%) people died of cardiovascular disease. According to this definition, there were a total of 13 cold spell events and 111 days of duration in Wuhan during the study period. Study days were divided into three periods: non-cold spell days, 2008 cold spell days, and cold spell days in other years. Average daily mean temperatures of the above three periods were (8.2±4.5), (-0.7±1.4), and (0.8±1.2) ℃, respectively, corresponding to average daily deaths of 14.0±4.2, 18.2±4.5, and 14.9±4.9 for nonaccidental mortality. After adjusting for long-term trends, seasonal trends, weekdays, holidays, and relative humidity, analysis by the DLNM revealed that cold spells were associated with increased mortality risk, with a cumulative relative risk (RR) of 1.56 (95% CI: 1.36-1.79) at lag 0-27 days in 2008, higher than that in other years with 1.23 (95%CI: 1.08-1.41). Cold spells were not significantly associated with respiratory mortality and people under 65 years of age; however, during the 2008 cold spell RR increased to 1.96 (95% CI: 1.62-2.37) and 1.67 (95% CI: 1.43-1.95) for cardiovascular mortality and older adults (≥65 years old), respectively; both males and females had high mortality risk, with RRs of 1.60 (95%CI:1.33-1.92) and 1.50 (95% CI: 1.23-1.84), respectively. The association between cold spells and mortality remained nearly unchanged with and without adjustment for ambient pollutants (PM10, SO2, and NO2) in the DLNMs. CONCLUSION: In Wuhan, both the 2008 cold spell and cold spells in other years were significantly associated with increased nonaccidental mortality. People with cardiovascular disease and elderly adults may be more susceptible to the impact of cold spells on mortality.

Expression of APPL1 is correlated with clinicopathologic characteristics and poor prognosis in patients with gastric cancer.

BACKGROUND: Although appl1 is overexpressed in many cancers, its status in gastric cancer (gc) is not known. In the present study, we used relevant pathologic and clinical data to investigate appl1 expression in patients with gc. METHODS: In 47 gc and 27 non-gc surgical specimens, immunohistochemistry was used to detect the expression of appl1, and reverse-transcriptase polymerase chain reaction (rt-pcr) was used to detect messenger rna (mrna). A scatterplot visualized the relationship between survival time and mrna expression in gc patients. The log-rank test and other survival statistics were used to determine the association of appl1 expression with the pathologic features of the cancer and clinical outcomes. RESULTS: In gc, appl1 was expressed in 28 of 47 specimens (59.6%), and in non-gc, it was expressed in 7 of 23 specimens (30.4%, p < 0.05). The expression of mrna in gc was 0.82 [95% confidence interval (ci): 0.78 to 0.86], and in non-gc, it was 0.73 (95% ci: 0.69 to 0.77; p < 0.05). Immunohistochemistry demonstrated that, in gc, appl1 expression was correlated with depth of infiltration (p = 0.005), lymph node metastasis (p = 0.017), and TNM stage (p = 0.022), but not with pathologic type (p = 0.41). Testing by rt-pcr demonstrated that, in gc, appl1 mrna expression was correlated with depth of infiltration (p = 0.042), lymph node metastasis (p = 0.031), and TNM stage (p = 0.04), but again, not with pathologic type (p = 0.98). The correlation coefficient between survival time and mrna expression was -0.83 (p < 0.01). Overexpression of appl1 protein (hazard ratio: 3.88; 95% ci: 1.07 to 14.09) and mrna (hazard ratio: 4.23; 95% ci: 3.09 to 15.11) was a risk factor for death in patients with gc. CONCLUSIONS: Expression of appl1 is increased in gc. Overexpression is prognostic for a lethal outcome.

Co-expression of recombinant human prolyl with human collagen α1 (III) chains in two yeast systems.

UNLABELLED: In this study, we co-expressed the human prolyl 4-hydroxylases (P4H) with human collagen α1 (III) (COL3A1) in an inducible system: Pichia pastoris (pPICZB), and one constitutive system: P. pastoris (pGAPZαB). The P4H catalyses the post-translational hydroxylation of proline residues in collagen strands. Conventional protein expression system such as bacteria and yeasts, which lack endogenous P4H, are not efficient for the production of recombinant collagen. In this study, the P4H gene was constructed in pGAPZαB plasmid and pPICZB plasmid respectively. These two plasmids were transformed in P. pastoris #1 that carrying COL3A1. Colony PCR analysis and sequencing after electroporation P. pastoris GS115 showed that the target gene had inserted successfully. The results of reverse transcript-qPCR, SDS-PAGE, Western blotting and LC-MS/MS analysis of the rhCOL3A1 demonstrated that the P4H was expressed successfully. Besides, it is noted that low copy number, constitutive system was suitable for hydroxylated rhCOL3A1. SIGNIFICANCE AND IMPACT OF THE STUDY: Successful co-expression of recombinant human collagen α1 (III) (rhCOL3A1) and human prolyl 4-hydroxylases (P4H) in Picha pastoris GS115, simultaneously results in the acquisition of rhCOL3A1 with hydroxylation of proline (Hyp). Further, this experiment also discusses that the high or low copy numbers and different promoters affect the Hyp degree of rhCOL3A1. Selecting more appropriate strains can express high degree Hyp of rhCOL3A1. This work will be helpful to the collagen structure study.

Construction of a lentiviral vector encoding heme oxygenase 1 and its introduction into mouse adipose tissue-derived stem cells.

Many studies exist concerning the use of stem cells as delivery vehicles in gene therapy, expressing genes such as vascular endothelial growth factor 165 and hepatocyte growth factor. However, few reports regarding adipose tissue-derived stem cells (ADSCs) and the heme oxygenase 1 (HO-1) gene have been published. Therefore, we established a lentiviral vector encoding HO-1 and used this to infect ADSCs with the aim of producing therapeutic seed cells. In this study, ADSCs were isolated from mouse adipose tissue (AT), cultured, and identified according to the expression of antigens on their cell surface and their capacity for multilineage differentiation. A lentiviral vector encoding HO-1 was constructed, ADSCs were infected with this, and HO-1 protein expression was examined by western blotting. Our results show that ADSCs can be isolated from mouse AT, while DNA sequencing demonstrated that HO-1 was successfully transferred to the vector fused with GFP. Following 293T cell transfection, lentivirus titers were approximately 3 x 10(8) TU/mL. Fluorescence microscopy confirmed the expression of the HO-1 construct in lentivirus-infected ADSCs and the overexpression of HO-1 protein in these cells was verified by western blot. The production of ADSCs overexpressing HO-1 described in this study may aid in the development of a novel method for the treatment of asthma.

In vivo and in vitro inhibitory action of 17ß-estradiol and environmental estrogen 4-nonylphenol on gonad-inhibiting hormone (GIH) expression in the eyestalks of Litopenaeus vannamei.

The gonad-inhibiting hormone (GIH) belongs to a neuropeptide family synthesized and released in an X-organ sinus gland complex of crustacean eyestalks. GIH inhibits crustacean ovarian maturation by suppressing vitellogenin (Vtg) synthesis, whereas estrogen is responsible for the stimulation of vitellogenesis (not established). In this study, the effects of 17ß-estradiol (E2, 10(-6) M), estrogen receptor antagonist tamoxifen (TAM, 10(-6), 10(-7), and 10(-8) M), and the environmental estrogen nonylphenol (NP, 1 µg/L and 100 µg/L) on LvGIH expression in the eyestalks of shrimp were determined by quantitative real-time PCR. Results showed that LvGIH expression decreased significantly during the L. vannamei ovarian maturation cycle. E2 and NP significantly reduced LvGIH transcripts in vivo, but TAM neutralized the inhibitory action of E2 in a dose-dependent manner (P < 0.05). In addition, the LvGIH expression levels decreased significantly in a time-dependent manner (P < 0.05) when ovary fragments were cultured in vitro with E2. The results of this study suggested that estrogen regulates GIH expression in L. vannamei eyestalks. E2 promoted ovarian development not only by directly upregulating vitellogenesis in the hepatopancreas, but it was also capable of downregulating LvGIH expression, which indirectly resulted in the stimulation of L. vannamei vitellogenesis.

Molecular characterization, expression, and functional analysis of NOD1 in Qingyuan partridge chicken.

Nucleotide-binding oligomerization domain-containing protein-1 (NOD1) is a cytoplasmic pattern recognition receptor (PRR) and a key member of the NOD-like receptor (NLR) family. It has been reported that NLRs recognize a variety of microbial infections to induce the host innate immune response via modulation of NF-κB signaling. However, no reports on chicken NOD1 have been reported to date. In the current study, the full-length cDNA sequence of NOD1 was cloned. The complete open reading frame of NOD1 contains 2856 bp and encodes a 951 amino acid protein. Structurally, it is comprised of one caspase recruitment domain at the N-terminus, seven leucine-rich repeat regions at the C-terminus, and one NACHT domain between the N and C-termini. Phylogenetic analyses showed that chicken NOD1 clusters with duck and turkey. Furthermore, tissue-specific expression analyses of chicken NOD1 were performed using quantitative reverse transcription-PCR. NOD1 is widely distributed in various tissues, with the highest expression observed in testes. Finally, induced expression of chNOD1 and its associated adaptor molecule receptor-interacting protein 2, as well as the effector molecule NF-κB, was observed following S. enterica serovar Enteritidis infection. These findings highlight the important role of chicken NOD1 in response to pathogenic invasion. The present study is the first report of the cloning, expression, and functional analysis of chicken NOD1 and provides the foundation for future research on the structure and function of chicken NOD1.

Effects of temperature and fish oil supplementation on ovarian development and foxl2 mRNA expression in spotted scat Scatophagus argus.

In this study, the complete foxl2 complementary (c)DNA sequence was isolated by simple modular-architecture research tool (SMART)er rapid amplification of cDNA ends (RACE). Two year-old female spotted scat, Scatophagus argus, were reared at different temperatures (23, 26 and 29° C) for 6 weeks, or fed with different concentrations of dietary fish oil (0, 2 or 6%) for 8 weeks. Ovarian development, serum oestradiol-17ß (E2 ) levels, as well as ovarian foxl2 expression were measured. At the end of experiment, ovarian foxl2 messenger (m)RNA expression in fish reared at 23 and 26° C was significantly higher than that in fish reared at 29° C, and that in 2 and 6% fish oil groups was also significantly higher than that in control group (P < 0·05). Serum E2 levels exhibited the same trend with foxl2 mRNA expression in temperature treatment groups and fish oil fed groups. There was a significant positive correlation between stage of oocytes and foxl2 expressions. Results showed that from 23 to 29° C, the optimal temperature for ovarian development in S. argus was 23-26° C, and 6% fish oil supplementation could effectively promote ovarian development. Optimal temperature and fish oil supplement might increase ovarian foxl2 mRNA expressions to promote ovarian development in S. argus.

Upregulation of NRAMP1 mRNA confirms its role in enhanced host immunity in post-artificial infections of Salmonella enteritidis in chicks.

1. Salmonella enteritidis (SE) is reported as the most common food-borne pathogen transmitted through poultry products. The natural resistance-associated macrophage protein 1 (NRAMP1) is a candidate gene associated with SE-mediated immune response and is related to the phagocytosis of SE. In this study, the classical single-nucleotide polymorphism (SNP) G2357A in exon 8 of the NRAMP1 gene was detected. The expression of NRAMP1 mRNA was first investigated in heterophil granulocytes and spleen in chicks from two different Chinese native breeds at 1, 3 and 10 d post-infection. In addition, the association with the effect of SE challenge was identified. 2. The G2357A SNP showed no significant association with Salmonella natural infection in birds from two different Chinese native breeds. 3. The upregulation of NRAMP1 mRNA in heterophils and spleen was involved in the response to pathogenic SE colonisation during the acute infection period in chicks. The results suggest that genetics, age, gender and interactions among these factors play important roles in the modulation of NRAMP1 mRNA expression and copy number by SE-mediated immune response in different Chinese chickens. 4. In conclusion, the enhancement of host immunity mediated by the upregulation of NRAMP1 mRNA in heterophil granulocytes and spleen might be more obvious and earlier in the chicks resistant to infections with SE than in susceptible chicks.

Molecular genetic diversity and maternal origin of Chinese black-bone chicken breeds.

Chinese black-bone chickens are valued for the medicinal properties of their meat in traditional Chinese medicine. We investigated the genetic diversity and systematic evolution of Chinese black-bone chicken breeds. We sequenced the DNA of 520 bp of the mitochondrial cyt b gene of nine Chinese black-bone chicken breeds, including Silky chicken, Jinhu black-bone chicken, Jiangshan black-bone chicken, Yugan black-bone chicken, Wumeng black-bone chicken, Muchuan black-bone chicken, Xingwen black-bone chicken, Dehua black-bone chicken, and Yanjin black-bone chicken. We found 13 haplotypes. Haplotype and nucleotide diversity of the nine black-bone chicken breeds ranged from 0 to 0.78571 and 0.00081 to 0.00399, respectively. Genetic diversity was the richest in Jinhu black-bone chickens and the lowest in Yanjin black-bone chickens. Analysis of phylogenetic trees for all birds constructed based on hyplotypes indicated that the maternal origin of black-bone chickens is predominantly from three subspecies of red jungle fowl. These results provide basic data useful for protection of black-bone chickens and help determine the origin of domestic chickens.

Genetic differ in TLR4 gene polymorphisms and expression involved in Salmonella natural and artificial infection respectively in Chinese native chicken breeds.

Salmonella enteritidis (SE) is a foodborne pathogen that negatively affects both animal and human health. Genetic variations in response to pathogenic SE colonization or to SE vaccination were measured in chicken resource populations. Toll-like receptor 4 (TLR4) is part of a group of evolutionarily conserved pattern recognition receptors involved in the activation of the immune system in response to various pathogens and in the innate defense against infection. In this study, TLR4 was investigated the association of TLR4 gene polymorphisms with Salmonella natural infection situation of birds from two distinct Chinese genetic breeds. One SNP G1894C in the second intron of chicken TLR4 (chTLR4) was scanned in the two hens breed, which showed significant association with Salmonella natural infection situation (P<0.05). Genetic variations in response to pathogenic SE colonization also existed in distinct Chinese chicken resource population. In this study, mRNA expression of TLR4 was detected to investigate the association with the effect of artificial SE challenge in heterophil granulocytes and spleen of chicks from two distinct Chinese genetic breeds at 1, 3 and 10 day post-infection during the acute infection period. It clearly showed that young chicks' response to SE infection was regulated by TLR4 mRNA expression. The results suggest that genetics, time, gender, and interactions among these factors, play important roles in TLR4 mRNA basic values and copies modulation of SE mediated immune response in distinct Chinese chickens.