ABSTRACT
The acaricide bifenazate acts as complex III inhibitor whereas cyenopyrafen and SYP-9625 act as complex II inhibitors. All these acaricides are commonly used to control two-spotted spider mite (TSSM), Tetranychus urticae Koch. We examined field-evolved and laboratory-selected resistance of TSSM to these three acaricides and determined cross-resistance among them. Six field populations of TSSM showed low levels of resistance to bifenazate with resistance ratios ranging from 2.20 to 10.65 compared to a susceptible strain. SYP-9625, structurally similar to cyenopyrafen, showed slightly higher activity to TSSMs but significant cross-resistance in both field populations and a laboratory-selected strain by SYP-9625. However, low levels of resistance to these two chemicals were found in field populations even when used for short time periods. Cross-resistance was not found between bifenazate and Complex II inhibitors, cyenopyrafen and SYP-9625, in both field populations and the laboratory-selected strain. Field-evolved resistance of TSSM to the tested acaricides is still low and should be delayed by the implementation of resistance management practices. Cross-resistance between cyenopyrafen and SYP-9625 is obvious, so they should not be used together in resistance management strategies based on mode of action rotation.
Subject(s)
Acaricides/pharmacology , Biological Evolution , Drug Resistance , Tetranychidae/drug effects , Acrylonitrile/analogs & derivatives , Acrylonitrile/pharmacology , Animals , Carbamates/pharmacology , China , Hydrazines/pharmacology , Pyrazoles/pharmacology , Tetranychidae/physiologyABSTRACT
Chemical control is important in the management of the tobacco whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). Susceptibility of B. tabaci to insecticides may vary among different developmental stages and geographical populations. In this study, we examined toxicity of seven commonly-used insecticides to B. tabaci MED in four field populations from China. Avermectin has high level of toxicity to all stages of B. tabaci MED in all four populations. Cyantraniliprole and sulfoxaflor have high toxicity to adults. Spirotetramat, cyantraniliprole and flonicamid have high toxicity to nymphs but not adults. Acetamiprid, cyantraniliprole and sulfoxaflor have high toxicity to eggs. However, the relative toxicity of B. tabaci MED to these chemicals varied across different populations, with little consistency in population differences across developmental stages. Our findings together with some instances where LC95 values were higher than field recommended dosages indicate field-evolved resistance to insecticides (such as thiamethoxam and sulfoxaflor) and stage-specific mechanisms that will influence effective control of B. tabaci MED by insecticides.
Subject(s)
Hemiptera/drug effects , Insecticides/pharmacology , Animals , China , Hemiptera/growth & development , Nymph/drug effects , Nymph/growth & developmentABSTRACT
To develop a new control method for the two-spotted spider mite (TSSM), Tetranychus urticae, we investigated the effect of controlled atmospheres of carbon dioxide (CO2) on TSSM mortality under different concentrations and treatment periods, and evaluated the impact of treatments on seedlings of five host plants of TSSM. Egg hatching rate of TSSM was reduced to 37.7, 5.4 or 0% after 24 h treatment involving concentrations of 16.7, 33.3 or 50%, respectively. Mobile stages (nymphs and adult) of TSSM were completely controlled after 24 h treatment at concentrations higher than 33.3%. After 4 h at concentrations of 33.3 or 50%, 1st-day survival rate for all mobile stages was 45.3 or 36.0%, respectively, whereas after 8 or 16 h treatments, all values were decreased to zero. Seedlings of four major host plants of TSSM (cucumber, eggplant, rape, green peppers) were damaged to varying degrees after 24 h at the three concentrations, but strawberry, another host plant, was not damaged. Cucumber suffered the most serious damage, resulting in wilting and death. In conclusion, controlled atmospheres of CO2 can kill TSSM, particularly at high concentrations and with long treatment times. It can be used to control TSSM on strawberry, but should be used cautiously on other host plants.
Subject(s)
Acaricides , Carbon Dioxide , Crops, Agricultural , Tetranychidae , Tick Control , Animals , Crops, Agricultural/growth & development , Seedlings/growth & developmentABSTRACT
This study assessed the clinical application of transvaginal three-dimensional ultrasound (3D TVUS) in the diagnosis of congenital uterine malformation. A retrospective study was performed on 62 patients with congenital uterine malformation confirmed hysteroscopically and/or laparoscopically. The patients were subjected to transvaginal two-dimensional ultrasound (2D TVUS) and 3D TVUS. The accuracy rate was compared between the two methods. The accuracy rate of 3D TVUS was (98.38%, 61/62), higher than that of 2D TVUS (80.65%, 50/62). 3D TVUS coronal plane imaging could demonstrate the internal shape of the endometrial cavity and the external contour of the uterine fundus. It allowed accurate measurement on the coronary plane, and could three-dimensionally show the image of cervical tube, thereby providing information for the diagnosis of some complex uterine malformation. 3D TVUS imaging can obtain comprehensive information of the uterus malformation, and it is superior to 2D TVUS for the diagnosis of congenital uterine malformations, especially complex uterine anomaly.
Subject(s)
Imaging, Three-Dimensional/methods , Ultrasonography/methods , Urogenital Abnormalities/diagnostic imaging , Uterus/diagnostic imaging , Adult , Female , Humans , Reproducibility of Results , Retrospective Studies , Sensitivity and Specificity , Urogenital Abnormalities/diagnosis , Uterus/abnormalities , Young AdultABSTRACT
Knee osteoarthritis has become one of the common diseases of the elderly, total knee arthroplasty ï¼TKAï¼ is the most effective treatment for end-stage knee osteoarthritis at present. In TKA, the effective restoration of the lower extremity alignment is one of the key factors for the success of the operation, which greatly affects the postoperative clinical effect and prosthesis survival rate of patients. Mechanical alignment is a TKA alignment method which is first proposed, recognized and widely used in TKA. In recent years, with the in-depth research on the lower limb alignment and the rapid development of computer technology, the alignment technology in TKA has realized the transformation from "unified" to "individualized", two-dimensional to three-dimensional. New alignment methods, such as adjusted mechanical alignment, anatomic alignment, kinematic alignment, inverse kinematic alignment, restricted kinematic alignment and functional alignment have been proposed to provide surgeons with more choices. However, there is no conclusion on which alignment method is the best choice. This paper summarizes the current research status, advantages and disadvantages of various alignment methods in TKA, and aims to provide some reference for the selection of alignment methods in TKA.
Subject(s)
Arthroplasty, Replacement, Knee , Knee Prosthesis , Osteoarthritis, Knee , Humans , Aged , Arthroplasty, Replacement, Knee/adverse effects , Osteoarthritis, Knee/surgery , Knee Joint/surgery , Lower Extremity/surgery , Biomechanical PhenomenaABSTRACT
The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is one of the most important pests that has developed high pesticide resistance. The resistances of five Chinese populations of this moth, four resistant strains (from Beijing, Henan, Fujian, and Guangdong) and one susceptible strain, to five pesticides were determined, and the activities of carboxylesterase, glutathione S-transferase, and acetylcholine esterase were tested in all five populations. The correlations between pesticide resistance and enzyme activity were analyzed. The results showed that the resistance status to the five pesticides was different among the five populations. The resistance ratios of the Beijing and Henan populations to spinosad were 5.84 and 8.22, respectively, and those to beta-cypermethrin were 4.91 and 4.98, respectively. These ratios were higher than those for the Fujian and Guangdong populations. The Fujian population was more sensitive to abamectin and chlorpyrifos than the susceptible population (the resistance ratios were 0.14 and 0.91, respectively); in fact, the median lethal concentration for P. xylostella was significantly higher for chlorpyrifos than that for any of the other four pesticides. The carboxylesterase activity in P. xylostella showed positive correlations with the resistance to spinosad, beta-cypermethrin, chlorpyrifos, and abamectin, but no correlation was observed between the carboxylesterase activity and resistance to emamectin benzoate, between glutathione S-transferase activity and resistance to any of the five pesticides tested, or between acetylcholine esterase activity and any of the pesticides except for emamectin benzoate.
Subject(s)
Moths/enzymology , Acetylcholinesterase/genetics , Acetylcholinesterase/metabolism , Animals , Carboxylesterase/genetics , Carboxylesterase/metabolism , Glutathione Transferase/genetics , Glutathione Transferase/metabolism , Insect Proteins/genetics , Insect Proteins/metabolism , Insecticide Resistance , Larva/drug effects , Larva/enzymology , Larva/genetics , Moths/drug effects , Moths/genetics , Moths/growth & developmentABSTRACT
Polydnaviruses (PDVs), obligatory symbionts with parasitoid wasps, function as host immune suppressors and growth and development regulator. PDVs can induce host haemocyte apoptosis, but the underlying mechanism remains largely unknown. Here, we provided evidence that, during the early stages of parasitism, the activated Cotesia vestalis bracovirus (CvBV) reduced the overall number of host haemocytes by inducing apoptosis. We found that one haemocyte-highly expressed CvBV gene, CvBV-26-4, could induce haemocyte apoptosis. Further analyses showed that CvBV-26-4 has four homologs from other Cotesia bracoviruses and BV from wasps in the genus Glyptapanteles, and all four of them possessed a similar structure containing 3 copies of a well-conserved motif (Gly-Tyr-Pro-Tyr, GYPY). Mass spectrometry analysis revealed that CvBV-26-4 was secreted into plasma by haemocytes and then degraded into peptides that induced the apoptosis of haemocytes. Moreover, ectopic expression of CvBV-26-4 caused fly haemocyte apoptosis and increased the susceptibility of flies to bacteria. Based on this research, a new family of bracovirus genes, Bracovirus apoptosis-inducing proteins (BAPs), was proposed. Furthermore, it was discovered that the development of wasp larvae was affected when the function of CvBV BAP was obstructed in the parasitized hosts. The results of our study indicate that the BAP gene family from the bracoviruses group is crucial for immunosuppression during the early stages of parasitism.
Subject(s)
Moths , Polydnaviridae , Wasps , Animals , Polydnaviridae/genetics , Hemocytes , Larva , ApoptosisABSTRACT
OBJECTIVE: To compare survival outcomes between primary radical surgery and primary radiation in early cervical cancer. METHODS: Patient information was extracted from the Surveillance, Epidemiology, and Results database. Patients diagnosed with early cervical cancer of stage T1a, T1b, and T2a (American Joint Committee on Cancer, 7th edition) from 1998 to 2015 were included in this study after propensity score matching. Overall survival (OS) was analyzed using the Kaplan-Meier method. RESULTS: Among the 4964 patients included in the study, 1080 patients were identified as having positive lymph nodes (N1), and 3884 patients were identified as having negative lymph nodes (N0). Patients with primary surgery had significantly longer 5-year OS than those with primary radiotherapy in both the N1 group (P<0.001) and N0 group (P<0.001). In the subgroup analysis, similar results were found in patients with positive lymph nodes of stage T1a (100.0% vs. 61.1%), T1b (84.1% vs. 64.3%), and T2a (74.4% vs. 63.8%). In patients with T1b1 and T2a1, primary surgery resulted in longer OS than primary radiation, but not in patients with T1b2 and T2a2. In multivariate analysis, the primary treatment was identified as an independent prognostic factor in both N1 and N0 patients (HRN1=2.522, 95% CI=1.919-3.054, PN1<0.001; HRN0=1.895, 95% CI=1.689-2.126, PN0<0.001). CONCLUSION: In early cervical cancer stage T1a, T1b1, and T2a1, primary surgery may result in longer OS than primary radiation for patients with and without lymph node metastasis.
Subject(s)
Lymph Nodes , Uterine Cervical Neoplasms , Female , Humans , Lymph Nodes/surgery , Lymph Nodes/pathology , Lymphatic Metastasis/pathology , Neoplasm Staging , Uterine Cervical Neoplasms/radiotherapy , Uterine Cervical Neoplasms/surgery , Uterine Cervical Neoplasms/pathologyABSTRACT
OBJECTIVE: The present study was designed to evaluate the effects of adjuvant chemotherapy (CT) vs. radiotherapy (RT, alone or combined with CT) on the prognosis of patients with high-risk, early-stage (stage I and stage II) endometrioid endometrial carcinoma. METHODS: This single-center retrospective clinical study was conducted in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between 2010 and 2019. In the present study, endometrioid endometrial carcinoma patients, who underwent total hysterectomy and bilateral salpingo-oophorectomy followed by postoperative adjuvant CT or RT (alone or combined with CT), and were diagnosed with stage IA grade 2/3 with lymph-vascular space invasion (LVSI), and stage IB with two or more uterine risks, including old age, histological grade 2 or 3, LVSI and stage II, were included. According to the postoperative adjuvant therapy, all eligible patients were divided into two groups: CT group and RT (RT±CT) group. The primary objective was to investigate overall survival (OS) and disease-free survival (DFS) between the CT and RT groups. Grade 3 or worse adverse events were also presented in the present study. RESULTS: A total of 145 eligible patients were included. Among these patients, 97 patients underwent adjuvant CT and 48 patients underwent adjuvant RT (RT±CT). The median follow-up was 47.2 months, and the five-year OS rate was 92.7% in the CT group and 88.6 % in the RT group [hazard ratio (HR): 0.81, 95% confidence interval (CI): 0.22-2.99). The 5-year DFS rate for the two groups was 85.7% and 80.2%, respectively (HR: 0.82, 95% CI: 0.33-2.05). The cumulative incidence of local-regional disease recurrence at 60 months of follow-up was 6.2% in the CT group and 6.3% in the RT group (HR=1.11; 95%CI: 0.28-4.35). The cumulative incidence of distant recurrence at 60 months of follow-up was 5.2% in the CT group and 10.4% in the RT group (HR=0.65; 95%CI: 0.19-2.24). Both groups of patients were well-tolerant, and the only grade 3 or worse adverse events were neutropenia and thrombocytopenia. CONCLUSION: There was no difference in efficacy for adjuvant CT or adjuvant RT (RT±CT) in high-risk, early-stage endometrioid endometrial carcinoma. CT exhibited a trend of reducing the distant relapse, although there was no significant difference, when compared with adjuvant RT (RT±CT).
Subject(s)
Carcinoma, Endometrioid/drug therapy , Carcinoma, Endometrioid/radiotherapy , Endometrial Neoplasms/drug therapy , Endometrial Neoplasms/radiotherapy , Neoplasm Staging , Outcome Assessment, Health Care , Adult , Aged , Carcinoma, Endometrioid/mortality , Carcinoma, Endometrioid/surgery , Chemoradiotherapy , Chemotherapy, Adjuvant , Endometrial Neoplasms/mortality , Endometrial Neoplasms/surgery , Female , Follow-Up Studies , Humans , Middle Aged , Recurrence , Retrospective Studies , RiskABSTRACT
Polydnavirus (PDV) is a parasitic factor of endoparasitic wasps and contributes greatly to overcoming the immune response of parasitized hosts. Protein tyrosine phosphatases (PTPs) regulate a wide variety of biological processes at the post-transcriptional level in mammals, but knowledge of PDV PTP action during a parasitoid−host interaction is limited. In this study, we characterized a PTP gene, CvBV_12-6, derived from Cotesia vestalis bracovirus (CvBV), and explored its possible regulatory role in the immune response of the host Plutella xylostella. Our results from qPCR show that CvBV_12-6 was highly expressed in hemocytes at an early stage of parasitization. To explore CvBV_12-6 function, we specifically expressed CvBV_12-6 in Drosophila melanogaster hemocytes. The results show that Hml-Gal4 > CvBV_12-6 suppressed the phenoloxidase activity of hemolymph in D. melanogaster, but exerted no effect on the total count or the viability of the hemocytes. In addition, the Hml-Gal4 > CvBV_12-6 flies exhibited decreased antibacterial abilities against Staphylococcus aureus. Similarly, we found that CvBV_12-6 significantly suppressed the melanization of the host P. xylostella 24 h post parasitization and reduced the viability, but not the number, of hemocytes. In conclusion, CvBV_12-6 negatively regulated both cellular and humoral immunity in P. xylostella, and the related molecular mechanism may be universal to insects.
Subject(s)
Moths , Polydnaviridae , Animals , Amino Acid Sequence , Drosophila melanogaster/virology , Monophenol Monooxygenase/metabolism , Moths/virology , Polydnaviridae/genetics , Polydnaviridae/metabolism , Protein Tyrosine Phosphatases/genetics , Protein Tyrosine Phosphatases/metabolism , Host-Pathogen InteractionsABSTRACT
OBJECTIVE: To elucidate the effects of the deleted in colorectal carcinoma (DCC) gene on proliferation of ovarian cancer cell line SKOV-3. METHOD: An exogenous recombinant eukaryotic expression vector pcDNA3.1(+)-DCC, containing human DCC cDNA coding sequences, was constructed and transfected into SKOV-3 cells (SKOV-3/DCC). The pcDNA3.1 (+) transfected cells (SKOV-3/Neo) and SKOV-3 cells were used as the positive and negative controls, respectively. Expressions of DCC mRNA and protein were analyzed by RT-PCR and immunocytochemical analysis, respectively. Cell growth was detected by soft agar colony formation assay and MTT assay. Flow cytometry and transmission electron microscopy were used to assess the effects of DCC on cell cycle distribution and ultrastructure, respectively. BALB/c mice were used to evaluate the effects of DCC on tumorigenicity in vivo. RESULTS: RT-PCR and immunocytochemical analysis revealed the exogenous DCC gene was successfully transfected into SKOV-3 cell lines and obtained permanent expression. The half maximal inhibitory concentration (IC50) of SKOV-3/DCC cells was significantly lower than that of SKOV-3 or SKOV-3/Neo cells (all P<0.05). DCC expression caused SKOV-3 cells to be arrested in G1 phase (78.0%), and electron microscopic analysis showed SKOV-3/DCC cells displayed typical morphological changes of apoptosis. Two mice xenografted with SKOV-3/DCC cells showed no tumor tumorigenecity. The tumor volume of BALB/c mice bearing SKOV-3/DCC cells (3.403 mm(3)) was smaller than that of SKOV-3 cells (9.206 mm(3)). CONCLUSION: DCC gene may play an important role in suppressing the growth of SKOV-3 cell line and inducing apoptosis.
Subject(s)
Ovarian Neoplasms/therapy , Receptors, Cell Surface/metabolism , Tumor Suppressor Proteins/metabolism , Animals , Apoptosis/genetics , Apoptosis/physiology , Cell Cycle/genetics , Cell Cycle/physiology , Cell Line, Tumor , Cell Proliferation , DCC Receptor , Female , Humans , Immunohistochemistry , Mice , Mice, Inbred BALB C , Mice, Nude , Microscopy, Electron, Transmission , Ovarian Neoplasms/genetics , Ovarian Neoplasms/metabolism , Receptors, Cell Surface/genetics , Transfection , Tumor Suppressor Proteins/genetics , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE: To study the efficacy and safety of estradiol and drospirenone tablets (Angeliq) in treatment of menopausal symptoms among postmenopausal Chinese healthy women. METHODS: Total 244 postmenopausal Chinese healthy women who had moderate to severe hot flushes were randomly assigned into estradiol and drospirenone (observation group, n = 183) or placebo group (n = 61) by the ratio of 3:1 for 16 weeks in this randomized multi-center double-blind placebo-controlled study. During the trial, the follow-up visits were conducted at week 4, 8, 12, 16 of treatment and 2 weeks after treatment respectively. Height, weight, vital signs, hot flushes, other relevant menopausal symptoms and vaginal bleeding were observed in each follow-up visit, while the clinical global impression scale was assessed at 16 weeks as well. RESULTS: It showed that hot flushes were reduced significantly more in observation group than that in placebo group (P < 0.01), although both treatments were effective. The absolute values of mean severity index of total hot flushes decreased by -0.6 ± 0.5 in observation group and -0.4 ± 0.4 in placebo group from baseline respectively, which reached significant difference (P < 0.05). However, the absolute values of mean severity index of moderate to severe hot flushes decreased by -0.6 ± 0.8 in observation group and -0.3 ± 0.6 in placebo group from baseline respectively, which had no significant difference (P > 0.05). After 16 weeks treatment, it also showed that estradiol and drospirenone had significant better efficacy than placebo on moderate to severe sweating, vaginal dryness and clinical global impression scale (P < 0.01). During the trial, blood pressure in observation group was stable. The rate of vaginal bleeding in observation group was higher than that in the placebo group, especially during the week 4 to week 8 when 48.9% (87/178) in observation group and 10.7% (6/56) in placebo group of patients bled. Although the cumulative amenorrhea rate of observation group was lower than that of placebo group in each cycle (28 days), it increased gradually along with duration of the treatment. The commonest adverse event in observation group was breast tenderness which accounted for 12.0% (22/183). The level of serum potassium was in the normal range in observation group mostly.Meanwhile, the other adverse events rate was low. Serious adverse events reported in this trial were assessed as not study drug related or as unlikely study drug related. CONCLUSION: Estradiol and drospirenone tablets which could effectively alleviate menopausal symptoms in postmenopausal Chinese healthy women is a novel hormone replacement therapy regimen with high safety and efficacy.
Subject(s)
Androstenes/therapeutic use , Estradiol/therapeutic use , Estrogen Replacement Therapy , Hot Flashes/drug therapy , Mineralocorticoid Receptor Antagonists/therapeutic use , Postmenopause , Aged , Androstenes/adverse effects , Androstenes/pharmacology , China , Double-Blind Method , Estradiol/adverse effects , Estradiol/pharmacology , Female , Humans , Middle Aged , Mineralocorticoid Receptor Antagonists/adverse effects , Mineralocorticoid Receptor Antagonists/pharmacology , Tablets , Treatment Outcome , Vaginal Diseases/drug therapyABSTRACT
Polydnaviruses (PDVs) are obligatory symbionts of parasitoid wasps and play an important role in suppressing host immune defenses. Although PDV genes that inhibit host melanization are known in Microplitis bracovirus, the functional homologs in Cotesia bracoviruses remain unknown. Here, we find that Cotesia vestalis bracovirus (CvBV) can inhibit hemolymph melanization of its host, Plutella xylostella larvae, during the early stages of parasitization, and that overexpression of highly expressed CvBV genes reduced host phenoloxidase activity. Furthermore, CvBV-7-1 in particular reduced host phenoloxidase activity within 12 h, and the injection of anti-CvBV-7-1 antibody increased the melanization of parasitized host larvae. Further analyses showed that CvBV-7-1 and three homologs from other Cotesia bracoviruses possessed a C-terminal leucine/isoleucine-rich region and had a similar function in inhibiting melanization. Therefore, a new family of bracovirus genes was proposed and named as C-terminal Leucine/isoleucine-rich Protein (CLP). Ectopic expression of CvBV-7-1 in Drosophila hemocytes increased susceptibility to bacterial repression of melanization and reduced the melanotic encapsulation of parasitized D. melanogaster by the parasitoid Leptopilina boulardi. The formation rate of wasp pupae and the eclosion rate of C. vestalis were affected when the function of CvBV-7-1 was blocked. Our findings suggest that CLP genes from Cotesia bracoviruses encoded proteins that contain a C-terminal leucine/isoleucine-rich region and function as melanization inhibitors during the early stage of parasitization, which is important for successful parasitization.
Subject(s)
Genes, Viral , Melanins , Moths , Pigmentation , Polydnaviridae , Animals , Drosophila melanogaster/parasitology , Drosophila melanogaster/virology , Hemolymph , Host-Parasite Interactions , Isoleucine , Larva , Leucine , Monophenol Monooxygenase , Moths/parasitology , Moths/virology , Polydnaviridae/genetics , Wasps/virologyABSTRACT
Odorant-binding proteins (OBPs) are crucial in insect's olfactory perception, which participate in the initial step of odorant molecules transporting from the external environment to olfactory receptor neurons. To better understand the roles for OBPs in olfactory perception in Cotesia vestalis, a solitary larval endoparasitoid of diamondback moth, Plutella xylostella, we have comprehensively screened the genome of C. vestalis, and obtained 20 CvesOBPs, including 18 classic OBPs and two minus-C OBPs. Motif-pattern analysis indicates that the motifs of C. vestalis OBPs are highly conserved in Hymenoptera. The results of tissue expression analysis show that five OBPs (CvesOBP1/11/12/14/16) are highly expressed in male antennae, whereas six other OBP genes (CvesOBP7/8/13/17/18/19) are significantly transcriptionally enriched in female antennae. The results of RNA interference experiments for three most highly expressed OBP genes (CvesOBP17/18/19) in female antennae demonstrate that they are likely involved in parasitic processes of female wasps, as the wasps take a longer time to target the hosts when they are knocked down.
Subject(s)
Insect Proteins , Receptors, Odorant , Wasps , Animals , Female , Insect Proteins/genetics , Larva , Male , Moths/parasitology , Phylogeny , Receptors, Odorant/genetics , Wasps/geneticsABSTRACT
BACKGROUND: PMS2 expression loss was reported in a variety of human. However, its importance has not been fully understood in cervical carcinoma. The aim of this study was to determine the expression of PMS2 in cervical carcinoma and evaluate the significance of mismatch repair gene PMS2 regulated by glycogen synthase kinase 3beta (GSK-3beta) in chemosensitivity. METHODS: We examined PMS2 and phosphorylated GSK-3beta(s9) expression in cervical carcinoma tissues using immunohistochemical staining. Furthermore, we detected PMS2 expression in HeLa cells and evaluate the interaction with GSK-3beta after transfection with GSK-3beta by small interference RNA (siRNA), co-immunoprecipitation and immunoblotting. We also evaluated the effect of PMS2 transfection on HeLa cells' chemosensitivity to cisplatin treatment. RESULTS: We found significant downregulation of PMS2 in cervical carcinoma, which was negatively associated with phosphorylated GSK-3beta (s9). Furthermore, we demonstrated GSK-3beta transfection was able to interact with PMS2 and enhance PMS2 production in HeLa cells, and increased PMS2 production was responsible for enhanced chemosensitivity. CONCLUSIONS: Our results provide the evidence that stabilization of PMS2 production by GSK-3beta was important to improve chemosensitization, indicating the significance of GSK-3beta-related PMS2 downregulation in the development of cervical carcinoma and in developing a potential strategy for chemotherapy.
Subject(s)
Adenosine Triphosphatases/metabolism , DNA Repair Enzymes/metabolism , DNA-Binding Proteins/metabolism , Gene Expression Regulation, Neoplastic , Glycogen Synthase Kinase 3/metabolism , Uterine Cervical Neoplasms/genetics , Uterine Cervical Neoplasms/therapy , Base Pair Mismatch , Cisplatin/pharmacology , DNA Repair , Female , Glycogen Synthase Kinase 3 beta , HeLa Cells , Humans , Immunohistochemistry/methods , Mismatch Repair Endonuclease PMS2 , Phosphorylation , RNA Interference , RNA, Small Interfering/metabolism , TransfectionABSTRACT
OBJECTIVE: To construct a latent human soluble tumor necrosis factor receptor I (hsTNFRI) using the latency associated protein (LAP) of transforming growth factor-beta1 (TGF-beta1) fused via a matrix metalloproteinase (MMP) cleavage site to hsTNFRI so as to detect the latent biological activity of LAP-MMP-hsTNFRI fusion protein. METHODS: A double-stranded deoxyoligonucleotide coding for MMP cleavage site was cloned into plasmid pcDNA3.1. LAP and hsTNFRI cDNA were then inserted into both two sides of MMP cleavage site. After being transferred by LAP-MMP-hsTNFRI fusion gene with liposome, the expression of fusion protein in COS-7 cells was detected by RT-PCR and Western blot. The inhibitory effect of fusion protein upon cytotoxicity of TNF-alpha was detected by methyl thiazolyl tetrazolium (MTT) assay before and after the fusion protein incubated in MMP or peritoneal fluid from endometriosis patients. RESULTS: The recombinant plasmid LAP-MMP-hsTNFRI-pcDNA3.1 was constructed successfully and was expressed effectively in COS-7 cells. The MTT assay showed that there was no difference in the mortality rate of L929 cells between LAP-MMP-hsTNFRI-pcDNA3.1 and empty vector transfection groups (P > 0.05). The mortality rates of L929 cells with 800 ng/L TNF-alpha in LAP-MMP-hsTNFRI-pcDNA3.1 transfection group after incubation with MMP or peritoneal fluid from endometriosis patients were (44.5 +/- 2.4)% and (33.8 +/- 1.9)% respectively. And it was lower than the pre-incubation period (58.1 +/- 2.4)% (P < 0.05). CONCLUSION: The biological activity of LAP-MMP-hsTNFRI fusion protein can be made latent by LAP and activated by peritoneal fluid from endometriosis. Thus a new method has been provided for a targeted therapy of endometriosis.
Subject(s)
Endometriosis/therapy , Receptors, Tumor Necrosis Factor, Type I/genetics , Receptors, Tumor Necrosis Factor, Type I/therapeutic use , Ascitic Fluid/cytology , Cell Line , Female , Genetic Vectors , Humans , Plasmids , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/therapeutic use , TransfectionABSTRACT
OBJECTIVE: to evaluate clinical efficacy and safety of levornidazole in the treatment of pelvic anaerobic infections. METHODS: a multicenter randomized controlled clinical study was conducted to evaluate clinical efficacy and safety of levornidazole. One hundred and fourty-three patients with pelvic anaerobic bacteria infection were classified into 70 cases treated by levornidazole in study group and 73 cases treated by Ornidazole in control group. Those patients in two groups were both administered at a dose of 0.5 g twice daily for 5 - 7 days. The rate of clinical efficacy, bacteria clearance and adverse effect were recorded and compared between two groups. RESULTS: at the endpoint, the rate of clinical efficacy were 80% (56/70) in study group and 81% (59/73) in control group, which did not reach significant difference (P > 0.05). The rate of bacteria clearance were 97% (36/37) in study group and 92% (22/24) in control group, which also did not reach significant difference (P > 0.05). The rate of adverse reaction of 3% (20/70) in study group was significantly lower than 22% (16/73) in control group (P < 0.05). CONCLUSION: it is effective and safe to treat pelvic anaerobic infections with levornidazole and sodium chloride injection.
Subject(s)
Anti-Infective Agents/therapeutic use , Bacteria, Anaerobic/drug effects , Bacterial Infections/drug therapy , Ornidazole/therapeutic use , Pelvic Infection/drug therapy , Adolescent , Adult , Anti-Infective Agents/administration & dosage , Anti-Infective Agents/adverse effects , Bacterial Infections/microbiology , Double-Blind Method , Female , Humans , Injections, Intravenous , Middle Aged , Ornidazole/administration & dosage , Ornidazole/adverse effects , Pelvic Infection/microbiology , Sodium Chloride/administration & dosage , Sodium Chloride/adverse effects , Sodium Chloride/therapeutic use , Treatment Outcome , Young AdultABSTRACT
OBJECTIVE: To investigate the expression of microRNA 27a (miR-27a) and relationship with drug resistance in human ovarian cancer A2780/Taxol cells. METHODS: A stem-loop-mediated real-time PCR was used to detect miR-27a expression in A2780 and A2780/Taxol cells. The cells were transfected with the mimics or inhibitors of miR-27a or negative control RNA (NC) by lipofectamine 2000. The expressions of MDR1 gene, P-glycoprotein (P-gp) and homeodomain-interacting protein kinase 2 (HIPK2) protein levels were measured by real-time PCR and western blot respectively. Methyl thiazolyl tetrazolium (MTT) assay was used to analyze drug sensitivity. Apoptosis analysis was measured by fluorescence activated cell sorter (FACS). RESULTS: (1) miR-27a was an average of 2.2-fold higher expression level in A2780/Taxol cells than that in A2780 cells, with a significant difference between the two groups (P<0.05). (2) A2780/Taxol cells transfection with inhibitors of miR-27a showed that the levels of MDR1 mRNA was decreased by 39%, P-gp protein level [(26+/-5)%] decreased than that in the NC group [(43+/-7)%], HIPK2 protein level [(30+/-6)%] increased than that in the NC group [(19+/-4)%], the 50% inhibition concentration (0.5 micromol/L) was less than that in the NC group (6.8 micromol/L), apoptosis rate [(32.5+/-3.6)%] was higher than that in the NC group [(5.6+/-2.1)%], and there were significant differences between two groups (all P<0.05). (3) Transfection of A2780 cells with mimics of miR-27a led to increase MDR1 mRNA expression by 121% as compared with one transfection with NC (P<0.05). CONCLUSION: The expression of miR-27a is upregulated in A2780/Taxol cells, which may regulate MDR1 and P-gp expression by targeting HIPK2.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , Antineoplastic Agents/pharmacology , Drug Resistance, Neoplasm , MicroRNAs/genetics , Ovarian Neoplasms/metabolism , Paclitaxel/pharmacology , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Apoptosis/drug effects , Carrier Proteins/genetics , Carrier Proteins/metabolism , Cell Line, Tumor , Female , Flow Cytometry , Gene Expression Regulation, Neoplastic , Humans , MicroRNAs/antagonists & inhibitors , MicroRNAs/metabolism , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/genetics , Ovarian Neoplasms/pathology , Polymerase Chain Reaction/methods , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , TransfectionABSTRACT
Polydnaviruses (PDVs), classified into two genera, bracoviruses (BVs) and ichnoviruses (IVs), are large, double-stranded DNA viruses, which are beneficial symbionts of parasitoid wasps. PDVs do not replicate in their infected lepidopteran hosts. BV circles have been demonstrated to be integrated into host genomic DNA after natural parasitization. However, the integrations of IV circles in vivo remain largely unknown. Here, we analyzed the integration of Diadegma semiclausum ichnovirus (DsIV) in the genomic DNA of parasitized Plutella xylostella hemocytes. We found that DsIV circles are present in host hemocytes with non-integrated and integrated forms. Moreover, DsIV integrates its DNA circles into the host genome by two distinct strategies, conservatively, and randomly. We also found that four conserved-broken circles share similar motifs containing two reverse complementary repeats at their breaking sites, which were host integration motifs (HIMs). We also predicted HIMs of eight circles from other ichnoviruses, indicating that a HIM-mediated specific mechanism was conserved in IV integrations. Investigation of DsIV circle insertion sites of the host genome revealed the enrichment of microhomologies between the host genome and the DsIV circles at integration breakpoints. These findings will deepen our understanding of the infections of PDVs, especially IVs.
ABSTRACT
OBJECTIVE: To assess roles of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) in the mechanisms of lymphangiogenesis in epithelial ovarian carcinoma. METHODS: (1) Expression of Prox1, a newly described lymphatic endothelial cell nucleus marker, VEGF-A, VEGF-C, VEGF-D and PDGF-A, PDGF-B, PDGF-C, PDGF-D were detected by RT-PCR in SKOV3 cell line and in 90 ovarian tissue samples, included 15 benign tumors, 10 borderline tumors, 45 malignant tumors and 20 normal ovarian samples. (2) Expression levels of Prox1, VEGF-A, -C, -D and PDGF-A, -B, -C, -D were detected in 90 ovarian tissue sample mentioned above by real-time quantitative PCR (RTQ-PCR). RESULTS: (1) Prox1 was expressed in ovarian samples mentioned above, while not detected in SKOV3 cell. VEGF-A, -C, -D and PDGF-A, -B, -C, -D were found in SKOV3 cell and various ovarian tissues. (2) Expression levels of Prox1 (2.2 +/- 1.3, P < 0.01), VEGF-A (3.5 +/- 1.5, P < 0.01), VEGF-C (19 +/- 14, P < 0.01), VEGF-D (3.0 +/- 1.8, P < 0.01) and PDGF-A (3.3 +/- 3.3, P < 0.05), PDGF-C (6.9 +/- 4.6, P < 0.01) in malignant group were found to be significantly higher than those in borderline group and benign group. (3) The expression levels of Prox1, VEGF-A and PDGF-A were significantly greater in samples from the patients with lymph node metastasis (Prox1: 3.0 +/- 1.4, VEGF-A: 4.1 +/- 1.7, PDGF-A: 4.9 +/- 4.1), peritoneum metastasis (Prox1: 2.8 +/- 0.9, VEGF-A: 4.0 +/- 1.8, PDGF-A: 4.5 +/- 4.0) and in stage III - IV (Prox1: 2.6 +/- 1.3, VEGF-A: 4.0 +/- 1.4, PDGF-A: 4.1 +/- 3.7) than those without lymph node metastasis, without peritoneum metastasis and in stage I - II. There was a significant increased in the degree of VEGF-C and VEGF-D expression in positive lymph node metastasis group (VEGF-C: 24 +/- 13, VEGF-D: 3.9 +/- 2.0) compared with negative group (P < 0.05). (4) There were significant positive correlations between the expression levels of Prox1 and VEGF-D (r = 0.62, P < 0.01), PDGF-C (r = 0.91, P < 0.01) or PDGF-D (r = 0.61, P < 0.01). CONCLUSIONS: VEGF-A, VEGF-C and PDGF-A may promote lymphatic metastasis in epithelial ovarian carcinoma through else mechanisms other than lymphangiogenesis. VEGF-D may facilitate lymphangiogenesis and lymph node metastasis in epithelial ovarian cancer. There is no significant correlation between the expression of PDGF-B and lymphangiogenesis and lymph node metastasis. PCGF-C and PDGF-D may motivate lymphangiogenesis, but could not participate in lymph node metastasis in ovarian carcinoma.