ABSTRACT
Blackleg of oilseed rape caused by Leptosphaeria maculans/L. biglobosa is a worldwide important disease. L. maculans is more virulent than L. biglobosa, so it causes a great concern for oilseed rape production. In China, blackleg (L. biglobosa) of oilseed rape was reported in the 2000s, but epidemiological features of blackleg have not been well elucidated. Moreover, whether L. maculans exists in China is still an open question. Therefore, a 5-year survey was done in China to collect blackleg-occurrence data for characterizing the features of blackleg epidemics and to identify the blackleg pathogens for assessing the risk of L. maculans invasion. The results showed that all the 19 surveyed provinces had blackleg on oilseed rape, and the most frequently occurring provinces are Gansu, Qinghai, Shaanxi, and Hubei. Phoma stem canker was the most common symptom, which was associated with stem cracks on winter oilseed rape and with stem-weevil activities on spring oilseed rape. Temperature and rainfall were the main factors for blackleg epidemics on winter oilseed rape, whereas rainfall was the main factor for blackleg epidemics on spring oilseed rape. Brassica campestris and B. juncea oilseed rapes were more susceptible than B. napus to blackleg. Oilseed rapes cultivated under the continuous dry land-cropping pattern were more prone to blackleg than those cultivated under the paddy land/dry land-cropping pattern. All 6,015 fungal isolates from blackleg plant tissues belonged to L. biglobosa. These results are helpful for understanding the blackleg epidemics of oilseed rapes and for management of this disease in China.
Subject(s)
Ascomycota , Brassica napus , Plant Diseases/microbiology , Brassica napus/microbiology , ChinaABSTRACT
Objective: To investigate the efficacy and safety of transcatheter aortic valve replacement (TAVR) in the treatment of severe aortic stenosis. Methods: The clinical data of patients with severe aortic stenosis who underwent TAVR at the People's Hospital of Xinjiang Uygur Autonomous Region between September 2016 and September 2022 were retrospectively analyzed. Changes in aortic transvalvular pressure gradients, valve orifice area, and activity tolerance of patients before and after the surgery were compared. Moreover, postoperative complications and follow-up results from 30 days to 6 years after the surgery were recorded. Results: A total of 76 patients were included in the study (50 males and 26 females), with an average age of (71.3±7.6) years, including 16 rheumatic valvular diseases, 60 senile degenerative diseases, 46 bicuspid valves and 30 tricuspid valves. The success rate of the operation was 96.1% (73/76). Compared with that before the operation, the mean aortic transvalvular pressure gradients decreased [(8.5±2.8) mmHg vs (68.5±19.2) mmHg (1 mmHg=0.133 kPa),P<0.001], but the valve orifice area increased [(1.91±0.31) cm2 vs (0.65±0.21) cm2, P<0.001]. Likewise, six-minute walking test (6MWT) showed that walking distance was longer after the surgery [(430±13) m vs (201±28) m, P<0.001]. There were 1 case of retroperitoneal hematoma, 1 case of stricture balloon dilatation after femoral artery suture concomitant with postoperative puncture site infection, 1 case of femoral artery surgical incision, 2 cases of valve-in-valve (ViV) and 5 cases of perivalvular leakage (4 cases were mild and 1 case was moderate) after the surgery, respectively. Moreover, acute left main artery occlusion during operation occurred in 1 case, ventricular rupture during operation occurred in 1 case and the patient was transferred to valve replacement surgery and finally dead, delayed coronary artery occlusion and death happened in 1 case, and all of the above-mentioned 3 cases were due to surgical failure. Postoperative pacemaker implantation due to third-degree atrioventricular block was performed in 5 cases. There were 1 case of pulmonary embolism, 1 case of transient right limb disorder, 1 case of gastrointestinal bleeding and 1 case of urethral bleeding after the surgery, respectively. The patients were followed up for (1.0±0.1) years (30 days to 6 years), and the results showed that 2 cases died, including 1 case died suddenly at home (the cause of death was unknown) and 1 case died of acute heart failure 8 months after the surgery. Chronic heart failure occurred repeatedly in 6 cases. The quality of life of other patients improved significantly. Conclusion: TAVR is effective and safe for patients with severe aortic stenosis.
Subject(s)
Aortic Valve Stenosis , Heart Valve Prosthesis Implantation , Heart Valve Prosthesis , Transcatheter Aortic Valve Replacement , Male , Female , Humans , Middle Aged , Aged , Transcatheter Aortic Valve Replacement/methods , Aortic Valve/surgery , Aortic Valve Stenosis/surgery , Retrospective Studies , Quality of Life , Treatment Outcome , Postoperative Complications , Risk FactorsABSTRACT
Objective: To explore the clinical characteristics of various types of infected pancreatic necrosis(IPN) and the prognosis of different treatment methods in the imaging classification of IPN proposed. Methods: The clinical data of 126 patients with IPN admitted to the Department of Pancreatic and Biliary Surgery, the First Affiliated Hospital of Harbin Medical University from December 2018 to December 2021 were analyzed retrospectively. There were 70 males(55.6%) and 56 females(44.4%), with age(M(IQR)) of 44(17)years (range: 12 to 87 years). There were 67 cases(53.2%) of severe acute pancreatitis and 59 cases (46.8%) of moderately severe acute pancreatitis. All cases were based on the diagnostic criteria of IPN. All cases were divided into Type â (central IPN)(n=21), Type â ¡(peripheral IPN)(n=23), Type â ¢(mixed IPN)(n=74) and Type â £(isolated IPN)(n=8) according to the different sites of infection and necrosis on CT.According to different treatment strategies,they were divided into Step-up group(n=109) and Step-jump group(n=17). The clinical indicators and prognosis of each group were observed and analyzed by ANOVA,t-test,χ2 test or Fisher exact test,respectively. Results: There was no significant difference in mortality, complication rate and complication grade in each type of IPN(all P>0.05). Compared with other types of patients, the length of stay (69(40)days vs. 19(19)days) and hospitalization expenses(323 000(419 000)yuan vs. 60 000(78 000)yuan) were significantly increased in Type â £ IPN(Z=-4.041, -3.972; both P<0.01). The incidence of postoperative residual infection of Type â £ IPN was significantly higher than that of other types (χ2=16.350,P<0.01). There was no significant difference in the mortality of patients with different types of IPN between different treatment groups. The length of stay and hospitalization expenses of patients in the Step-up group were significantly less than those in the Step-jump group(19(20)days vs. 33(35)days, Z=-2.052, P=0.040;59 000(80 000)yuan vs. 122 000(109 000)yuan,Z=-2.317,P=0.020). Among the patients in Type â £ IPN, the hospitalization expenses of Step-up group was significantly higher than that of Step-jump group(330 000(578 000)yuan vs. 141 000 yuan,Z=-2.000,P=0.046). The incidence of postoperative residual infection of Step-up group(17.4%(19/109)) was significantly lower than that of Step-jump group(10/17)(χ2=11.980, P=0.001). Conclusions: Type â £ IPN is more serious than the other three types. It causes longer length of stay and more hospitalization expenses. The step-up approach is safe and effective in the treatment of IPN. However, for infected lesions which are deep in place,difficult to reach by conventional drainage methods, or mainly exhibit "dry necrosis", choosing the step-jump approach is a more positive choice.
Subject(s)
Intraabdominal Infections , Pancreatitis, Acute Necrotizing , Male , Female , Humans , Retrospective Studies , Pancreatitis, Acute Necrotizing/diagnosis , Pancreatitis, Acute Necrotizing/therapy , Pancreatitis, Acute Necrotizing/complications , Acute Disease , Intraabdominal Infections/complications , Necrosis/complications , Treatment OutcomeABSTRACT
Objective: To investigate the role and mechanism of cell adhesion molecule L1 like (CHL1) in insulin resistant adipocytes and insulin resistant mouse model induced by high glucose and high fat. Methods: The 3T3-L1 preadipocytes were randomly divided into control group (transfected with empty vector) and CHL1 overexpression group (transfected with CHL1 vector), cells were then induced to mature adipocytes by insulin, and insulin resistance was then induced by high sugar and high fat. The glucose content was measured to determine the glucose consumption of cells from the two groups. Protein expression levels of CHL1 and glucose transporter 4 (GLUT4), serine/threonine protein kinase (AKT) phosphorylation levels were detected by Western blot (WB), the mRNA expression levels of TNF-α and IL-6 were detected by real-time quantitative PCR (RT-qPCR). 24 C57BL/6 adult male mouse were randomly divided into conventional diet group (regular group), high-fat diet group (high-fat group), empty vector overexpression+high-fat group and CHL1 overexpression+high-fat group (n=6 each group). CHL1 overexpression was induced by tail vein injection of lentivirus. Four months later, mice were sacrificed, body weight was determined, and the epididymal white adipose tissue was collect. Hematoxylin-eosin staining (HE) was used to observe the pathology of mouse epididymal white adipose tissue, the expression of CHL1 was evaluated by immunohistochemical staining(IHC), RT-qPCR was used to detect the mRNA expression levels of CHL1, TNF-α and IL-6 in mouse epididymal white adipose tissue. Results: In vitro, glucose consumption was significantly higher in the CHL1 overexpression group than in the control group (P<0.05), and the protein expressions of CHL1 and GLUT4 were higher in the CHL1 overexpression group than those in the control group (P<0.01), and the mRNA expressions levels of TNF-α and IL-6 were lower in the CHL1 overexpression group than those in the control group (P<0.01). In vivo, the body weight and epididymal white adipose tissue of mouse were higher in the high-fat group and the empty vector overexpression+high-fat group than those in the conventional group (P<0.01), which were lower in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.01). HE results showed that the volume of epididymal white adipocytes was larger in the high-fat group and the overexpression control+high-fat group than that in the conventional group, which was smaller in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.01). The mRNA expression levels of IL-6 and TNF-α in epididymal white adipose tissue of mice were higher in the high-fat group and the empty vector overexpression+high-fat group than those in the conventional group (P<0.01), which were lower in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.05). IHC results showed that protein expression of CHL1 in epididymal white adipose tissue was lower in the high-fat group and the empty vector overexpression+high-fat group than in regular group, which was upregulated in the CHL1 overexpression+high fat group than in the empty vector overexpression+high-fat group (P<0.01). RT-qPCR results showed that mRNA expression of CHL1 in epididymal white adipose tissue was lower in the high-fat group and the empty vector overexpression+high-fat group than in regular group (P<0.01), which was higher in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.01). Conclusion: Overexpression of CHL1 can improve insulin resistance in adipocytes and mouse insulin resistance model induced by high glucose and high fat, and the beneficial effects might be mediated by the inhibition of AKT activation and the reduction of related inflammatory responses.
Subject(s)
Insulin Resistance , Insulin , Male , Mice , Animals , Tumor Necrosis Factor-alpha , Interleukin-6 , Proto-Oncogene Proteins c-akt , Mice, Inbred C57BL , Adipocytes , Disease Models, Animal , Glucose , Body Weight , RNA, Messenger , Cell Adhesion MoleculesABSTRACT
In recent years, with the withdrawal of paraquat (PQ) pesticides from the market, the number of poisoning cases caused by its substitute diquat (DQ) has shown an increasing trend year by year. Among the clinical manifestations of DQ poisoning, the damage to the central nervous system is relatively common and serious, but the specific toxicity mechanism is not clear, and there is no clear treatment. This article reviews the nervous system damage caused by DQ poisoning in order to improve the understanding systen of DQ poisoning.
Subject(s)
Herbicides , Poisoning , Diquat , Humans , Nervous System , ParaquatABSTRACT
Quetiapine is a psychotropic drug. Excessive use of quetiapine may lead to drowsiness, blurred vision, respiratory depression, hypotension and extrapyramidal reactions. Acute respiratory distress syndrome (ARDS) is rare due to overdose of quetiapine. On 14 February 2020, a patients with coma, respiratory arrest and hypotension due to overdose of quetiapine were admitted to our hospital. After receiving mechanical ventilationãplasma adsorption and anti-inflammatory treatment, the patient's consciousness turned clear, the machine was successfully removed and extubated, and the patient's condition was improved and discharged from hospital. We analyzed the clinical data of the patient with quetiapine poisoning, and discussed the clinical symptoms and chest CT characteristics of ARDS caused by quetiapine poisoning, in order to improve the understanding of quetiapine poisoning and improve the success rate of rescue.
Subject(s)
Antipsychotic Agents , Drug Overdose , Respiratory Distress Syndrome , Dibenzothiazepines , Drug Overdose/therapy , Humans , Quetiapine Fumarate/therapeutic useABSTRACT
In Drosophila melanogaster, ecdysis triggering hormone (ETH) is the key factor triggering ecdysis behaviour and promoting trachea clearance. However, whether ETH plays the dual roles in non-dipteran insects is unknown. In this survey, we found that Ldeth mRNA levels were positively correlated with circulating 20-hydroxyecdysone (20E) titers in Leptinotarsa decemlineata. Ingestion of an ecdysteroid agonist halofenozide or 20E stimulated the transcription of Ldeth, whereas RNA interference (RNAi) of ecdysteroidogenesis (LdPTTH or LdSHD) or 20E signalling (LdEcR, LdUSP or LdFTZ-F1) genes inhibited the expression, indicating ETH acts downstream of 20E. RNAi of Ldeth at the final instar stage impaired pupation. More than 80% of the Ldeth-depleted beetles remained as prepupae, completely wrapped in the old larval cuticles. These prepupae became withered, dried and darkened gradually, and finally died in soil. The remaining Ldeth hypomorphs pupated and emerged as abnormal adults, bearing smaller and wrinkle elytrum and hindwing. Moreover, the tracheae in the Ldeth hypomorphs were full of liquid. We accordingly proposed that the failure of trachea clearance disenabled air-swallowing after pupa-adult ecdysis and impacted wing expansion. Our results suggest that ETH plays the dual roles, initiation of ecdysis and motivation of trachea clearance, in a coleopteran.
Subject(s)
Benzoates/administration & dosage , Coleoptera/growth & development , Ecdysterone/administration & dosage , Hydrazines/administration & dosage , Molting/physiology , RNA Interference , Animals , Ecdysterone/antagonists & inhibitors , Insecticides/administration & dosage , Larva/growth & development , Pupa/growth & developmentABSTRACT
Objective: Based on the genetic diagnosis and follow-up study on pediatric neurofibromatosis 1 (NF1) patients, interrogating the genotype-phenotype correlations of patients with NF1 mutations. Methods: 32 Patients from age of 2 months to 5 years old (17 male and 15 female) suspected for neurofibromatosis 1 were recruited during September 2016 to January 2018 in Shanghai Children's Medical Center retrospectively. Genetic diagnosis was applied to detect pathogenic variants. Long-term follow-up study were conducted to reveal progress of the disease and genotype-phenotype correlations. Results: 27 patients were detected with pathogenic NF1 variants, among them three were not reported. 3 patients inherited pathogenic variants from their NF1 diagnosed parents, all the other variants were de novo. Progressive development of phenotypes wasn't observed in most patients during the follow-up (14/27). Some patients were diagnosed with short stature, pulmonary artery stenosis and developmental delay during the follow-up(7/27). Short stature and pulmonary artery stenosis may be associated with missense mutation and severe truncation mutation of NF1 gene, respectively. Conclusions: Genetic diagnosis is required in young patients of NF1.Follow-up plan of pediatric patients should be adjusted based on genetic findings. Early follow-up of cardiovascular abnormalities should be noted in patients with missense mutation. Height development in patients with severe truncating variants are needed.
Subject(s)
Neurofibromatosis 1 , Child , China , Female , Follow-Up Studies , Humans , Male , Neurofibromatosis 1/diagnosis , Neurofibromatosis 1/genetics , Neurofibromin 1 , Retrospective StudiesABSTRACT
Among the staff of Beijing Chao-Yang Hospital, Capital Medical University, who received the inactivated SARS-CoV-2 vaccine on January 30 in 2021, 28 recipients were selected for this research. Samples for nucleic acid tests were collected from the surface of the recipients' both hands before and after vaccination. The hemostatic stickers used after the inoculation were also collected for nucleic acid tests. The nucleic acid tests of the samples collected from the surface of both hands of the 28 recipients before vaccination were all negative. After vaccination, the nucleic acid tests of the samples collected from the surface of both hands of recipients were positive in 3 cases, and suspicious in 8 cases, with a positive rate of 10.7%. A total of 25 hemostatic stickers used were collected, 24 of them had positive nucleic acid tests, and the rest one had suspicious nucleic acid test result, with a positive rate of 96%. The hemostatic stickers used after the inoculation have the risk of nucleic acid contamination.
Subject(s)
COVID-19 , Hemostatics , Nucleic Acids , COVID-19 Vaccines , Humans , SARS-CoV-2 , VaccinationABSTRACT
Objective: To investigate the molecular mechanisms of clear cell renal cell carcinoma (CCRCC) with sarcomatoid differentiation (CCRCCS) and to explore new therapeutic targets for CCRCCS. Methods: Whole exome sequencing was performed on the carcinomatous and sarcomatoid components of five CCRCCS cases collected from January 2017 to October 2018. A highly frequent non-synonymous mutation of cadherin 23 (CDH23) was revealed by whole exome sequencing and further studied in additional samples. The sequencing of CDH23 in 40 specimens with CCRCCS and 50 specimens with CCRCC collected from January 2008 to October 2018 were conducted using Sanger sequencing. Immunohistochemistry was carried out to detect the protein expression of CDH23 in the additional 90 cases. Results: Carcinomatous and sarcomatoid components of CCRCCS shared most of the somatic single-nucleotide variants (SSNVs) as revealed through whole exome sequencing, while the sarcomatoid component had higher overall SSNVs than carcinomatous component. A highly frequent non-synonymous mutation of CDH23 (p.Arg1804Gln) was observed both in carcinomatous and sarcomatoid components of CCRCCS that resulted in the alteration in the highly conserved calcium-binding site mediating the functions of cadherins. In the additional 90 specimens, CDH23 mutation was much frequently detected in CCRCCS than that in CCRCC samples and even the high grade CCRCC. CDH23 protein was not or weakly expressed in most CCRCCS specimens with CDH23 mutation. There was an correlation between CDH23 gene mutation and negative expression of its protein (r=0.598, P<0.01). Conclusions: The present study reveals, for the first time, that the mutation of CDH23 (p.Arg1804Gln) is a genetic risk factor for CCRCCS. It is associated with the decreased expression of CDH23 protein, resulting in the absence of cadherin function of CDH23, indicating that CDH23 mutation may be involved in the sarcomatoid transformation in CCRCCS. Thus, CDH23 might be a potential therapeutic target for CCRCCS.
Subject(s)
Carcinoma, Renal Cell , Kidney Neoplasms , Cadherins/genetics , Carcinoma, Renal Cell/genetics , Cell Differentiation , Humans , Kidney Neoplasms/genetics , Exome SequencingABSTRACT
Objective: To analyze the pathogen distribution and drug resistance in acute,delayed and chronic periprosthetic joint infection (PJI). Methods: The clinical data of 316 patients with periprosthetic infection after primary hip and knee arthroplasty admitted to the Department of Arthroplasty,the First Affiliated Hospital,Xinjiang Medical University from August 2010 to August 2020 were retrospectively analyzed.There were 146 males and 170 females,aged (62.3±14.2) years (range:22 to 89 years).One hundred and sixty one patients underwent total hip arthroplasty and 155 patients underwent total knee arthroplasty.According to the time of postoperative infection,the patients were divided into acute PJI group (65 cases),delayed PJI group (83 cases) and chronic PJI group (168 cases).The results of pathogen species,composition ratio and drug susceptibility tests were collected,and the independent sample t test,Chi-square test or Fisher's exact probability test were used for comparison. Results: Gram-positive bacteria were the main pathogens of PJI (49.7%,157/316),and the positive rates of culture in patients with acute PJI,delayed PJI and chronic PJI were 33.8% (22/65),55.4% (46/83) and 53.0% (89/168),and the difference was statistically significant(χ²=8.343,P=0.015).The common bacteria were coagulase-negative Staphylococcus (54.8%,86/157) and Staphylococcus aureus (30.6%,48/157),The drug-sensitivity to linezolid,vancomycin and tigacycline was 100%.The gram-negative bacteria were mainly Escherichia coli and Enterobacter cloacae,and the drug resistance rate to carbapenems was low,ranging from 0 to 9.09%.The drug resistance rates of acute PJI patients to rifampicin,ciprofloxacin and erythromycin were significantly higher than those of late onset and chronic PJI patients,the difference was statistically significant(rifampicin:χ²=14.332,P=0.001;ciprofloxacin:χ²=12.086,P=0.002;erythromycin:χ²=9.096,P=0.010);The drug resistance rate of acute PJI patients to levofloxacin,clindamycin and tetracycline was higher than that of chronic PJI patients,and the difference was statistically significant(levofloxacin:χ²=10.500,P=0.002; clindamycin: χ²=7.103,P=0.007; tetracycline: χ²=6.909,P =0.012).The resistance rate of ampicillin/sulbactam in acute PJI (60.0%) was significantly higher than that in chronic PJI (16.7%),and the difference was statistically significant(χ²= 5.853,P=0.040). Conclusion: Gram-positive bacteria are the main pathogens of PJI,and the resistance rate of pathogens of acute PJI is higher than that of late onset and chronic PJI.
Subject(s)
Arthroplasty, Replacement, Hip , Prosthesis-Related Infections , Anti-Bacterial Agents/therapeutic use , Arthroplasty, Replacement, Hip/adverse effects , Drug Resistance, Bacterial , Female , Gram-Positive Bacteria , Humans , Male , Microbial Sensitivity Tests , Prosthesis-Related Infections/drug therapy , Retrospective StudiesABSTRACT
Objective: To further improve the awareness of the clinical feature of acute diquat poisoning. Methods: A retrospective analysis was performed on 4 cases of acute diquat poisoning with epileptoid seizure as the main clinical manifestation, which were admitted and diagnosed by the Characteristic Medical Center of Chinese People's Armed Police Force from January 1, 2017 to December 31, 2019. Take "Diquat" or "Deiquat" or "Reward" as keyword search for CNKI, Pubmed, and EMbase in both Chinese and English. The date of literature retrieval was from the database founding to December 31, 2019. Results: Of the 4 patients, 3 were male and 1 female, with an average age of 28 years (22-33 years) . The estimated dose was 8-20 g. All patients were treated with gastric lavage, catharsis, fluid replacement, blood perfusion, and in the early stage of treatment of epileptic seizures, the initial routine antiepileptic drugs had poor effect. Then propofol and midazolam were injected into the treatment. The epilepsy was relieved, but the condition deteriorated rapidly, and the patients died eventually. The literature search retrieved 3 patients in the 3 literatures included in the study were analyzed, and their clinical course was similar to that of 4 cases in the center. Necropsy was performedon all 3 patients, and the results were cerebral edema, diffuse cerebral edema, and hemorrhage around the basal ganglia. Conclusion: Acute diquat poisoning can cause epileptic seizures. Once it occurs, the disease progresses rapidly and the prognosis is poor. The combination of large dose of sedative drugs can be used to treat epilepsy in order to improve the prognosis.
Subject(s)
Epilepsy , Poisoning , Adult , Diquat , Female , Gastric Lavage , Humans , Male , Retrospective Studies , SeizuresABSTRACT
Henosepilachna vigintioctopunctata is a serious pest of Solanaceae and Cucurbitaceae in many Asian countries. RNA interference (RNAi) can effectively reduce transcript abundance in this beetle, offering opportunities to explore the biological function of specific genes. The white gene encodes a half-type ATP-binding cassette transporter that plays an essential role in tryptophan, guanine and uric acid transport across membranes. Mutations that disrupt the function of white are known to cause eye pigmentation phenotypes in many insect species. Here, we found evidence for five white gene paralogues present in H. vigintioctopunctata transcriptome datasets sequenced from a range of developmental stages. We individually knocked down each of the five white genes through the injection of corresponding double-stranded RNAs (dsRNAs) to the fourth-instar larvae to determine whether functional divergence has occurred. We found that injecting 1 µg dswhite3 caused compound eye colour of pupae and adults to develop as red/brown and brown, respectively, compared with black eyes in control beetles. Injection of 2 µg dswhite3 increased RNAi efficacy and produced a clearer eye colour phenotype. At both doses, the ocular diaphragm (a ring of black pigment surrounding each eye) did not change in the white3 RNAi hypomorphs. Moreover, our data revealed that injection of dswhite2 at the fourth-instar larval stage impaired the climbing ability of both male and female adults. Our results confirmed, for the first time, functional divergence of duplicated white genes in an insect species.
Subject(s)
Coleoptera/genetics , Insect Proteins/genetics , RNA Interference , Amino Acid Sequence , Animals , Coleoptera/growth & development , Coleoptera/metabolism , Female , Insect Proteins/chemistry , Insect Proteins/metabolism , Larva/genetics , Larva/growth & development , Larva/metabolism , Male , Phylogeny , Pigmentation/genetics , Pupa/genetics , Pupa/growth & development , Pupa/metabolism , RNA, Double-Stranded/administration & dosage , Sequence Alignment , TranscriptomeABSTRACT
Glutathione S-transferases (GSTs) are a detoxifying enzyme family that is essential for parasite blood-feeding and survival, and represent potential targets for hookworm vaccine development. Multiple GST-encoding complementary DNAs (cDNAs) have been cloned from Ancylostoma caninum and Necator americanus, but there are no reports about the cloning of this enzyme from Ancylostoma ceylanicum, the animal-derived zoonotic hookworm. To study the molecular nature and tissue localization of GST of A. ceylanicum (Ace-GST), we designed primers based on the GST gene sequence of A. ceylanicum in GenBank, amplified the Ace-GST cDNA by reverse transcription polymerase chain reaction, and analysed its homology and genetic evolution relationship. The amplified product was cloned into the pET-32a vector and transformed into Escherichia coli BL21 (DE3) for expression. To prepare anti-GST polyclonal antibodies, the recombinant protein was purified and used to immunize Kunming mice. The level of immunoglobulin G (IgG) antibody in the serum of immunized mice was detected by indirect enzyme-linked immunosorbent assay, and the Ace-GST localization in adult worm was determined using the immunofluorescence method. The results showed that the full-length cDNA encoding Ace-GST was 468 bp, which had the highest homology with Ac-GST-1 (60.1%) and clustered into one branch (v-class) with Ac-GST-1 and Na-GST-1 in a phylogenetic tree. Mice immunized with recombinant Ace-GST showed specific IgG antibody response. Immunolocalization revealed that natural Ace-GST is mainly located in the epidermis, muscle and intestine of the adult. These results may lay a foundation for further studies on the biological function of Ace-GST.
Subject(s)
Ancylostoma , Glutathione Transferase/metabolism , Ancylostoma/genetics , Ancylostoma/immunology , Ancylostoma/metabolism , Ancylostomiasis , Animals , Antibodies, Helminth , Antigens, Helminth/genetics , Antigens, Helminth/metabolism , Cloning, Molecular , DNA, Helminth/genetics , DNA, Helminth/isolation & purification , Escherichia coli/genetics , Glutathione Transferase/genetics , Immunohistochemistry , Phylogeny , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Transformation, BacterialABSTRACT
Objective: To study the clinicopathological features, immunophenotypes and MED12 gene status in benign metastasizing leiomyoma (BML). Methods: Nine cases of BML diagnosed at the Affiliated Hospital of Qingdao University from 2012 to 2018 were collected, and the radiologic and histologic features were analyzed. The protein expression of leiomyosarcoma-related driver genes, including RB1, PTEN,ATRX,p16,p53, as well as ER,PR,CD34,FH, and Ki-67 were detected using immunohistochemistry, and the mutation status of MED12 gene exon 2 was detected by Sanger sequencing. Results: All the nine patients with BML were female, and the age range was 48 to 64 years (median 55 years). All patients had history of uterine fibroids. The morphologic features of BML were similar to a benign uterine leiomyoma and did not exhibit malignant characteristics. All cases were positive for ER and PR, and negative for CD34. In addition, RB1, PTEN, ATRX, and FH were positive in all cases (wild type), while p16 showed a focally positive pattern. P53 positive index was less than 5% (wild type), and Ki-67 positive index was less than 1%. Sanger sequencing was done in six BML samples; one sample harbored a nonsense mutation c. 142_144delinsTAA (p.Glu48Ter), and another exhibited a synonymy mutation (c.192C>T, p.Phe64=)and one missense mutation c.196C>T (p.Pro66Ser). Conclusions: The present study suggests that BML is a unique leiomyoma entity that is pathologically and genetically different from leiomyosarcomas and conventional uterine leiomyomas. Evaluating the genetic phenotype of BML, especially the expression of leiomyosarcoma-related driver genes protein and MED12 gene status, may be helpful in understanding the pathogenesis of BML and in its differentiation from leiomyosarcoma.
Subject(s)
Leiomyoma , Uterine Neoplasms , Female , Humans , Middle Aged , Mutation , PhenotypeABSTRACT
The human genome is not a linear structure, but a three-dimensional structure through complex folding and assembly. Chromosome structure capture technology can detect the three-dimensional construction of chromatin. Hi-C sequencing data of various tumors indicate that the chromatin topology associated domains changed during tumor progression and is related to copy number variation. In addition, transformation of the genomic compartment is related to gene expression. However, current researches on three-dimensional structures of tumoral chromatin are still in the stage of exploration, and some conclusions are too superficial to be applied to the clinic immediately, which requires further study.
Subject(s)
Chromatin , DNA Copy Number Variations , Molecular Conformation , Neoplasms/physiopathology , Chromatin/physiology , DNA Copy Number Variations/physiology , Genomics , HumansABSTRACT
ABSTRACT: Objective To explore the characteristics of individuals with mental disorders suspected of road traffic offences and to find their differences from normal offenders, in order to regulate mental disorder patients' driving activities and prevent road traffic offences. Methods One hundred and twenty-three cases of forensic psychiatry testimony of individuals suspected of road traffic offences between 2014 and 2019 from the West China Forensic Center of Sichuan Province were collected. Fisher exact probability test was used to compare the differences between offenders with mental disorders and without mental disorders in terms of demographic characteristics, criminological characteristics, psychiatric characteristics and criminal responsibilities. Results There was no statistical significance in the differences of demographic characteristics, vehicles and kinds of alcohol between the two groups ï¼P>0.05ï¼. The main type of road traffic related crimes committed by offenders with mental disorders was risky driving and were mainly evaluated as partial criminal responsibility, whereas most offenders without mental disorders committed crime of causing traffic casualties and all were evaluated as full criminal responsibility. There was statistical significance in the differences of the types of crime and the criminal responsibility rating between the two groups ï¼P<0.05ï¼. Meanwhile, patients with mental disorders were characterized by long course of disease and irregular treatment, and individuals diagnosed as having mental disorders caused by psychoactive substances accounted for a large proportion. Conclusion There are differences in the characteristics of road traffic-related crimes between mental disorder patients and normal people. It is of great practical significance for reducing road traffic offences to evaluate whether the individuals with mental disorders are fit for driving.
Subject(s)
Automobile Driving , Criminals , Mental Disorders , China/epidemiology , Crime , Forensic Psychiatry , Humans , Mental Disorders/diagnosis , Mental Disorders/epidemiologyABSTRACT
Plantation-grown Eucalyptus (Myrtaceae) and other trees residing in the Myrtales have been widely planted in southern China. These fungal pathogens include species of Cryphonectriaceae that are well-known to cause stem and branch canker disease on Myrtales trees. During recent disease surveys in southern China, sporocarps with typical characteristics of Cryphonectriaceae were observed on the surfaces of cankers on the stems and branches of Myrtales trees. In this study, a total of 164 Cryphonectriaceae isolates were identified based on comparisons of DNA sequences of the partial conserved nuclear large subunit (LSU) ribosomal DNA, internal transcribed spacer (ITS) regions including the 5.8S gene of the ribosomal DNA operon, two regions of the ß-tubulin (tub2/tub1) gene, and the translation elongation factor 1-alpha (tef1) gene region, as well as their morphological characteristics. The results showed that eight species reside in four genera of Cryphonectriaceae occurring on the genera Eucalyptus, Melastoma (Melastomataceae), Psidium (Myrtaceae), Syzygium (Myrtaceae), and Terminalia (Combretaceae) in Myrtales. These fungal species include Chrysoporthe deuterocubensis, Celoporthe syzygii, Cel. eucalypti, Cel. guangdongensis, Cel. cerciana, a new genus and two new species, as well as one new species of Aurifilum. These new taxa are hereby described as Parvosmorbus gen. nov., Par. eucalypti sp. nov., Par. guangdongensis sp. nov., and Aurifilum terminali sp. nov. Pathogenicity tests showed that the eight species of Cryphonectriaceae are pathogenic to two Eucalyptus hybrid seedlings, Melastoma sanguineum branches, and Psidium guajava and Syzygium jambos seedlings. The overall data showed that Chr. deuterocubensis is the most aggressive, followed by Par. eucalypti. Significant differences in tolerance were observed between the two tested Eucalyptus hybrid genotypes, suggesting that disease-tolerant genotypes can be selected for disease management in the Eucalyptus industry.
ABSTRACT
Insect chitin deacetylases (CDAs) are carbohydrate esterases that catalyze N-deacetylation of chitin to generate chitosan, a process essential for chitin organization and compactness during the formation of extracellular chitinous structure. Here we identified two CDA2 splice variants (LdCDA2a and LdCDA2b) in Leptinotarsa decemlineata. Both splices were abundantly expressed in larval foregut, rectum, and epidermis; their levels peaked immediately before ecdysis within each instar. In vivo results revealed that the two isoforms transcriptionally responded, positively and negatively respectively, to 20-hydroxyecdysone and juvenile hormone signaling pathways. RNA interference (RNAi)-aided knockdown of the two LdCDA2 variants (hereafter LdCDA2) or LdCDA2b, rather than LdCDA2a, resulted in three negative effects. First, foliage consumption was significantly reduced, larval developing period was lengthened, and larval growth was retarded. Second, chitin contents were reduced, whereas glucose, trehalose, and glycogen contents were increased in the LdCDA2 and LdCDA2b RNAi larvae. Third, approximately 20% of LdCDA2 and LdCDA2b RNAi larvae were trapped within the exuviae and finally died. About 60% of the abnormal pupae died as pharate adults. Around 20% of the RNAi pupae emerged as deformed adults, with small size and wrinkled wings. These adults eventually died within 1 week after molting. Our results reveal that knockdown of CDA2 affects chitin accumulation. Consequently, LdCDA2 may be a potential target for control of L. decemlineata larvae.
Subject(s)
Amidohydrolases/metabolism , Coleoptera/enzymology , Molting , Amidohydrolases/genetics , Amino Acid Sequence , Animals , Chitin/metabolism , Coleoptera/genetics , Ecdysterone/metabolism , Gene Expression , Larva/enzymology , Pupa/physiology , RNA InterferenceABSTRACT
A heterodimer of ultraspiracle (USP) and ecdysone receptor (EcR) mediates 20-hydroxyecdysone (20E) signalling cascade to regulate insect moulting and metamorphosis. However, at least two questions remain to be addressed in terms of the molecular importance of USP in insect species. First, is USP involved in both regulation of ecdysteroidogenesis and mediation of 20E signalling in non-drosophilid insects, as in Drosophila melanogaster? Second, does USP play any role in larval metamorphosis except as the partner of heterodimeric receptor to activate the downstream 20E signalling genes? In this paper, we found that RNA interference (RNAi) of LdUSP in the final (fourth) instar larvae reduced the messenger RNA levels of four ecdysteroidogenesis genes (Ldspo, Ldphm, Lddib and Ldsad) and 20E titre, and repressed the expression of five 20E signal genes (EcRA, HR3, HR4, E74 and E75) in Leptinotarsa decemlineata. The LdUSP RNAi larvae remained as prepupae, with developing antennae, legs and discs of forewings and hindwings. Dietary supplement with 20E restored the expression of the five 20E signal genes, but only partially alleviated the decreased pupation rate in LdUSP RNAi beetles. Knockdown of LdUSP at the penultimate (third) instar larvae did not affect third-fourth instar moulting. However, silencing LdUSP caused similar but less severe impairments on pupation. Accordingly, we propose that USP is undoubtedly necessary for ecdysteroidogenesis, for mediation of 20E signalling and for initiation of metamorphosis in L. decemlineata.