Objective To investigate the prevalence of Schistosoma japonicum infections in wild rodents in schistosomiasis-endemic areas of China, so as to provide insights into formulation of technical guidelines for monitoring of and the precise control strategy for S. japonicum infections in wild rodents during the elimination phase. Methods Two administrative villages where schistosomiasis was historically highly prevalent were selected each from Dongzhi County, Anhui Province, and Duchang County, Jiangxi Province as study villages. Wild rodents were captured from study villages with baited traps or cages at night in June and September, 2021. The number of rodents captured was recorded, and the rodent species was characterized based on morphologi-cal characteristics. Liver tissues were sampled from captured rodents for macroscopical observation of the presence of egg granu- lomas, and S. japonicum infection was detected simultaneously using liver tissue homogenate microscopy, examinations of mesenteric tissues for parasites, and modified Kato-Katz thick smear technique (Kato-Katz technique). A positive S. japonicum infection was defined as detection of S. japonicum eggs or adult worms by any of these methods. The rate of wild rodent capture and prevalence of S. japonicum infections in wild rodents were compared in different study villages and at different time periods, and the detection of S. japonicum infections in wild rodents was compared by different assays. Results The overall rate of wild ro- dent capture was 8.28% (237/2 861) in Dongzhi County, and the wild rodent capture rates were 9.24% (133/1 439) and 7.31% (104/1 422) in two study villages (χ² = 3.503, P = 0.061), and were 8.59% (121/1 409) and 7.99% (116/1 452) in June and September, 2021, respectively (χ² = 0.337, P = 0.561). The overall rate of wild rodent capture was 3.72% (77/2 072) in Duchang County, and the wild rodent capture rates were 6.91% (67/970) and 0.91% (10/1 102) in two study villages (χ² = 51.901, P < 0.001), and were 4.13% (39/945) and 3.37% (38/1 127) in June and September, 2021, respectively (χ² = 0.815, P = 0.365). Rattus norvegicus was the predominant rodent species captured in both counties, accounting for 70.04% (166/237) of all captured wild rodents in Dongzhi County and 88.31% (68/77) in Duchang County. No S. japonicum infection was detected in wild rodents captured in Duchang County. Nevertheless, the overall prevalence of S. japonicum infections was 51.05% (121/237) in wild rodents captured in Dongzhi County, with prevalence rates of 50.38% (67/133) and 51.92% (54/104) in two study villages (χ² = 0.098, P = 0.755), and 54.31% (63/116) and 47.93% (58/121) in September and June, 2021, respectively (χ² = 0.964, P = 0.326). Of 237 wild rodents captured in Dongzhi County, there were 140 (59.07%) rodents with visible hepatic egg granulomas, 117 (49.47%) tested positive for S. japonicum eggs by liver tissue homogenate microscopy, 34 (14.35%) tested positive for S. japonicum eggs with Kato-Katz technique; however, no adult S. japonicum worms were detected in mesenteric tissues. In addition, hepatic egg granulomas were found in all wild rodents tested positive for S. japonicum eggs with liver tissue homogenate microscopy. Conclusions The rate of wild rodent capture and prevalence of S. japonicum infection in wild rodents vary greatly in schistosomiasis-endemic areas of China, and the prevalence of S. japonicum infection is slightly higher in wild rodents captured in autumn than in summer. Liver tissue is recommended as the preferred sample for surveillance of S. japonicum infection in wild rodents, and a combination of macroscopical observation of hepatic egg granulomas and liver tissue homogenate microscopy may be a standard method for surveillance of S. japonicum infection in wild rodents.

Objective:To establish a microwave digestion and inductively coupled plasma mass spectrometry(ICP-MS)for determination on calcium content of Xiao'er Magan granules and discuss the effect of decocting gypsum into medicine on calcium content.Methods:ICP-MS was adopted to determine the calcium content in the preparation,and the difference of calcium content in the small preparation and Xiao'er Magan granules prepared by different pul-verization degree gypsum and different alcohol precipitation processes were compared.Results:The consistency of calcium content between different enterprises and batches of the same enterprise was poor.The results showed that the larger the particles,the lower the calcium transfer rate.The smaller the particles,the higher the calcium trans-fer rate.Conclusion:The transfer rate of gypsum decocted calcium is related to its degree of comminution to ensure consistency of preparation quality.It is suggested to unify the comminution degree of gypsum.The method can pro-vide guidance for the optimization on preparation technology of Xiao'er Magan granules.

Objective To investigate the advantages and efficacy of traction with titanium clips in endoscopic submucosal dissection(ESD)for large laterally spreading tumor(LST)in rectum and sigmoid colon.Methods 67 patients with large sigmoid or rectal LST underwent ESD from January 2018 to June 2022 were analyzed retrospectively,including 32 patients in Group A and 35 patients in Group B.Group A was treated with clip-line traction and group B was treated with traditional ESD.The size of lesion,the total operation time,the submucosal dissection time,submucosal dissection rate,submucosal injection number,en bloc resection rate,R0 resection rate,curative resection rate and complications of the two groups were compared.Results LST-G-M was the most common type and villous adenoma was the main pathology in both groups.There were no differences in en bloc resection rate,R0 resection rate and incidence of complications between the two groups.The average size of group A was(13.6±8.4)cm2,significantly larger than that in group B(9.3±4.7)cm2,the total operation time was(42.3±10.3)min in group A,significantly shorter than that in group B(47.9±10.1)min,submucosal dissection time was(30.7±8.2)min in group A,significantly shorter than that in group B(36.1±7.6)min,submucosal injection number was(2.7±1.1)times in group A,significantly less than that in group B(3.5±1.2)times,submucosal dissection rate was(0.4±0.2)cm2/min in group A,significantly faster than that in group B(0.2±0.1)cm2/min,the differences were statistically significant(P<0.05).Conclusion Compared with traditional ESD,clip-line traction can provide a better surgical field and more effective dissection for large LST in rectum and sigmoid colon.

Objective To evaluate the effect of exercise-diet behavior intervention based on the transtheoretical model in patients undergoing weight loss surgery.Methods By convenience sampling,72 patients undergoing weight loss surgery in a tertiary general hospital in Xinxiang City,Henan Province from February 2021 to October 2022 were selected as the research subjects.By a random number table method,they were divided into a test group and a control group,with 36 cases in each group.The test group received exercise-diet behavior intervention based on the trans-theoretical model,while the control group received conventional intervention.The intervention began on the first day after admission and ended 6 months after surgery.The body mass index,body fat,lean body mass,diastolic blood pressure,systolic blood pressure,fasting blood glucose,insulin resistance index(HOMA-IR),Health Promoting Lifestyle Profile-Ⅱ(HPLP-Ⅱ)score,and Short-Form-36 Health Survey(SF-36)score were compared between the 2 groups before and after surgery for 3 and 6 months,as well as the complications within 6 weeks after surgery.Results The results of repeated measures analysis of variance showed that there was an interaction between the 2 groups in terms of anthropometric measurements,blood pressure and blood glucose,HPLP-Ⅱ scores,and SF-36 scores,with statistically significant differences(P＜0.001).After 6 months of surgery,the body mass index(23.32±2.32),body fat(24.10±3.46)kg,and lean body mass(41.64±3.24)kg in the test group were lower than(27.32±3.64),(28.46±4.18)kg,and(46.68±4.65)kg in the control group,and the differences were statistically significant(P＜0.001).At 3 and 6 months after operation,the diastolic blood pressure,systolic blood pressure,fasting blood glucose and HOMA-IR of the test group were lower than those of the control group,and the differences were statistically significant(P＜0.05).The HPLP-Ⅱ score of the test group was higher than that of the control group at 3 and 6 months after operation(P＜0.001).The SF-36 score of the test group was significantly higher than that of the control group at 6 months after operation(P＜0.05).The incidence of complications in the test group was 2.56％,which was not significantly different from 19.44％in the control group(P＞0.05).Conclusion The exercise-dietary behavior intervention based on the transtheoretical model can promote the formation of healthy behaviors in patients undergoing weight loss surgery,maintain weight loss effects,improve blood pressure and blood glucose levels,and enhance the quality of life of patients.

AIM To investigate the effects of Compound Dihuang Granules on biological function of dopaminergic neurons in rats with Parkinson's disease(PD)of Pattern of Yin-Deficiency with Stirring Wind.METHODS The PD animal model of Pattern of Yin-Deficiency with Stirring Wind established by intracerebral injection of 6-hydroxydopamine(6-OHDA)were randomly divided into the model group,the Midoba group(150 mg/kg)and the low,medium and high dose Compound Dihuang Granules groups(1.75,3.5 and 7 g/kg)for corresponding drug intervention,in contrast to the normal group,the sham operation group and the model group underwent 28-day normal saline administration.The rats had their general condition and neuroethology observed;their pathological changes of substantia nigra observed by HE staining;their mitochondrial structure of dopaminergic neurons in the damaged substantia nigra observed by transmission electron microscopy;and their expressions of DJ-1,IP3R,GRP75 and VDAC1 detected by immunohistochemistry,Western blot and RT-qPCR.RESULTS Compared with the normal group and sham operation group,the model group displayed increased rotational behavirors(P＜0.01),decreased swimming time score(P＜0.01),decreased hanging time(P＜0.01),decreased number of neurons in substantia nigra but more neurons with morphological damage,mitochondrial swelling and degeneration,mitochondrial crista disappearance,and decreased expressions of DJ-1,IP3R,GRP75 and VDA C1 mRNA and protein in the injured side(P＜0.01).Compared with the model group,the Midoba group and the Compound Dihuang Granules groups demonstrated less rotational behavirors(P＜0.01),higher swimming time score(P＜0.01),longer hanging time(P＜0.01),less damage to mitochondrial morphology and structure,and higher expressions of DJ-1,IP3R,GRP75 and VDA C1 mRNA and protein in the injured substantia nigra(P＜0.05,P＜0.01).The high-dose Compound Dihuang Granules presented equivalent efficacy to that of Midoba.CONCLUSION Compound Dihuang Granules may promote endoplasmic reticulum-mitochondria homeostasis,reduce mitochondrial damage and maintain the biological function of dopaminergic neurons by regulating the expressions of DJ-1,IP3R,GRP75 and VDAC1.

Objective To analyze the clinicopathological features, diagnosis and differential diagnosis of bronchiolar adenoma (BA) with primary lung cancer. Methods Data of 17 BA patients complicated with primary lung cancer in Dalian University Affiliated Xinhua Hospital from January 2020 to December 2022 were collected, and the clinical data, histopathological features and immunohistochemical features were summarized. Results 17 patients included 7 males and 10 females, with 49-82 years old. There were 2 squamous cell carcinoma and 15 adenocarcinoma. BA was mostly nodular, without capsule. It had a clear boundary with the surrounding lung tissue, with a major diameter of 0.3-1.5 cm. The section of BA was gray-white, gray-red, gray-brown, with medium-soft texture, and a few had obvious mucus. Microscopically, it was composed of luminal cell layer and basal cell layer, and luminal cells contain mucus cells, ciliated cells, cubic cells and columnar cells. According to the composition of luminal cells, BAs were divided into proximal type and distal type. There were 5 proximal type and 12 distal type, including 1 case of atypical distal BA. Immunohistochemistry results showed that in typical BAs, CK5/6, p63 and p40 were positive in basal cells, thyroid transcription factor-1 was positive in luminal cells and basal cells, and the proliferation index of Ki-67 was 1%. Conclusions BA is a benign tumor, and is a bilayer structure mainly composed of luminal cells and basal cells. However, when the absence or discontinuity of basal cells, BA is easily misdiagnosed as malignancy, which should be highly vigilant.

ObjectiveTo explore psychological and behavioral characteristics of children with autism spectrum disorder (ASD) using Psycho-educational Profile (Third Edition) (PEP3). MethodsFrom October, 2021 to October, 2022, 192 children with ASD without intervention in the Binzhou Medical University Hospital were selected as observation group, and 96 healthy children who visited at the same time were selected as control group. They were assessed with PEP3. ResultsThe development of receptive language significantly delayed behind expressive language (t = 5.383, P < 0.001) in the observation group. The cognitive, expressive language, receptive language, fine motor, gross motor, imitation and personal self-care correlated with age (r = 0.540 to 0.795, P < 0.001). The Cronbach's α of PEP3 for the observation group was 0.810 to 0.947. The original score of each subtest of PEP3 was less in the observation group than in the control group (|t| > 4.267, P < 0.001). ConclusionThe development of receptive language retards behind the expressive language in children with ASD. Cognitive and motor functions develop with age, while the correlation between maladaptive behavior and age is weak. PEP3 is reliable in internal consistency, and valid in discrimination.

Objective To investigate the clinical efficacy of drug eluting stents vs metal bare stents for femoral popliteal arteriosclerosis obliterans.Methods The clinical data of 47 patients with femoral popliteal arte-riosclerosis obliterans receiving endovascular therapy from October 2020 to June 2021 were retrospectively ana-lyzed.A total of 24 cases received drug-eluting stents(DES group)and 23 cases underwent metal bare stents(BMS group).Results All patients successfully completed the operation without any adverse events.There was no statistical difference in ABI between the DES group and the BMS group at 7 days and 3 months after surgery,and ABI growth value of the DES group was higher than that of the BMS group at 6,12 and 24 months after surgery(P<0.05).There was no significant difference in the primary patency rate at 6 months after surgery,However,the primary patency rate in DES group was higher than that in BMS group at 12 and 24 months after surgery(91.7%vs.65.2%,83.3%vs.56.5%,P<0.05).For the target lesion revascularization rate of the two groups,DES group had a significant advantage over BMS group(4.0%vs.26.1%,P<0.05).Conclusion DES had better clinical efficacy and advantage over bare metal stent for the treatment of femoral popliteal arteriosclerosis obliterans.

OBJECTIVE: To express and purify the antigenic peptide of adeno-associated virus (AAV) capsid conserved regions in prokaryotic cells and prepare its rabbit polyclonal antibody. METHODS: The DNA sequence encoding the conserved regions of AAV capsid protein was synthesized and cloned into the vector pET30a to obtain the plasmid pET30a-AAV-CR for prokaryotic expression and purification of the conserved peptides. Coomassie blue staining and Western blotting were used to identify the AAV conserved peptides. Japanese big ear white rabbits were immunized with AAV conserved region protein to prepare polyclonal antibody, with the rabbits injected with PBS as the control group. The antibody titer was determined with ELISA, and the performance of the antibody for recognizing capsid protein sequences of AAV1-AAV10 was assessed with Western blotting and immunofluorescence assay. RESULTS: The plasmid pET30a-AAV-CR was successfully constructed, and a recombinant protein with a relative molecular mass of 17000 was obtained. The purified protein induced the production of antibodies against the conserved regions of AAV capsid in rabbits, and the titer of the purified antibodies reached 1:320 000. The antibodies were capable of recognizing a wide range of capsid protein sequences of AAV1-AAV10. CONCLUSION We successfully obtained the polyclonal antibodies against AAV capsid conserved region protein from rabbits, which facilitate future studies of AAV vector development and the biological functions of AAV.
Animals ; Antibodies ; Capsid ; Capsid Proteins/genetics* ; Dependovirus/genetics* ; Prokaryotic Cells ; Rabbits ; Recombinant Proteins/genetics*

Aspergillus niger is an important industrial strain which has been widely used for production of enzymes and organic acids. Genome modification of A. niger is required to further improve its potential for industrial production. CRISPR/Cas9 is a widely used genome editing technique for A. niger, but its application in industrial strains modification is hampered by the need for integration of a selection marker into the genome or low gene editing efficiency. Here we report a highly efficient marker-free genome editing method for A. niger based on CRISPR/Cas9 technique. Firstly, we constructed a co-expression plasmid of sgRNA and Cas9 with a replication initiation region fragment AMA1 (autonomously maintained in Aspergillus) by using 5S rRNA promoter which improved sgRNA expression. Meanwhile, a strain deficient in non-homologous end-joining (NHEJ) was developed by knocking out the kusA gene. Finally, we took advantage of the instability of plasmid containing AMA1 fragment to cure the co-expression plasmid containing sgRNA and Cas9 through passaging on non-selective plate. With this method, the efficiency of gene editing reached 100% when using maker-free donor DNA with a short homologous arm of 20 bp. This method may facilitate investigation of gene functions and construction of cell factories for A. niger.
Gene Editing ; Aspergillus niger/genetics* ; CRISPR-Cas Systems/genetics* ; Plasmids/genetics*