Objective To provide the basis for the differentiation with similar species of intestinal flukes through observing the figure of Haplorchis pumilio. Methods Adults of H. pumilio were collected from the intestine of the cat which was infected with the encysted cercariae of H.pumilio for 45 days. The worms were observed after staining. Eggs and metacercariae of H.pumilio were collected and examined for their shape, size and morphological characteristics. Pseudorasbora parva, the fish host, was examined for the parasitized sites of metacercariae. Results The principal characteristics of the adults is the acetabulum degradation. There are only the genital sucker with 44-48 hamuli. The average measurement of eggs is 31.2?16.7 ?m with a smooth shell. Its aceromion is not evident. The average diameter of metacercariae is 168.5 ?m. There are squamous spines on metacercaria. The metacercariae only parasitize in the muscle between the basis of the fin and the fish body. The average measurement of metacercaria cyst is 445?95?m, with squamous spines on the body surface. Hamuli are found on the genital sucker of metacercaria cyst. Conclusion The morphological figures and parasitic sites of metacercaria, the genital sucker of the adult, and the number and form of the hamulus on the genital sucker provide basis for distinguishing H. pumilio from other intestinal flukes.

Objective To develop a method for detecting the genotype of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) alleles. Methods According to the sequence characteristic of PvMSP-1, nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to amplify the polymorphic region of ICB5-ICB6 which contains Q repeats and PvuII restriction site (Sal-1 type). The PCR product was digested by PvuII restriction endonuclease and the digested fragments were observed by 2% agarose gel electrophoresis. The allelic type was determined according to the banding pattern. Results Bands in size of 400 bp (Belem type ) and/or 470 bp (Sal-1 type ) appeared in all 98 P. vivax isolates, no band was found in negative control. After PvuII digestion, two Sal-1 type fragments (120 bp and 350 bp) were obtained from 45 samples of 470 bp. Single-band of 400 bp appeared in 3 of 40 samples with 400 bp as Belem type, two bands of 120 bp and 280 bp appeared from other 35 samples as recombination type III, and another 2 bands with 120 bp and 240 bp as Korean isolate. Conclusion The result showed that the nested PCR-RFLP may be applied in the detection and identification of the three PvMSP-1 allelic types in China.

Objective To develop methods of extracting DNA from malaria parasites on Giemsa-stained blood smears. Methods Improved Na2HPO4 method and Chelex-100 ion-exchange technique were used to extract DNA from Giemsa-stained or unstained blood smears. Nested PCR was employed for amplification and identification of allelotypes in the Plasmodium vivax merozoite surface protein-1(PvMSP-1). Results Target DNA bands appeared in all samples of unstained thick blood smears, while no DNA bands were visible in the fixed and stained thin smears. Both methods identified PvMSP-1 alleles from smears with parasitemia of ≥0.01%. Conclusion It is feasible to identify PvMSP-1 alleles from Giemsa-stained blood smear.

OBJECTIVE: To explore the genetic etiology and clinical outcome of a child with steroid-resistant nephrotic syndrome and diffuse mesangial sclerosis. METHODS: Genomic DNA was extracted from peripheral blood leukocytes of the proband and his parents. Targeted capture - next generation sequencing and Sanger sequencing were carried out. Candidate variant was verified by segregation analysis in his family. RESULTS: A heterozygous missense variant of the TRPC6 gene, namely c.325G>A (p.Gly109Ser), was detected in the proband. The same variant was not detected in either parent. According to the guidelines for the interpretation of sequence variants developed by American College of Medical Genetics and Genomics, the variant was predicted as pathogenic. CONCLUSION The missense variant of the TRPC6 gene probably underlay the diffuse mesangial sclerosis in this patient. Above finding has expanded the phenotypic spectrum of the TRPC6 gene.
Child ; Genomics ; Humans ; Mutation, Missense ; Nephrotic Syndrome/genetics* ; Sclerosis ; TRPC6 Cation Channel/genetics*

Objective　To detect the location of pathogens in myocardium using in situ RT-PCR technique in order to study the pathogenetic course of the myocardial lesion induced by CoxB3m virus infection in mice. Methods　(1) Thirty and fifty Balb/c mice were used respectively to establish the acute and chronic CoxB3m infected models, with another 25 healthy mice as the controls; (2) KS400 image analysis system (Germany) was used to measure the cardiac chamber area and the left ventricular wall thickness of the chronic infected mice and the controls; (3) CoxB3m virus in myocardial tissue was detected using in situ RT-PCR by direct incorporated technique which employed nucleotide labeling by anti-digoxin antibody and bonded with alkaline phosphatase (anti-dig-AKP method). Results　Picture analysis indicated that the left ventricular chamber area was enlarged and the left ventricular wall was thinner in the chronic repeated virus infected models than those of the controls. With in situ RT-PCR, positive signals for Coxsackie virus B3m RNA were detected not only in the myocardium of the acute Balb/c mice models but also in the myocardium of the chronic mice models. Conclusion　Coxsackie virus B3m is able to induce pathologic lesions by exhibiting positive CVB-RNA signals in both acute and chronic models in mice. In the chronic experimental models, the cardiac chamber is enlarged while the ventricular wall is thinned which demonstrates the association with persistent infection of Coxsackie virus B3m virus.

Objective:To explore the morbidity features and therapeutic outcomes of rejections in pediatric kidney transplantation (KT) recipients.Methods:Between January 2013 and June 2022, 360 children undergoing KT were recruited.The relevant clinical data were collected for examining the morbidity features and therapeutic outcomes of rejections.The serum levels of creatinine were compared among groups by non-parametric rank test.And Kaplan-Meier and Log-rank methods were employed for examining the incidence of rejection and comparing mortality-censored graft survival rates among patients with different times of rejection.Results:A total of 58 recipients had 82 incidents of rejection with a cumulative incidence of 6.3%, 9.2% and 11.3% at 3/6/12 months respectively.Among 50 incidents of biopsy-proved rejections, the types were T cell-mediated rejection [TCMR, 42.0%(21/50)], antibody-mediated rejection [20.0%(10/50), ABMR] and mixed rejection [38.0%(19/50)].Among 58 incidents of initial rejection, 69% had maintained graft function (MGF) and 31% impaired graft function (IGF) after anti-rejection regimens.Among 80.8%, 85.7% and 75% of recipients with clinical rejection, ABMR or borderline rejection while 36.4% in TCMR patients had MGF.Fifteen kidney allografts lost function in 58 recipients with rejection.Five-year death-censored graft survival was significantly lower in patients with two or more incidents of rejection (30.5%, 95% CI: 12.3%-75.4%) than in those without rejection (92.9%, 95% CI: 89.3%-96.6%) ( P<0.000 1) or with only one rejection (82.9%, 95% CI: 65.9%-100%)( P<0.001). Conclusions:The rejection rate remains high in KT children and it affects graft survival.And TCMR is more likely to cause impaired graft function.Recurrent rejections have a more pronounced impact upon graft survival.

Objective:To explore the clinical characteristics and outcomes of pediatric kidney transplantations at a single center and discuss the related clinical issues.Methods:From January 1990 to October 2019, clinical data were analyzed retrospectively for 244 pediatric renal transplants. The youngest recipient was aged 1.8 years and the median age of pediatric recipients was 12.2 years. The major disease was primary or hereditary glomerulonephritis ( n=160, 69.0%), congenital anomalies of kidney and urinary tract (CAKUT), cystic renopathy and other hereditary nephropathies ( n=55, 23.7%). The donor sources included traditional deceased donor ( n=42, 17.2%), living-related donor ( n=19, 7.8%) and organ donation ( n=183, 75.0%). The median age of donors was 2 years (0-51) and the median weight 12.0(2.7-72.0) kg. From January 2013 to October 2019, 170 cases), the major induction immunosuppression regimen was anti-thymocyte globulin (ATG) ( n=110, 64.7%) or basiliximab ( n=58, 34.1%). The maintenance regimen was tacrolimus + mycophenolic acid (MPA) + glucocorticosteroids. Finally the outcomes and the complications were analyzed. Results:The survival rates of 244 kidney allograft recipients were 98.1%, 94.5% and 93.4% and the graft survival rates 92.6%, 84.2% and 82.0% at 1/3/5 years respectively. Ten recipients died of accident ( n=2, 20.0%), pneumonia after transplantation ( n=2, 20.0%) and intracranial hemorrhage ( n=2, 20.0%). Thirty-three recipients lost their allografts mainly due to intravascular thrombosis in graft ( n=5, 14.3%), acute rejection ( n=5, 14.3%) and death ( n=9, 25.7%). Besides, among 109 deceased donor allograft recipients, the postoperative outcomes were delayed graft function recovery (DGF) ( n=27, 24.8%), arterial thrombosis ( n=6, 5.5%), venous thrombosis ( n=1, 0.9%), graft perirenal hematoma ( n=6, 5.5%), raft artery stenosis ( n=10, 9.2%) and graft ureteral fistula ( n=1, 0.9%). The incidence of acute rejection was 17.5% and 23.2% at 1/3 year respectively. The recurrent rate of primary disease was 6.9%, including primary FSGS ( n=3, 42.9%) and IgA nephropathy ( n=2, 28.6%). At 1/3 year post-operation, the incidence of pulmonary infection was 16.9% and 22.4% and the incidence of urinary tract infection 26.9% and 31.7%. Excluding recipients with graft failure, the estimated glomerular filtration rate (eGFR) at 1/2/3 year postoperatively was (80.3±25.2), (81.4±27.8) and (71.8±27.6) ml/(min·1.73 m 2)respectively. Conclusions:The outcomes of pediatric renal transplantations are excellent at our center. Future efforts shall be devoted to optimizing the strategies of donor kidney selection and strengthening preoperative evaluations, perioperative and postoperative managements for improving the long-term outcomes of pediatric renal transplantations.