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The relationship between antibiotic resistance and bacterial virulence has not yet been fully explored. Here, we use Edwardsiella tarda as the research model to investigate the proteomic change upon oxytetracycline resistance (LTB4-R OTC ). Compared to oxytetracycline-sensitive E. tarda (LTB4-S), LTB4-R OTC has 234 differentially expressed proteins, of which the abundance of 84 proteins is downregulated and 15 proteins are enriched to the Type III secretion system, Type VI secretion system, and flagellum pathways. Functional analysis confirms virulent phenotypes, including autoaggregation, biofilm formation, hemolysis, swimming, and swarming, are impaired in LTB4-R OTC . Furthermore, the in vivo bacterial challenge in both tilapia and zebrafish infection models suggests that the virulence of LTB4-R OTC is attenuated. Analysis of immune gene expression shows that LTB4-R OTC induces a stronger immune response in the spleen but a weaker response in the head kidney than that induced by LTB4-S, suggesting it's a potential vaccine candidate. Zebrafish and tilapia were challenged with a sublethal dose of LTB4-R OTC as a live vaccine followed by LTB4-S challenge. The relative percentage of survival of zebrafish is 60% and that of tilapia is 75% after vaccination. Thus, our study suggests that bacteria that acquire antibiotic resistance may attenuate virulence, which can be explored as a potential live vaccine to tackle bacterial infection in aquaculture.

Background: The epithelial-mesenchymal transition (EMT) of human bronchial epithelial cells (HBECs) is essential for airway remodeling during asthma. Wnt5a has been implicated in various lung diseases, while its role in the EMT of HBECs during asthma is yet to be determined. This study sought to define whether Wnt5a initiated EMT, leading to airway remodeling through the induction of autophagy in HBECs., Methods: Microarray analysis was used to investigate the expression change of WNT5A in asthma patients. In parallel, EMT models were induced using 16HBE cells by exposing them to house dust mites (HDM) or interleukin-4 (IL-4), and then the expression of Wnt5a was observed. Using in vitro gain- and loss-of-function approaches via Wnt5a mimic peptide FOXY5 and Wnt5a inhibitor BOX5, the alterations in the expression of the epithelial marker E-cadherin and the mesenchymal marker protein were observed. Mechanistically, the Ca 2+ /CaMKII signaling pathway and autophagy were evaluated. An autophagy inhibitor 3-MA was used to examine Wnt5a in the regulation of autophagy during EMT. Furthermore, we used a CaMKII inhibitor KN-93 to determine whether Wnt5a induced autophagy overactivation and EMT via the Ca 2+ /CaMKII signaling pathway., Results: Asthma patients exhibited a significant increase in the gene expression of WNT5A compared to the healthy control. Upon HDM and IL-4 treatments, we observed that Wnt5a gene and protein expression levels were significantly increased in 16HBE cells. Interestingly, Wnt5a mimic peptide FOXY5 significantly inhibited E-cadherin and upregulated α-SMA, Collagen I, and autophagy marker proteins (Beclin1 and LC3-II). Rhodamine-phalloidin staining showed that FOXY5 resulted in a rearrangement of the cytoskeleton and an increase in the quantity of stress fibers in 16HBE cells. Importantly, blocking Wnt5a with BOX5 significantly inhibited autophagy and EMT induced by IL-4 in 16HBE cells. Mechanistically, autophagy inhibitor 3-MA and CaMKII inhibitor KN-93 reduced the EMT of 16HBE cells caused by FOXY5, as well as the increase in stress fibers, cell adhesion, and autophagy., Conclusion: This study illustrates a new link in the Wnt5a-Ca 2+ /CaMKII-autophagy axis to triggering airway remodeling. Our findings may provide novel strategies for the treatment of EMT-related diseases., (© 2024. The Author(s).)

Objectives: To explore the association between assisted reproductive technology (ART) and birth weight discordance in twins (BWDT)., Methods: A retrospective analysis was conducted on twin infants born between January 2011 and December 2020 at the Third Affiliated Hospital of Guangzhou Medical University, with complete basic birth data. The impact of ART on the occurrence of BWDT was identified by the multivariate logistic regression analysis., Results: A total of 3 974 pairs of twins were included, with 1 431 conceived naturally and 2 543 through ART. Neonates in the ART group had higher birth weights than those in the naturally conceived group ( P <0.001). The incidence of BWDT was lower in the ART group compared to the naturally conceived group (16.17% vs 21.09%, P <0.001). The multivariate logistic regression analysis, adjusting for confounding factors such as maternal age, parity, pre-pregnancy body mass index, gestational diabetes, hypothyroidism, gestational age, and chorionic properties, showed no significant difference in the risk of BWDT between the ART and naturally conceived groups ( P >0.05)., Conclusions: ART is not associated with the risk of BWDT.

Multidrug-resistant Edwardsiella tarda threatens both sustainable aquaculture and human health, but the control measure is still lacking. In this study, we adopted functional proteomics to investigate the molecular mechanism underlying norfloxacin (NOR) resistance in E. tarda . We found that E. tarda had a global proteomic shift upon acquisition of NOR resistance, featured with increased expression of siderophore biosynthesis and Fe 3+ -hydroxamate transport. Thus, either inhibition of siderophore biosynthesis with salicyl-AMS or treatment with another antibiotic, kitasamycin (Kit), which was uptake through Fe 3+ -hydroxamate transport, enhanced NOR killing of NOR-resistant E. tarda both in vivo and in vitro . Moreover, the combination of NOR, salicyl-AMS, and Kit had the highest efficacy in promoting the killing effects of NOR than any drug alone. Such synergistic effect not only confirmed in vitro and in vivo bacterial killing assays but also applicable to other clinic E. tarda isolates. Thus, our data suggest a proteomic-based approach to identify potential targets to enhance antibiotic killing and propose an alternative way to control infection of multidrug-resistant E. tarda .

We previously found that the disorder of soluble epoxide hydrolase (sEH)/cyclooxygenase-2 (COX-2)-mediated arachidonic acid (ARA) metabolism contributes to the pathogenesis of the non-alcoholic fatty liver disease (NAFLD) in mice. However, the exact mechanism has not been elucidated. Accumulating evidence points to the essential role of cellular senescence in NAFLD. Herein, we investigated whether restoring the balance of sEH/COX-2-mediated ARA metabolism attenuated NAFLD via hepatocyte senescence. A promised dual inhibitor of sEH and COX-2, PTUPB, was used in our study to restore the balance of sEH/COX-2-mediated ARA metabolism. In vivo , NAFLD was induced by a high-fat diet (HFD) using C57BL/6J mice. In vitro , mouse hepatocytes (AML12) and mouse hepatic astrocytes (JS1) were used to investigate the effects of PTUPB on palmitic acid (PA)-induced hepatocyte senescence and its mechanism. PTUPB alleviated liver injury, decreased collagen and lipid accumulation, restored glucose tolerance, and reduced hepatic triglyceride levels in HFD-induced NAFLD mice. Importantly, PTUPB significantly reduced the expression of liver senescence-related molecules p16, p53, and p21 in HFD mice. In vitro , the protein levels of γH2AX, p53, p21, COX-2, and sEH were increased in AML12 hepatocytes treated with PA, while Ki67 and PCNA were significantly decreased. PTUPB decreased the lipid content, the number of β-gal positive cells, and the expression of p53, p21, and γH2AX proteins in AML12 cells. Meanwhile, PTUPB reduced the activation of hepatic astrocytes JS1 by slowing the senescence of AML12 cells in a co-culture system. It was further observed that PTUPB enhanced the ratio of autophagy-related protein LC3II/I in AML12 cells, up-regulated the expression of Fundc1 protein, reduced p62 protein, and suppressed hepatocyte senescence. In addition, PTUPB enhanced hepatocyte autophagy by inhibiting the PI3K/AKT/mTOR pathway through Sirt1, contributing to the suppression of senescence. PTUPB inhibits the PI3K/AKT/mTOR pathway through Sirt1, improves autophagy, slows down the senescence of hepatocytes, and alleviates NAFLD.

Necroptosis, a recently described form of programmed cell death, is the main way of alveolar epithelial cells (AECs) death in acute lung injury (ALI). While the mechanism of how to trigger necroptosis in AECs during ALI has been rarely evaluated. Long optic atrophy protein 1 (L-OPA1) is a crucial mitochondrial inner membrane fusion protein, and its deficiency impairs mitochondrial function. This study aimed to investigate the role of L-OPA1 deficiency-mediated mitochondrial dysfunction in AECs necroptosis. We comprehensively investigated the detailed contribution and molecular mechanism of L-OPA1 deficiency in AECs necroptosis by inhibiting or activating L-OPA1. First, our data showed that L-OPA1 expression was downregulated in the lungs and AECs under the lipopolysaccharide (LPS) challenge. Furthermore, inhibition of L-OPA1 aggravated the pathological injury, inflammatory response, and necroptosis in the lungs of LPS-induced ALI mice. In vitro, inhibition of L-OPA1 induced necroptosis of AECs, while activation of L-OPA1 alleviated necroptosis of AECs under the LPS challenge. Mechanistically, inhibition of L-OPA1 aggravated necroptosis of AECs by inducing mitochondrial fragmentation and reducing mitochondrial membrane potential. While activation of L-OPA1 had the opposite effects. In summary, these findings indicate for the first time that L-OPA1 deficiency mediates mitochondrial fragmentation, induces necroptosis of AECs, and exacerbates ALI in mice., (© 2022 Wiley Periodicals LLC.)

Background: Fibrodysplasia ossificans progressiva (FOP) is a rare and disabling heritable connective tissue disease that is difficult to treat. This study seeks to explore the clinical characteristics, clinical manifestations, treatment and prognosis of FOP to provide a clinical basis for its early diagnosis and treatment., Methods: Twenty-six children with FOP were retrospectively analyzed in terms of their onset, clinical manifestations, auxiliary examinations and treatment., Results: Among the 26 cases, the youngest age of manifestation of mass was 8 days after birth, and the average age was 3 years and 2 months. The peak age was 2-5 years old. Inflammatory mass and toe-finger deformity are the main early clinical manifestations of the disease. These inflammatory masses often lead to hard osteogenic deposits that initially mainly involve the central axis, such as the neck (22/26, 84.6%), back (20/26, 76.9%), and head (13/26, 50%). Toe-finger deformity mainly manifests as symmetrical great toe deformity, or short and deformed thumb and little finger. The diagnosis of FOP requires typical clinical manifestations or ACVR1 gene detection. The main therapeutic drugs for FOP include glucocorticoids and non-steroidal anti-inflammatory drugs. Although not compliant with the recommended medical management of FOP, in our clinical practice children with uncontrollable illness could be treated using a variety of immunosuppressive agents in combination., Conclusions: FOP is a rare autosomal dominant heritable disease. The main clinical manifestations observed in this study were recurrent inflammatory mass and toe-finger deformity. If the diagnosis and treatment are not performed in a timely manner, serious complications are likely to affect the prognosis. Therefore, early diagnosis and active treatment should be performed.

Objective: To determine the presence of vasculogenic mimicry (VM) and expression of Sphingosine kinase 1 (SphK1) and Connexin43 (Cx43) in colorectal cancer (CRC) tissues, and to identify their inter-relationships and associations with multiple pathologic parameters., Methods: Ninety-two CRC specimens and normal pericarcinoma tissues were analyzed for expression of SphK1 and Cx43 using immunohistochemistry, and for identification of VM using CD34-periodic acid-Schiff dual staining., Results: The positive rate of SphK1 expression was greater in CRC cells than pericarcinoma cells (85.87% vs. 33.70%, P < 0.05). In contrast, the positive rate of Cx43 expression was greater in pericarcinoma cells than in CRC cells (58.70% vs. 92.39%, P < 0.05). Analysis of CRC tissues indicated that expression of SphK1 was associated with poor differentiation, advanced tumor stage, lymph node metastasis, and the presence of VM ( P < 0.05 for each comparison). Expression of Cx43 was associated with high differentiation and the presence of VM ( P < 0.05 for each comparison). Patient sex, age, tumor size, depth of invasion, and distant metastasis were unrelated to the expression of either protein. There was a significant correlation between the expression of SphK1 and Cx43 ( P < 0.05). Analysis of overall patient survival indicated that SphK1 positivity and the presence of VM were significantly associated with poor survival, but Cx43 positivity had no relationship with survival., Conclusion: SphK1 protein expression was significantly greater in CRC tissues than pericarcinoma tissues, suggesting this protein may be associated with the pathogenesis of CRC. In addition, the significant correlation between expression of SphK1 and Cx43 in CRC tissues suggests their interaction may impact the pathogenesis of CRC., Competing Interests: None., (IJCEP Copyright © 2018.)

Lung diseases in neonates can be life-threatening condition and may result in respiratory failure and death. Chest X-ray is a traditional diagnostic technique that results in radiation exposure to patients. Lung ultrasound is a user-friendly imaging technique that has been increasingly used in clinical practice in recent years and presents the advantages of real-time imaging and without radiation. Here we review the sonographic appearances of common neonatal lung diseases and present demonstration of typical cases., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest to declare.

Ischaemia-reperfusion injury (I/RI) is a common cause of acute kidney injury (AKI). The molecular basis underlying I/RI-induced renal pathogenesis and measures to prevent or reverse this pathologic process remains to be resolved. Basic fibroblast growth factor (FGF2) is reported to have protective roles of myocardial infarction as well as in several other I/R related disorders. Herein we present evidence that FGF2 exhibits robust protective effect against renal histological and functional damages in a rat I/RI model. FGF2 treatment greatly alleviated I/R-induced acute renal dysfunction and largely blunted I/R-induced elevation in serum creatinine and blood urea nitrogen, and also the number of TUNEL-positive tubular cells in the kidney. Mechanistically, FGF2 substantially ameliorated renal I/RI by mitigating several mitochondria damaging parameters including pro-apoptotic alteration of Bcl2/Bax expression, caspase-3 activation, loss of mitochondrial membrane potential and K ATP channel integrity. Of note, the protective effect of FGF2 was significantly compromised by the K ATP channel blocker 5-HD. Interestingly, I/RI alone resulted in mild activation of FGFR, whereas FGF2 treatment led to more robust receptor activation. More significantly, post-I/RI administration of FGF2 also exhibited robust protection against I/RI by reducing cell apoptosis, inhibiting the release of damage-associated molecular pattern molecule HMBG1 and activation of its downstream inflammatory cytokines such as IL-1α, IL-6 and TNF α. Taken together, our data suggest that FGF2 offers effective protection against I/RI and improves animal survival by attenuating mitochondrial damage and HMGB1-mediated inflammatory response. Therefore, FGF2 has the potential to be used for the prevention and treatment of I/RI-induced AKI., (© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

Objective: To analyze the epidemiological and etiological characteristics of diarrheal disease among children under 5 years of age in Guangdong province, in 2012., Methods: 64 hospitals in 21 cities were chosen as the diarrheal syndromic surveillance sites, of which 14 hospitals were selected to carry out etiological surveillance among children under 5 years of age, including isolation and culture of both Vibrio cholera and Shigella as well as nucleic acid detection of rotavirus and norovirus by PCR. Descriptive method was used to analyze data from syndromic and etiological surveillance programs on diarrheal, from 1932 parents of the children., Results: In 2012, the outpatient attendance rate on diarrheal among children under 5 years was 0.8%. The proportion of diarrheal in children under 5-year-olds was 63.5%, among the total number of diarrheal outpatients at the outpatient clinics under surveillance program. The morbidity of infectious diarrhea was 1454.5/10 million in children under 5 years of age. A total number of 1932 specimens were collected from children under 5 years of age, in the outpatient department. Among these specimens,Vibrio cholera appeared all negative but one was Shigella positive and proved to be Sh. sonnei. The positive rates of rotavirus and norovirus were 14.1% (273/1932)and 16.9% (326/1932). Both rotavirus and norovirus were found in 24 specimens, with a positive rate as 1.2% . 112 specimens were successfully gene sequenced for rotavirus, of which 33.9% as G1[P8] genotype, 25.9% as G9[P8], 12.5% as G2[P4] and 9.8% as G3[P8] respectively. 90 specimens were successfully gene-sequenced for norovirus, of which 76.7% as G II.4 genotype. Genetic subtypes of G II. 4/2006b, accounted for 50.0% and could be detected around the year except for June and December. New G II. 4/Sydney Strain&#95;2012 was first detected in August and became the predominant in December. In addition, 5 specimens belonged to G I genotype with other 16 subtypes of G II., Conclusion: Results from our study proved that children under 5 years of age belonged to high-risk group for diarrheal disease in Guangdong province. Rotavirus and norovirus were both diverse in terms of genome.

Objective: To explore the correlation of serum ferritin (SF) and systemic onset juvenile idiopathic arthritis (SOJIA) so as to determine the prognostic values of SF for SOJIA., Methods: All samples were collected from 92 juvenile idiopathic arthritis (JIA) patients at Beijing Children's Hospital between February 2005 to September 2012. Their age range was 2-15 years. There were macrophage activation syndrome (MAS, n = 25), polyarticular JIA (n = 33) and oligoarticular JIA (n = 30). And 47 healthy children and another 30 with acute infective diseases were selected as control groups respectively. Blood samples were collected and SF was measured in different disease phases.Other parameters include leucocyte, hemoglobin, platelet, C-reactive protein and erythrocyte sedimentation rate.Statistics of SF level at different groups as well as at different disease phases were performed., Results: The SF level of active phase SOJIA patients was significantly higher (P < 0.01) than that in Other groups. And its level in the active phase of SOJIA was significantly higher than that in patients during the recovery phase. The SF level in patients with MAS was significantly higher than that in those without MAS., Conclusion: Correlated with the course of SOJIA, the level of SF may judge the disease activity and predict the outcomes of SOJIA.

Objective: To study the effects of post-discharge formula (PDF) for preterm infants, breast milk (BM) and term infant formula (TF) on increase rates of body weight, length and head circumference in preterm and low-birth-weight infants (PLBWIs) from discharge to 3 months after birth, and to provide a reference for the choice of feeding pattern for PLBWIs., Methods: A total of 407 PLBWIs discharged from the newborn departments of ten hospitals in Guangzhou City and Foshan City in Guangdong Province, China were chosen for this study. According to feeding pattern, they were assigned to three groups: PDF-fed (n=258), BM-fed (n=58) and TF-fed (n=91). Their body weight, length and head circumference were measured at 3 months after birth, and the increase rates of growth indices relative to baseline values (at birth) were calculated and compared., Results: At 3 months after birth, the PDF-fed group had significantly greater body weight, length and head circumference than the BM-fed and TF-fed groups (P<0.05). The increase rates of body weight and length were significantly higher in the PDF-fed group than in the BM-fed and TF-fed groups (P<0.05)., Conclusions: Compared with those fed with BM and TF after discharge, the PDF-fed PLBWIs have higher increase rates of body weight and length and show greater body weight and length at 3 months after birth. However, further study is needed to investigate the long-term effects.

Objective: This study aims to establish and evaluate the methodology of isolated rabbit eye (IRE) test., Methods: IRE test was performed according to modifications of the in vitro toxicology (INVITTOX) Protocol No.85: Rabbit enucleated eye test by European Centre for the Validation of Alternative Methods (ECVAM), and then 26 chemicals and 26 cosmetic products were tested in both in vitro IRE and in vivo Draize tests. A statistical analysis was conducted to determine the relevance of the IRE test to the data generated in the Draize test., Results: IRE test was established successfully in our laboratory. It was shown that ranking correlation and class concordance were fairly well between the IRE test and the Draize test for 26 reference chemicals (Fisher's Exact Test χ(2)=51.314, P<0.001; McNemar P=0.261; Gamma=0.960, P<0.001; Kappa=0.843, P<0.001) and 26 cosmetic products (Fisher's Exact Test χ(2)=15.522, P<0.001; McNemar P=0.311; Gamma=0.967, P<0.001; Kappa=0.611, P<0.001)., Conclusion: IRE test was established successfully for in vitro testing of eye irritation as an alternative to Draize test., (Copyright © 2012 The Editorial Board of Biomedical and Environmental Sciences. Published by Elsevier B.V. All rights reserved.)

Cytokine-induced killer (CIK) cell therapy, an adoptive T-cell immunotherapy, has been reported to be a safe and effective mode of treatment for patients with metastatic diseases, lymphoma and acute leukaemia. To investigate the clinical efficacy of cytokine-induced killer cells for the treatment of refractory lymphoma, the present clinical study was conducted. A total of 8 male patients with a mean age of 41 years (range 22-65) who were pathologically diagnosed with malignant lymphoma (Hodgkin's disease, 2 and non-Hodgkin's lymphoma, 6) were enrolled. CIK cells were expanded by priming with IFN-γ, monoclonal antibody (mAb) to CD3 and IL-1α, followed by the addition of IL-2 the following day using peripheral blood mononuclear cells (PBMCs) of the 8 male patients. The CIK cells were then transfused back to the patients as treatment. On day 13, the CIK cell count reached 7-18×10(19) (mean, 12.7×10(9)), a 44- to 140-fold increase (mean, 98-fold). The average percentage of cells expressing CD3(+), CD4(+), CD8(+) and CD3(+)CD56(+) were also increased from 50.9±3.5, 29.9±1.7, 41.3±3.2, 1.6±0.2% to 90.2±1.6, 40.6±5.5, 52.8±4.9 and 33.1±4.0%, respectively. Patients showed measurable radiographic tumor reduction, increased T-cell subset levels, and relief of symptoms after treatment. No severe toxicity or side effects were reported. CIK cells developed by this culture method have a high in vitro proliferation rate and tumor-killing capacity. In conclusion, CIK cell treatment of patients with malignant lymphoma achieves effective clinical responses, causing few side effects.

Objective: To investigate the possibility of directional differentiation of human adipose stem cells (hADSCs) into endothelial cells (EC), so as to provide seed cells for tissue engineered vessels., Methods: hADSCs were isolated from human adipose tissue by collagenase digestion, cultured and amplified by adherence to flasks. Then hADSCs were directionally induced to differentiate into EC by a combination of fibronectin (FN), endothelial cells support liquid (EGM2-MV) containing various growth factors and high concentration of VEGF165 (50 ng/ml). Then, the cells morphology, phenotype and function were identified., Results: Highly homologous hADSCs were obtained, and then hADSCs were directionally differentiated into EC. CD31 and CD34, the specific markers for EC, and vascular endothelial growth factor receptor (KDR) were positive by immunohistochemical staining and RT-PCR. In addition, unique Weibel-Palade bodies in EC were observed under transmission electron microscope. Functionally, hADSCs could swallow Dil-Ac-LDL and form tube-like structures in matrigel after endothelial differentiation., Conclusions: hADSCs can be successfully induced to differentiate into endothelial cells in vitro.

Objective: To explore the toxicity of joint exposure to diazinon, propoxur and bisphenol A on phagocytosis., Methods: Flow cytometer was employed to detect the influence of diazinon and bisphenol A, propoxur and bisphenol A in mixture (mixed according to ratio of IC(50)) on mouse macrophage RAW264.7 cells' function to phagocyte fluorescent microspheres, adopting the percentage of phagocytic cells (PP) and the phagocytic index (PI) as measurement indicators. The final concentrations of mixture of diazinon and bisphenol A were (0.4 + 0.1), (3.6 + 0.7), (36.2 + 7.2), (43.4 + 8.7), (52.1 + 10.4), (62.5 + 12.5), (75.0 + 15.0) µg/ml; while those of mixture of propoxur and bisphenol A were (0.2 + 2.0 × 10(-2)), (2.4 + 0.2), (23.7 + 2.0), (35.6 + 3.0), (53.3 + 4.4), (80.0 + 6.7), (120.0 + 10.0) µg/ml. Then based on the dose-response relationship, a 2 × 2 factorial design was then carried out among different doses of mixture with statistical significance to statistically evaluate the interaction between diazinon and bisphenol A, propoxur and bisphenol A., Results: After the joint exposure, compared to the control group (PP = (23.6 ± 2.2)%; PI = 0.36 ± 0.03), any dose of the mixture of diazinon and bisphenol A ((52.1 + 10.4), (62.5 + 12.5), (75.0 + 15.0) µg/ml) could significantly increase the levels of PP ((29.0 ± 1.4)%, t = 3.89, P < 0.05; (30.2 ± 2.3)%, t = 4.74, P < 0.05; (35.0 ± 3.4)%, t = 8.21, P < 0.05) and PI (0.43 ± 0.03, t = 3.86, P < 0.05; 0.41 ± 0.02, t = 2.95, P < 0.05; 0.46 ± 0.03, t = 5.34, P < 0.05); while that of propoxur and bisphenol A ((35.6 + 3.0), (53.3 + 4.4), (80.0 + 6.7), (120.0 + 10.0) µg/ml) reduced the levels of PP ((20.6 ± 1.1)%, t = -3.00, P < 0.05; (20.2 ± 1.0)%, t = -3.42, P < 0.05; (19.4 ± 1.3)%, t = -4.23, P < 0.05; (18.8 ± 2.1)%, t = -4.81, P < 0.05) and PI (0.31 ± 0.01, t = -4.75, P < 0.05; 0.31 ± 0.01, t = -4.58, P < 0.05; 0.30 ± 0.01, t = -4.92, P < 0.05; 0.27 ± 0.02, t = -7.80, P < 0.05) on the contrary. The 2 × 2 factorial design was carried out between the mixture of diazinon (60.0 µg/ml; PP = (28.5 ± 3.4)%; PI = 0.49 ± 0.07) and bisphenol A (12.0 µg/ml; PP = (35.7 ± 2.7)%; PI = 0.67 ± 0.07), and the mixture of propoxur (48.0 µg/ml ; PP = (28.1 ± 2.2)%; PI = 0.48 ± 0.04) and bisphenol A (4.0 µg/ml; PP = (34.4 ± 2.7)%; PI = 0.59 ± 0.07). The mixture of diazinon and bisphenol A (PP = (30.4 ± 1.4)%, F(interaction) = 6.22, P < 0.05; PI = 0.53 ± 0.03, F(interaction) = 7.35, P < 0.05) and the mixture of propoxur and bisphenol A (PP = (27.5 ± 4.1)%, F(interaction) = 4.56, P < 0.05; PI = 0.46 ± 0.08, F(interaction) = 11.13, P < 0.05) both showed a significant antagonistic interaction on phagocytosis of RAW264.7 cell., Conclusion: It is suggested that the interactions between diazinon & bisphenol A and propoxur & bisphenol A both played the antagonistic role on phagocytic function of macrophages in vitro.

Background: Genetic variants of the genes encoding HIV-1 co-receptors and their ligands, CCR5-Delta32, CCR5m303A, CCR2-64I and SDF1-3'A, are implicated in the susceptibility to HIV-1 infection, and the prevalence of these mutations varies by ethnicity. However, little is known about their distribution in Uighurs., Objectives: This study aimed at characterizing the frequency of these HIV-related gene variants in a high-risk Uighur population., Study Designs: A total of 251 HIV-1 seropositive and 238 seronegative high-risk Uighurs were recruited and their genotypes of CCR5-Delta32, CCR5m303A, CCR2-64I and SDF1-3'A were analyzed by PCR and PCR-ligase detection reaction (PCR-LDR)., Results: The allelic frequency of CCR5-Delta32, CCR5m303A, CCR2-64I and SDF1-3'A was 4.40%, 2.66%, 25.66% and 57.36%, respectively, in this population. Apparently, the Uighur population has low frequency of CCR5-Delta32 and CCR5m303A, but high frequency of CCR2-64I and SDF1-3'A. While there was no significant difference in the frequency of CCR5-Delta32, CCR2-64I and SDF1-3' A between HIV-1 seropositive and seronegative groups the frequency of CCR5m303A in HIV-1 seropositive group was significantly higher than that in seronegative group (P=0.006, OR=3.982 and 95%CI 1.514-10.476)., Conclusions: Our data suggest that the CCR5-Delta32, CCR2-64I and SDF1-3'A variants may have limited effect on protecting from HIV-1 infection in Uighurs. Rather, the CCR5m303A may be associated with the risk for HIV-1 infection in high-risk Uighurs.

Objective: To research the co-infections of HIV and human herpesvirus 8 (HHV8) in Uygur high-risk groups of HIV infection in a city Xinjiang., Methods: All 468 Uygurs at high HIV risk registered in the sentinel monitoring system in 2006 were enrolled in this study. The antibodies to HHV8 latency-associated nuclear antigens 1 (LANA1), lytic antigens open reading frame 65 (ORF65) and K8.1 were measured by enzyme linked immunosorbent assay (ELISA). Chi-square test and non-condition Logistic regression model were used for data analysis., Results: Of 468 sera samples, 67 (14.3%) were HIV and HHV8 co-infection positive.Male's HIV and HHV8 co-infection rate (22.6%, 54/239) was higher than the female's (5.7%, 13/229) (chi(2) = 27.285, P < 0.001). For those above 24 year old, HIV and HHV8 co-infection rate (15.8%, 65/412) was higher than the < 24 year old group's (3.6%, 2/56) (chi(2) = 5.987, P = 0.014). The group of Elementary school and illiterate people's HIV and HHV8 co-infection rate (20.7%, 40/193) was higher than the junior middle school and the above culture (9.8%, 27/275) (chi(2) = 10.999, P = 0.001). For the unmarried people, the co-infecting rate of HIV and HHV8 for the married, the cohabitants, the divorced or the widowers were 16.9% (14/83), 12.2% (42/345), 27.5% (11/40) respectively. There was significantly statistical difference among three marital status (chi(2) = 7.399, P = 0.025). Injecting drug users' HIV and HHV8 co-infection rate (26.5%, 50/189) was higher than non-injecting drug users' (6.1%, 17/279) (chi(2) = 38.083, P < 0.001), and stratified by gender, OR(M-H) was 4.207 (95%CI: 1.529 - 11.578). Via non-condition logistic stepwise regression analysis, only injecting drug use entered model, compared with non-injecting drug users, injecting drug users were more dangerous for HIV and HHV8 co-infecting (OR = 5.544; 95%CI: 3.081 - 9.975)., Conclusion: The HIV and HHV8 co-infection rate was higher in the Uygurs at high HIV risk in Xinjiang. Injecting drug use is a risk factor of the HIV and HHV8 co-infection, which might be one of routes of HIV and HHV8 co-infection among this group.

Aim: Oncolytic adenovirus, also called conditionally replicating adenovirus (CRAD), can selectively propagate in tumor cells and cause cell lysis. The released viral progeny can infect neighboring cancer cells, initiating a cascade that can lead to the ultimate destruction of the tumor. Suicide gene therapy using herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir (GCV) offers a potential treatment strategy for cancer and is undergoing preclinical trials for a variety of tumors. We hypothesized that HSV-TK gene therapy combined with oncolytic adenoviral therapy would have an enhanced effect compared with the individual effects of the therapies and is a potential novel therapeutic strategy to treat liver cancer., Methods: To address our hypothesis, a novel CRAD was created, which consisted of a telomerase-dependent oncolytic adenovirus engineered to express E1A and HSV-TK genes (Ad-ETK). The combined effect of Ad-ETK and GCV was assessed both in vitro and in vivo in nude mice bearing HepG2 cell-derived tumors. Expression of the therapeutic genes by the transduced tumor cells was analyzed by RT-PCR and Western blotting., Results: We confirmed that Ad-ETK had antitumorigenic effects on human hepatocellular carcinoma (HCC) both in vitro and in vivo, and the TK/GCV system enhanced oncolytic adenoviral therapy. We confirmed that both E1A and HSV-TK genes were expressed in vivo., Conclusion: The Ad-ETK construct should provide a relatively safe and selective approach to killing cancer cells and should be investigated as an adjuvant therapy for hepatocellular carcinoma.

Objective: To investigate the effect of adenovirus vector encoding human vascular endothelial growth factor receptor-2 (hVEGFR-2 or hKDR) on breaking the immune tolerance and inducing immunity against murine hepatocellular carcinomas., Methods: Human and mouse KDR cDNA were cloned from human umbilical vein endothelial cells (HUVEC) and C57BL/6 mouse embryo cells respectively using RT-PCR, and then Ad hKDR and Ad mKDR were constructed. Seven days after immunization of the mice with Ad hKDR or Ad mKDR, an analysis of cytotoxic activity of antigen-specific cytotoxic T lymphocytes (CTL) was made by lactate dehydrogenase (LDH) release assay, in which splenocytes of the immunized mice acted as effectors and Hepa 1-6/mKDR cells as the targets. In addition, the survival of the mice immunized with Hepa 1-6 hepatoma cells was checked., Results: Seven days after immunization, the 6 h killing activities of CTL elicited by the Ad hKDR were 84.3%+/-6.7%, 71.5%+/-5.2%, and 44.6%+/-4.7% at the ratio of the effectors:targets (E:T) of 100:1, 50:1, and 25:1, respectively. Correspondingly, the CTL activities by Ad mKDR were 65.2%+/-6.1%, 46.7%+/-5.0%, and 22.6%+/-3.7%. Sixty percent of the Ad hKDR-immunized mice with 5*10(6) Hepa 1-6 hepatoma cells were still alive two months after the inoculation, whereas just 40% of the Ad mKDR-immunized mice with 2*10(6) Hepa 1-6 cells survived two months. When CD8+ or CD4+ T lymphocytes were deleted in the mice the above mentioned CTL activities and protection of the mice from tumors disappeared., Conclusion: Adenovirus vector-mediated xenogeneic KDR can effectively break the immune tolerance to hepatocellular carcinomas in an animal model and induce a strong antigen-specific T cell response, which is dependent on CD8+ and CD4+ T cells.

Background & Objective: Primary hepatocellular carcinoma (HCC) is a common malignant tumor. Up to date, no effective treatment for HCC is available. This study was to investigate the synergic effect of adenoviral vector-encoding xenogeneic alpha-fetoprotein (AFP) and interferon-gamma (IFN-gamma) on the immunity against HCC in mice., Methods: IFN-gamma gene was cloned using reverse transcription-polymerase chain reaction (RT-PCR). Replication-defective adenovirus encoding both human AFP and murine IFN-gamma was constructed. The cytotoxic activity of antigen-specific cytotoxic T lymphocytes (CTLs) was detected 7 days after the intradermal immunization of C57BL/6 mice with Ad-hAFP, Ad-IFN-gamma or Ad-hAFP/IFN-gamma using standard (51)Cr release assay. The survival of mice after the subcutaneous inoculation of 5x10(6) hepatoma Hepa 1-6 cells was checked., Results: The cytotoxic activity of CTLs elicited by Ad-hAFP/IFN-gamma was much stronger than that by Ad-hAFP or Ad-IFN-gamma alone. At an effector:target (E:T) ratio of 10:1, the cytotoxic activities of CTLs in Ad-hAFP/IFN-gamma, Ad-hAFP, and Ad-IFN-gamma groups were (43.8+/-5.5)%, (28.2+/-3.2)%, and (12.8+/-1.9)%, at a ratio of 30:1, were (79.6+/-6.4)%, (51.9+/-4.3)%, and (15.6+/-2.3)%, and at a ratio of 90:1, were (88.2+/-6.3)%, (62.5+/-4.8)%, and (26.5+/-2.4)%, respectively. The tumor formation rates were 80% in Ad-hAFP-immunized mice and 100% in Ad-IFN-gamma-immunized mice at 2 months after inoculation with Hepa1-6 cells; no tumor formed in Ad-hAFP/IFN-gamma-immunized mice, and all mice in this group survived during two-month observation., Conclusion: Ad-hAFP can efficiently induce immunity against HCC in mice, and IFN-gamma enhances such an effect.

Objective: To establish the 3T3 mouse fibroblast neutral red uptake (NRU-PT) phototoxicity test method, and evaluate the practicality of the method in detecting potential phototoxicity of the cosmetic products., Methods: Fifteen phototoxic and 9 non-phototoxic chemicals were tested in our laboratories, the phototoxic potential of the test chemicals was evaluated in a prediction model in which either the photo irritation factor (PIF) or the mean photo effect (MPE) was compared with the coherence and sensitivity of the method. 20 kinds of functional cosmetics were detected and the results were analyzed by the 3T3 NRU-PT in vitro and Guinea pig skin phototoxicity test (in vivo)., Results: Both PIF and MPE of the chemicals were highly reproduced, and the correlation between in vitro and in vivo data was almost perfect. All the non-phototoxic provided a negative result, while 14 of the 15 phototoxic tested chemicals gave clear positive results. For cosmetics, the correlation between in vitro and in vivo data was consistent., Conclusion: The 3T3 NRU PT test was established successfully, it should be used as a good alternative method for assessing the phototoxic potential of the chemicals and cosmetics in China.

Objective: To develop a more economic and applicable substitute for modification of dendritic cells (DCs) by capped mRNA in induction of anti-tumor immunity., Methods: Four DNA plasmids as templates of mRNA in vitro transcription were constructed: pmRNA luciferase (Luc), pmRNA internal ribosomal entry site (IRES)-Luc, pmRNA ovalbumin (OVA), and pmRNA IRES-OVA. The translational efficiency of uncapped-Luc, capped-Luc, or IRES-Luc mRNA in murine DCs was detected via a Luc reporter system. Nine C57BL/6 mice were divided into 3 equal groups to be injected intraperitoneally with DCs transfected with IRES-OVA or capped-OVA mRNA and untreated DCs respectively, 1 weeks later the mice were injected with splenocytes wrapped with the target peptide or control peptide labeled by CFSE, and 4 hours later the mice were killed and suspension of splenocytes was made to test the activity of cytotoxic lymphocytes (CTLs). Another 30 mice were divided into 3 equal groups to undergo immunization by DCs as mentioned above, 1 week later mice melanoma cells of the line MO5 stably expressing OVA were injected the caudal vein, and 3 weeks later the mice were killed and their lungs were taken out to observe the metastasis of tumor, using OVA as a target antigen., Results: Insertion of IRES into upstream of gene of interest in mRNA transcriptional templates didn't affect the yield of mRNA in vitro transcription. The level of Luc activity expressed by IRES-Luc mRNA in the DCs was 20 times higher than that by expressed by Uncapped Luc mRNA, and one time higher than that by Capped-Luc mRNA 8 h after transfection of DC. Expression of Luc could be detected up to 96 h after the transfection with IRES-Luc and Capped-Luc mRNA. Four hours after the injection of peptides the CTL activity of the mice immunized with DCs pulsed with Capped-OVA mRNA was (28 +/- 3)%, not significantly different from that of the mice immunized with DCs pulsed with IRES-OVA mRNA [(32 +/- 4)%, P > 0.05]. Mata static melanoma nodules could be seen in all control mice, only one mouse of the Capped-OVA mRNA group, and none of the mice of the IRES-OVA mRNA group., Conclusion: IRES-containing tumor-associated antigen (TAA) mRNA as a more economic and applicable substitute fully replaces Capped-TAA mRNA for mRNA-based DC vaccines and DCs pulsed with IRES-containing TAA mRNA induces the same effective antigen-specific cellular response as the DCs pulsed with Capped-TAA mRNA.

Objective: To explore a flow cytometry (FCM)-based method for discriminating aneugen- or clastogen-induced micronuclei., Methods: Cells were stained with anti-CD71-FITC and PI, and the PI fluorescent signal intensity of micronucleated reticulocyte (MN-RET) in the peripheral blood of NIH mouse treated with COL or CP was detected by flow cytometry., Results: The ratio of the median of the intensity of MN-RET fluorescent signals to that of nucleated cell was low in the cyclophosphamide treated mouse, while the median was high in the colchicine treated mouse., Conclusion: The flow cytometry-based micronucleus assay can be used to discriminate primarily smaller MN induced by the clastogen exposure from the larger MN induced by an aneugen.

Objective: To investigate the enhanced effect of bone marrow-derived dendritic cells (DC) pulsed with SVYDFFVWL, a MHC class I peptide located in 180-188 amino acid residues of human melanoma-associated antigen tyrosinase- related protein 2 ( hTRP2) on the immunity against melanomas elicited by adenovirus encoding hTRP2 (Ad hTRP2)., Methods: The mice were intradermally immunized with Ad hTRP2, and three weeks later with Ad hTRP2 or DC/SVYDFFVWL once more. Analysis of CTL killing activity and IFN-gamma-producing CD8 + T cells in the total CD8 + T cells of spleen were made using in vivo CTL and intracellular staining of IFN-gamma, respectively. Additionally, the survival of mice was checked after the subcutaneous inoculation with mouse melanoma B16. F10 cells., Results: The 6 h CTL killing and IFN-gamma producing CD8 +T cells in the total CD8 ' T cells of spleens were 68. 40%+/-5. 50% and 0. 67%+/-0.16% in Ad hTRP2 (priming)-Ad hTRP2 (boosting) group,28. 50%+/-6.40% and 0.22%+/-0.07% in DC/SVYDFFVWL (priming)-DC/ SVYDFFVWL (boosting) group,and 98. 90%+/-0.90% and 1.05%+/-0.21% in Ad hTRP2 (priming)-DC/ SVYDFFVWI, (boosting) group, respectively. In the tumor-bearing model, none of mice survived in DC/SVYDFFVWL (priming)-DC/SVYDFFVWL (boosting) group, and just only 40% of mice were tumor-free in Ad hTRP2 (priming) -Ad hTRP2 (boosting) group, whereas 100% of mice survived in Ad hTRP2 (priming)-DC/SVYDFFVWL (boosting) group., Conclusion: Boosting with DC/ SVYDFFVWL can significantly enhance the immunity against melanomas elicited by priming with Ad hTRP2, indicating that first priming with Ad hTRP2 and then boosting with DC/SVYDFFVWL is a potentially effective regimen for overcoming the disadvantage that anti-tumor immune response can not be significantly increased by readministration of adenovirus.

Objective: To investigate the effects of dendritic cells (DCs) infected with adenovirus vector encoding xenogeneic alpha-fetoprotein (AFP) on breaking the immune tolerance and induction of immunity against hepatocellular carcinomas., Methods: Human and mouse alpha-fetoprotein full-length cDNA were cloned from human HepG2 and mouse Hepa 1 - 6 hepatoma cell lines, respectively, using RT-PCR, and then inserted into adenoviral shuttle vectors to construct Ad hAFP and Ad mAFP. Mice were immunized with Ad hAFP-infected DC and in vitro CTL activity against Hepa 1 - 6 cells was examined by standard (51)Cr release assay. Survival was studied of the immunized mice, with or without depletion of CD8+ or CD4+ T cells, inoculated with Hepa 1 - 6 mouse hepatoma cells., Results: The lytic activity of CTL elicited by the Ad hAFP-infected DCs were much stronger than that by Ad mAFP-infected DCs. 80% of the Ad hAFP/DCs-immunized mice of the inoculated with 5 x 10(6) Hepa 1 - 6 hepatoma cells were still alive two months after inoculation. However, the Ad mAFP/DCs-immunized mice inoculated with 1 x 10(6) Hepa 1 - 6 cells were just 20% surviving two months later. Depletion of CD8+ or CD4+ T cells abolished such an antigen-specific immunity elicited by the DCs infected with Ad hAFP., Conclusion: Adenovirus vector-mediated xenogeneic AFP-infected DCs can effectively break the immune tolerance to hepatocellular carcinomas in an animal model and induce strong antigen-specific T cell response, which are dependent on CD8+ and CD4+ T cells.

Objective: To analyse related risk factors of classic Kaposi' s sarcoma in Xinjiang., Methods: A 1:4 case-control study was used and the conditional logistic regression model was performed in this study. Cases were followed up in Xinjiang while controls were selected by the same sex, nation and age within 5 years with cases., Results: Interleukin-6,vascular endothelial growth factor, beta-MG, neopterin, human herpevirus 8, were found to be associated with Kaposi's sarcoma risk in one-way variance model while beta2 -MG and human herpevirus 8 entered the multiple conditional logistic regression model, and their ORs were 1.002(95%CI: 1.000-1.003), 81.041 (95%CI: 1.790-3669.620)., Conclusion: There was a correlate relationship between beta2 -MG and classic Kaposi's sarcoma being found that human herpevirus 8 exposure related factors seemed to have played important roles on classic Kaposi's sarcoma in Xinjiang.

Objective: To investigate the relationship between anti-tumor immune response and autoimmunity elicited by adenovirus-mediated xenogeneic melanoma-related antigen tyrosinase-related protein 2 (TRP2)., Methods: Mice divided into three groups were intradermally immunized by dl 70-3 (control group), Ad hTRP2, and Ad hTRP2 respectively. Analysis of the killing activity and production of interferon (IFN) gamma of tumor-specific cytotoxia T lymphocytes (CTL) from the C57BL/6 mice (3 mice per group) immunized with adenoviral vector encoding human or murine TRP2 (Ad hTRP2 or Ad mTRP2) were made using in vivo CTL and intracellular staining of IFN-gamma respectively. Additionally, the survival of mice (10 mice per group) immunized with Ad hTRP2 or Ad mTRP2 was checked after the subcutaneous inoculation with mouse melanoma B16.F10 cells., Results: The CTL lysis activity elicited by Ad hTRP2 was related to the duration of immunization, which increased up to peak one week after the immunization and subsequently decreased. Moreover, the Ad hTRP2-elicited CTL lysis was dose-dependent. The 6 h CTL killing and spleen IFN-gamma-producing CD8(+) T cells in total CD8(+) T cells by one week's immunization with Ad hTRP2 were 94% +/- 5% and 0.87% +/- 0.12%, respectively, whereas the 6 h CTL killing and spleen IFN-gamma-producing CD8(+) T cells by Ad mTRP2 were 22% +/- 8% and 0.15% +/- 0.05% respectively. On the other hand, 80%of the mice inoculated with 1 x 10(6) B16.F10 cells a week after the immunization with Ad hTRP2 were tumor-free for three months, but companied by formation of vitiligo in the Ad injection sites. However, only 20% of the Ad mTRP2-immunized mice inoculated with 1 x 10(5) B16.F10 cells survived, and no vitiligo was formatted., Conclusion: Adenovirus-mediated xenogeneic melanoma-related antigen TRP2 effectively breaks immune tolerance, thus inducing strong antigen-specific cytotoxic T cell response, simultaneously companied by obvious autoimmune injury.

Objective: An human telomerase reverse transcriptase (hTERT) promoter was cloned to investigate the effect of simplex virus-thymidine kinase (HSV-tk) gene-ganciclovir (GCV) system under control of this promoter on lung cancer cells A549., Methods: Specific primers were designed to amplify the core hTERT promoter from HepG2 genome. A set of expression vectors encoding LacZ or tk gene under control of hTERT or hCMV promoter was constructed through standard molecular cloning methods. After the transfection with lipofectamine 2000, reverse transcriptional PCR (RT-PCR) and beta-Gal staining were performed to examine the activity of the hTERT promoter. The cytotoxic effects of GCV/tk on A549 and MRC5 cells transfected with the plasmids encoding tk gene were evaluated by MTT assay., Results: The hTERT promoter containing -208-+40 bp of the upstream sequence of human telomerase was successfully cloned. beta-Gal staining and RT-PCR were used to detect the expression of Lac Z and the transcription of tk gene under control of the hTERT promoter, respectively, in A549 rather than MRC5 cells. Cyototoxic effect of GCV was observed only in the A549 but not MRC5 cells after the transfection with pDC511 hTERT/tk, and the effect was dose-dependent. The effect, however, was observed in cells transfected with pDC518 hCMV/tk., Conclusion: The hTERT promoter cloned in this study can specifically control the target gene expression in telomerase-positive tumor cells but not the normal cells, suggesting that the HSV-tk/GCV system under control of the hTERT promoter is a promising targeted gene therapy for malignant tumors.