ABSTRACT
Osteonecrosis of femoral head (ONFH) is one of the complications of systemic lupus erythematosus (SLE), which is characterized by complex pathogenesis and difficult treatment, and is the main cause of SLE-induced disability. Previous studies have confirmed that the JAK/STAT signaling pathway is involved in the pathological process of SLE, and the activation of JAK/STAT has also been found to be related to the occurrence of ONFH. Therefore, we speculated that the JAK/STAT signaling pathway may play an important role in the occurrence and development of SLE-ONFH. 30 female MRL/lpr mice were randomly divided into three groups: the model group (Lipopolysaccharide/24 hours, twice + Methylprednisolone/24 hours, 3 times), the control group (equal amount of PBS) and the treatment group (the model group + JAK1/2 inhibitors Baricitinib/day, 6 weeks), with 10 mice in each group. The results showed that the grip value of the model group was significantly lower than that of the control group at the 4th and 6th week (P < 0. 05), and that of the treatment group was better than that of the model group at the 6th week (P < 0. 05). At the 6th week, the mice were sacrificed to take the bilateral femoral head, and the morphology and HE staining pathological changes of the femoral head were observed. The results showed that the femoral head of the control group was spherical, transparent, hard and without cartilage defects, while that of the model group was irregular, rough, gray in color and partial defects of the femoral head. The performance of the mice in the treatment group was basically similar to that in the model group. And the overall appearance of the femoral head was more irregular than that in the control group. The color was darker than that in the control group, and there was a partial defect of the femoral head, but the degree was not as serious as that in the model group. The empty bone lacuna rate in the model group and treatment group were significantly higher than that in the control group (P < 0. 05), and the empty bone lacuna rate in the treatment group was lower than that in the model group (P < 0. 05). Western blotting, ELISA and RT-qPCR were used to detect the protein expression, phosphorylation levels, mRNA expression of the JAK/STAT pathway (JAK1, JAK2, JAK3, STAT3) in local bone tissues, and the expression of IL-6 and TNF- α in serum and local tissues. The mRNA expression of IL-6, TNF-α and STAT3 in bone tissues of the model group were significantly higher than that of the control group and treatment group, and the content of serum IL-6 and TNF- α of the model group were significantly higher than that of the treatment group and control group. The cartilage catabolic metabolites ADAMTS-4, MMP-13 and JAK/STAT pathway related proteins JAK1, p-JAK1, JAK2, p-JAK2, STAT3 and pSTAT3 in the model group were significantly higher than those in the control group and treatment group. In summary, the JAK/STAT signaling pathway is involved in the pathogenesis of ONFH in MRL/lpr lupus erythematosus mice. Selective JAK1/2 inhibitors can effectively inhibit ONFH inflammation, improve bone structures and joint functions, and may become an effective therapeutic drug for SLE ONFH.
ABSTRACT
Chengdu-Chongqing economic circle is centered on Chengdu City and Chongqing Municipality, with aims to build the “fourth growth pole” of China’s economy. During this circle, elimination of schistosomiasis had been achieved in 82.5% of the endemic counties (districts) of Sichuan Province, and schistosomiasis is not historically endemic in Chongqing Municipality; however, there is still a risk of schistosmiasis transmission in Sichuan Province and Chongqing Municipality because the natural and social factors affecting schistosomiasis transmission have not been completely eliminated in these areas. Based on the endemic situation of schistosomiasis in Sichuan Province and Chongqing Municipality, we analyzed the opportunities and challenges of schistosomiasis control during the construction of Chengdu-Chongqing economic circle, and proposed the corresponding suggestions, so as to provide insights into the sustainable development of schistosomiasis control in the context of the Chengdu-Chongqing economic circle construction.
ABSTRACT
Using silica gel column chromatography, gel chromatography and HPLC, we isolated secondary metabolites in fermentation broth of a rifamycin resistant mutation strain Streptomyces sp. HS-NF-1046R. Based on spectroscopic data, the chemical structures of three compounds were identified as 3-hydroxyl-2-N-propionyl- anthranilamide (1), 2,3-dihydro-8-hydroxy-2,2-dimethyl quinazolin-4-(1H)-one (2) and 2-aminobenzamide (3). Compounds 1 and 2, as new entities, were evaluated for cytotoxicity against A549, HepG2, HCT-116 and K562 cells using the SRB assay. Compounds 1 and 2 exhibited no cytotoxicity with IC50 over 100 μmol·L-1.
ABSTRACT
An efficient genetic transformation system and suitable promoters are essential prerequisites for gene expression studies and genetic engineering in streptomycetes. In this study, firstly, a genetic transformation system based on intergeneric conjugation was developed in Streptomyces rimosus M527, a bacterial strain which exhibits strong antagonistic activity against a broad range of plant-pathogenic fungi. Some experimental parameters involved in this procedure were optimized, including the conjugative media, ratio of donor to recipient, heat shock temperature, and incubation time of mixed culture. Under the optimal conditions, a maximal conjugation frequency of 3.05×10-5 per recipient was obtained. Subsequently, based on the above developed and optimized transformation system, the synthetic promoters SPL-21 and SPL-57, a native promoter potrB, and a constitutive promoter permE* commonly used for gene expression in streptomycetes were selected and their activity was analyzed using gusA as a reporter gene in S. rimosus M527. Among the four tested promoters, SPL-21 exhibited the strongest expression activity and gave rise to a 2.2-fold increase in β-glucuronidase (GUS) activity compared with the control promoter permE*. Promoter SPL-57 showed activity comparable to that of permE*. Promoter potrB, which showed the lowest activity, showed a 50% decrease in GUS activity compared with the control permE*. The transformation system developed in this study and the tested promotors provide a basis for the further modification of S. rimosus M527.
Subject(s)
Conjugation, Genetic , Glucuronidase/genetics , Promoter Regions, Genetic , Streptomyces rimosus/geneticsABSTRACT
otrA resembles elongation factor G (EF-G) and is considered to be an oxytetracycline (OTC)-resistance determinant in Streptomyces rimosus. In order to determine whether otrA also conferred resistance to OTC and other aminoglycosides to Streptomyces coelicolor, the otrA gene from S. rimosus M527 was cloned under the control of the strong ermE* promoter. The resulting plasmid, pIB139-otrA, was introduced into S. coelicolor M145 by intergeneric conjugation, yielding the recombinant strain S. coelicolor M145-OA. As expected S. coelicolor M145-OA exhibited higher resistance levels specifically to OTC and aminoglycosides gentamycin, hygromycin, streptomycin, and spectinomycin. However, unexpectedly, S. coelicolor M145-OA on solid medium showed an accelerated aerial mycelia formation, a precocious sporulation, and an enhanced actinorhodin (Act) production. Upon growth in 5-L fermentor, the amount of intra- and extracellular Act production was 6-fold and 2-fold higher, respectively, than that of the original strain. Consistently, reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that the transcriptional level of pathway-specific regulatory gene actII-orf4 was significantly enhanced in S. coelicolor M145-OA compared with in S. coelicolor M145.
Subject(s)
Aminoglycosides/pharmacology , Anthraquinones/metabolism , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Drug Resistance, Bacterial/genetics , Streptomyces coelicolor/metabolismABSTRACT
<p><b>Objective</b>To explore the inhibitory effect of genistein (GEN) on the proliferation of VCaP castration-resistant prostate cancer (CRPC) cells.</p><p><b>METHODS</b>VCaP CRPC cells were treated with GEN at the concentrations of 0, 12.5, 25, 50, 100, and 200 μmol/L for 24, 48, and 72 hours followed by determination of their proliferation by CCK-8 assay and their cycle by flow cytometry. The expression of Ki-67 in the cells was detected by immunocytochemistry and the levels of PSA, Cyclin D1, PCNA, and P53 determined by Western blot.</p><p><b>RESULTS</b>After 72 hours of treatment with GEN at 12.5, 25, 50, 100, and 200 μmol/L, the inhibition rates of the VCaP cells were (25.38±0.02)%, (31.14±0.29)%, (45.27±0.03)%, (52.19±0.05)%, and (68.21±0.19)%, respectively, all significantly higher than in the 0 μmol/L group (［10.08±0.02］%)(P<0.05). GEN caused the arrest of the VCaP cells in the G2/M phase (P<0.05) and inhibited the expression of Ki-67. The expressions of PSA, Cyclin D1, and PCNA were gradually down-regulated while that of P53 up-regulated with the increased concentration of GEN (P<0.05).</p><p><b>CONCLUSIONS</b>GEN inhibits the proliferation of VCaP CRPC cells by arresting the cell cycle with related protein expression changes.</p>
ABSTRACT
Preoperative detection of lymph nodes (LNs) metastasis is always highly challenging for radiologists nowadays. The utility of quantitative dynamic contrast-enhanced magnetic resonance imaging (QDCE-MRI) in identifying LNs metastasis is not well understood. In the present study, 59 patients with histologically proven rectal carcinoma underwent preoperative QDCE-MRI. The short axis diameter ratio, long axis diameter ratio, short-to-long axis diameter ratio and QDEC-MRI parameters (K(trans), Kep, fPV and Ve) values were compared between the non-metastatic (n=44) and metastatic (n=35) LNs groups based on pathological examination. Compared with the non-metastatic group, the metastatic group exhibited significantly higher short axis diameter (7.558±0.668 mm vs. 5.427±0.285 mm), K(trans) (0.483±0.198 min(-1) vs. 0.218±0.116 min(-1)) and Ve (0.399±0.118 vs. 0.203±0.096) values (all P<0.05). The short-to-long axis diameter ratio, long axis diameter ratio, Kep and fPV values did not show significant differences between the two groups. In conclusion, our results showed that for LNs larger than 5 mm in rectal cancer, there are distinctive differences in the K(trans) and Ve values between the metastatic and non-metastatic LNs, suggesting that QDCE-MRI may be potentially helpful in identifying LNs status.
Subject(s)
Female , Humans , Male , Contrast Media , Therapeutic Uses , Diagnosis, Differential , Lymph Nodes , Diagnostic Imaging , Pathology , Lymphatic Metastasis , Diagnosis , Diagnostic Imaging , Pathology , Magnetic Resonance Imaging , Rectal Neoplasms , Diagnosis , Diagnostic Imaging , PathologyABSTRACT
To verify the effect of echinacoside on replication and antigen expression of hepatitis B virus (HBV) by using HBV-transfected HepG2. 2. 15 cells as the in vitro model. The ELISA method was used to determine HBeAg and HBsAg levels in cellular supernatants. The effect of echinacoside on HBV replication was studied by using HBV transgenic mice as the in vivo model. First of all, the HBV DNA level in hepatic tissues was quantified with PCR method. Meanwhile, the serum transaminase levels and hepatic pathological changes were also evaluated. Subsequently, HBV transgenic mice were divided into five groups: the control group, the lamivudine group (50 mg · kg(-1)) and echinacoside high, medium and low dose group (50, 25 and 12.5 mg · kg(-1)). The mice were orally administered with drugs once per day for 30 days. At the 31st day, the mice serum was separated to measure HBsAg, HBeAg and HBV DNA. Additionally, the liver HBV DNA level and histopathological change were detected. The results indicated that echinacoside at 50 and 100 mg · L(-1) suppressed significantly HBsAg and HBeAg expressions on the sixth day, with the maximum inhibition ratios of 42.68% and 46.29%; And echinacoside at 100 mg · L(-1) also showed an inhibitory effect on HBV DNA. Besides, echinacoside at 50 mg · kg(-1) inhibited significantly HBsAg and HBeAg expressions of HBV transgenic mice, with the inhibition ratios of 42.82% and 29.12%, and reduced markedly the serum HBV DNA level in HBV transgenic mice. In conclusion, the study suggested that echinacoside has a strong effect against HBV replication and antigen expression.
Subject(s)
Animals , Female , Humans , Male , Mice , DNA, Viral , Blood , Glycosides , Pharmacology , Hep G2 Cells , Hepatitis B Surface Antigens , Blood , Hepatitis B e Antigens , Blood , Hepatitis B virus , Physiology , Mice, Inbred C57BL , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>To study the effect of aluminume adjuvant and immunization schedule on immunogenicity of Sabin inactivated poliovirus vaccine.</p><p><b>METHODS</b>Four batches of Sabin IPV were produced by different concentrations of type 1, 2, and 3 poliovirus and administrated on three-dose schedule at 0, 1, 2 months and 0, 2, 4 months on rats. Serum samples were collected one month after each dose and neutralizing antibody titers against three types poliovirus were determined by micro-neutralization assay.</p><p><b>RESULTS</b>The GMTs of neutralizing antibodies against three types poliovirus increased significantly and the seropositivity rates were 100% in all groups after 3 doses. There was no significant difference between two immunization schedules, and the 0, 2, 4 month schedule could induce higher level neutralizing antibody compared to the 0, 1, 2 month schedule. The groups with aluminum adjuvant could induce higher level neutralizing antibody compared to the groups without adjuvant.</p><p><b>CONCLUSION</b>Aluminum djuvant and immunization schedule could improve the immunogenicity of Sabin IPV.</p>
Subject(s)
Animals , Female , Male , Rats , Adjuvants, Immunologic , Pharmacology , Aluminum Hydroxide , Pharmacology , Antibodies, Viral , Blood , Immunization Schedule , Poliovirus Vaccine, Oral , Allergy and Immunology , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To explore the feasibility of karyotype analysis using cells cultured from fetal bladder centesis samples.</p><p><b>METHODS</b>Samples were derived from fetal bladder centesis for 3 fetuses featuring giant bladder and oligohydramnios. Following in vitro culture, cells were routinely processed and stained for chromosome analysis.</p><p><b>RESULTS</b>For all 3 cases, cell culture has achieved success. Sufficient metaphase cells were obtained for chromosome counting and karyotype analysis. The karyotypes of the 3 fetuses were respectively 46, XY, 46, XX, t(1;5)(q22;q12)[7]/46, XX[4], and 46, XY.</p><p><b>CONCLUSION</b>Cells cultured from fetal bladder centesis may be used for karyotype analysis following in vitro culturing. This new approach can enable prenatal chromosome analysis for fetuses featuring smaller gestational weeks, giant bladder and oligohydramnios.</p>
Subject(s)
Female , Humans , Pregnancy , Cells, Cultured , Karyotyping , Prenatal Diagnosis , Methods , Urinary Bladder , Congenital AbnormalitiesABSTRACT
<p><b>OBJECTIVE</b>To study the associations between paraoxonase, 55 Met/Leu (PON1 55 Met/ Leu), paraoxonase2 148 Ala/Gly (PON2 148 Ala/Gly) and manganese superoxide dismutase 9 Ala/Val (MnSOD 9 Ala/Val) genetic polymorphisms and coronary heart disease (CHD), plasma activities of paraoxonase (PON), total superoxide dismutase (T-SOD), MnSOD, as well as plasma concentration of maleic dialdehyde (MDA).</p><p><b>METHODS</b>Using PCR-RFLP method to identify genotype of PON1 55 Met/Leu, PON2 148 Ala/Gly and MnSOD 9 Ala/Val genetic polymorphisms, and using colorimetry to detect plasma activities of PON, T-SOD, MnSOD and plasma concentration of MDA in 262 CHD patients and 100 controls.</p><p><b>RESULTS</b>Compared with controls, the plasma activities of PON [(349.27 +/- 138.36 vs. 454.75 +/- 166.00) nmol x min(-1) x ml(-1), P < 0.001], T-SOD [(23.61 +/- 16.51 vs. 44.01 +/- 22.68) U/ml, P < 0.001] and MnSOD [(21.56 +/- 13.11 vs. 28.79 +/- 8.65) U/ml, P < 0.001] reduced obviously,while plasma MDA concentration increased markedly [(2.47 +/- 0.73 vs. 2.15 +/- 0.55)nmol/ml, P < 0.01] in CHD patients. There were more LM genotype and Met allele of PON, 55 Met/Leu (24.8% vs. 1.4%, P < 0.001 and 12.4% vs. 0.5%, P = 0.001, respectively), GG and AG genotype and G allele of PON2 148 Ala/Gly (11.8% vs. 5.0%, P < 0.001, 48.1% vs. 24.0%, P < 0.001 and 36.0% vs. 17.0%, P < 0.001, respectively) and AA genotype, A allele of MnSOD 9 Ala/Val genetic polymorphisms (64.2% vs. 43.0%, P = 0.001 and 80.0% vs. 67.0%, P = 0.014, respectively) in CHD patients than in controls. The activities of plasma PON and T-SOD were lower in individuals with PON1 55 LM genotype than those with LL genotype. The activity of plasma PON was also lower in individuals with PON2 148 GG/AG genotype than those with AA genotype. The activities of plasma PON and MnSOD depressed in individuals with MnSOD AA genotype compared with those with VV genotype. Logistic regression analysis demonstrated that PON1 55 LM genotype, PON2 148 GG/AG genotype and G allele were independent risk factors for CHD.</p><p><b>CONCLUSION</b>The antioxidative ability decreased, while lipid superoxide increased in CHD patients. Gene polymorphisms of PON1 55 Met/Leu, PON2 148 Ala/Gly and MnSOD 9 Ala/Val seemed to involve in the morbidity of CHD by influencing the plasma activities of PON and MnSOD.</p>
Subject(s)
Humans , Aryldialkylphosphatase , Genetics , Metabolism , Case-Control Studies , China , Coronary Disease , Genetics , Genotype , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment Length , Superoxide Dismutase , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the relationships between paraoxonase 1 55 Met/Leu (PON1 55Met/Leu), paraoxonase 2 148 Ala/Gly(PON2 148Ala/Gly) genetic polymorphisms and coronary artery disease(CAD), plasma activities of paraoxonase (PON), total superoxide dismutase (T-SOD), as well as plasma concentration of maleic dialdehyde (MDA).</p><p><b>METHODS</b>The PCR-RFLP method was applied to identify the genetic polymorphisms of PON1 55Met/Leu and PON2 148Ala/Gly, and the colorimetry way was used to detect plasma activities of PON, T-SOD and plasma MDA concentration of 262 CAD patients and 100 controls.</p><p><b>RESULTS</b>Comparing with control, the CAD patient had the obviously lower activities of enzymes PON (349.27+/- 138.36 nmol/min.mL vs 454.75+/- 166.00 nmol/min.mL, P< 0.001) and T-SOD (23.61+/- 16.51 U/mL vs 44.01+/- 22.68 U/mL, P< 0.001) while getting the plasma MDA concentration increased markedly(2.47+/- 0.73 nmol/mL vs2.15+/- 0.55 nmol/mL, P< 0.01). The CAD patient had more LM genotype and M allele of PON1 55Met/Leu(24.8% vs 1.4%, P< 0.001 and 12.4% vs 0.5%, P was 0.001 respectively), GG and AG genotype and G allele of PON2 148 Ala/Gly(11.8% vs 5.0%, P< 0.001; 48.1% vs 24.0%, P< 0.001 and 36.0% vs 17.0%, P< 0.001 respectively) than control did. The activities of plasma PON and T-SOD were lower in individuals with PON??1 55 LM genotype than those with LL genotype(304.73+/- 125.04 vs 394.84+/- 154.87 nmol/min.mL and 24.89+/- 16.14 vs 30.22+/- 21.29 U/mL, P< 0.001 and P< 0.05 respectively). The activity of plasma PON was also lower in individuals with PON2 148 GG/AG genotype than that with AA genotype(281.47+/- 84.70 vs 356.00+/- 145.95 vs 417.34+/- 159.00 nmol/min.mL, P< 0.001). Logistic regression analysis showed that PON1 55 LM genotype (OR 29.08, 95%CI 2.88-294.04, P was 0.004) and M allele(OR 15.17, 95%CI 1.32-174.29, P was 0.029), PON2 148 GG/AG genotype (OR 2.32, 95%CI 1.52-3.54, P< 0. 001) and G allele (OR 3.24, 95%CI 1.38-7.61, P was 0.007) were independent risk factors for CAD.</p><p><b>CONCLUSION</b>The CAD patient has the obviously low activities of plasma PON and T-SOD but on the contrary, get the plasma MDA concentration increased markedly. PON1 55 LM genotype and M allele, PON2 148 GG/AG genotype and G allele are the risk factors for coronary artery disease, and the activity of plasma PON is also markedly reduced in individuals with above genotypes.</p>