[Whole exome sequencing analysis and prenatal diagnosis in children with neurodevelopmental disorders].

To explore the application value of whole exome sequencing (WES) in the diagnosis of prenatal and postnatal neurodevelopmental disorders (NDDs). A total of 70 patients diagnosed with NDDs who underwent WES at the Medical Genetics Center of the Maternal and Child Health Hospital of Hubei Province between June 2020 and July 2021 were retrospectively analyzed. Genomic DNA was extracted from peripheral blood samples and amniotic fluid. WES-based copy number variant (CNV) analysis was integrated into the routine WES data analysis pipeline. The results showed that a molecular diagnosis rate could be made in 21/70 (30%) cases. Of 21 positive cases, 14 (23%) cases were detected by single-nucleotide variant/small insertion/deletion (SNV/Indel) analysis, of which 12 variants were novel, 6 (9.8%) cases were detected by WES-based CNV analysis, and 1 (1.6%) case was detected by a combination of both. The diagnostic yield of WES combined with CNV analysis was higher than that of SNV/Indel analysis alone (30%, 21/70 vs. 20%, 14/70). Of the 28 prenatally diagnosed cases, 6 cases were found to have inherited parental variation for NDDs, 10 cases were found not to have the same pathogenic variation as the proband, and the remaining 12 cases were found to have no pathogenic or likely pathogenic variation that could explain the NDDs phenotype. Clinical follow-up showed that 5 families opted for abortion and the remaining had no current abnormalities. In conclusion, WES may be an effective method to clarify the genetic etiology and prenatal diagnosis of NDDs, which is helpful in assessing the prognosis to aid clinical management and reproductive guidance.

[Application and clinical efficacy of ultrasound debridement method in residual burn wounds].

Objective: To investigate the application and clinical efficacy of ultrasound debridement method in residual burn wounds. Methods: A retrospective cohort study was conducted. From August 2017 to August 2021, 64 patients with residual burn wounds who met the inclusion criteria were admitted to the 980th Hospital of the Joint Logistic Support Force of PLA. According to the debridement method adopted for the residual wounds, the patients were divided into ultrasound debridement group (34 cases, 22 males and 12 females, aged (31±13) years) and traditional debridement group (30 cases, 19 males and 11 females, aged (32±13) years). After the corresponding debridement, the wounds of patients in the two groups were selected for stamp skin grafting or large skin grafting according to the wound site and skin donor status. For unhealed wounds after stage Ⅰ surgery, secondary debridement and skin grafting were be performed, with the wound debridement methods in the 2 groups being the same as those of stage Ⅰ, respectively. On postoperative day 3, drug-sensitive test was used to detect the bacteria in the wound and the positive rate of bacteria was calculate. On postoperative day 7, the survival rate of skin slices in wound and the incidence of subcutaneous hematoma were calculated. At discharge, wound healing time and debridement times of patients were counted, and the secondary debridement rate was calculated. Data were statistically analyzed with independent sample t test or chi-square test. Results: On postoperative day 3, the wounds in ultrasound debridement group were infected with Staphylococcus aureus in 2 cases and Pseudomonas aeruginosa in 2 cases, and the wounds in traditional debridement group were infected with Staphylococcus aureus in 5 cases, Pseudomonas aeruginosa in 3 cases, Acinetobacter baumannii in 1 cases, Klebsiella pneumoniae in 1 cases, and Enterobacter cloacae in 1 cases. The positive rate of bacteria of wound in ultrasound debridement group was significantly lower than that in traditional debridement group (χ2=5.51, P<0.05). On postoperative day 7, the survival rate of skin grafts in ultrasound debridement group was (92±5) %, which was significantly higher than (84±10) % in traditional debridement group (χ2=6.78, P<0.01); the incidence of subcutaneous hematoma in ultrasound debridement group was 17.6% (6/34), which was significantly lower than 40.0%( 12/30) in traditional debridement group, χ2=3.94, P<0.05. At discharge, the wound healing time in ultrasound debridement group was (11.0±2.0) d, which was significantly shorter than (13.0±3.1) d in traditional debridement group (t=3.81, P<0.01); the secondary debridement rate of wounds in ultrasound debridement group was 2.9% (1/34), which was significantly lower than 20.0% (6/30) in traditional debridement group (χ2=4.76, P<0.05). Conclusions: Ultrasound debridement method can significantly reduce the bacterial load of residual burn wounds, reduce postoperative hematoma formation, and promote the survival of skin grafts to shorten the course of disease of patients.

[Clinicopathological features and molecular characteristics of tall cell and hobnail variants of papillary thyroid carcinoma].

Objective: To investigate the clinicopathological and molecular genetic characteristics of tall cell variant and hobnail variant of papillary thyroid carcinoma (PTC). Methods: Twenty-one cases of tall cell variant (TCV-PTC) of PTC (TCV-PTC) and ten cases of hobnail variant of PTC (HV-PTC), as the highly aggressive group, were collected from Xuanwu Hospital from August 2009 to August 2015. Twenty-two cases of follicular variant and 21 classical PTC cases were included as control. Relevant clinical and pathologic data were obtained, and in some cases, paraffin samples were selected for gene mutation spectrum analysis using second generation sequencing. Results: There were 18 males and 56 females; 57 patients were younger than 55 years of age, and 17 patients were 55 years or older. The mean tumor size was 1.6 cm for the high-aggressive group (TCV-PTC and HV-PTC), 1.1 cm for the follicular subtype, and 1.6 cm for the classical type. There were 54 cases with thyroid capsule invasion, 24 cases with extra-thyroidal invasion, and 45 patients with lymph node metastases. Regional recurrence occurred in 7 cases, no recurrence in 54 cases, and 13 patients were lost to follow-up. The highly aggressive group was more likely to show extra-thyroidal invasion, lymph node metastases and recurrence than those with classical PTC (P<0.05). Within this cohort, BRAF V600E mutation was detected in 53 cases and TERT promoter mutation in 6 cases. Compared with the single mutation group and no mutation group, BRAF and TERT promoter co-mutation group was more commonly detected in older age, male, larger tumor size and more prone to extra-thyroid invasion (P<0.05). In addition, among BRAF and TERT co-mutation cases, the highly-aggressive group accounted for the highest proportion (5/6). Conclusions: TCV-PTC and HV-PTC, as highly-aggressive variants of PTC, show more aggressive biologic behavior (more lymph node metastasis, external thyroid invasion and recurrences) than the classical and follicular variants of PTC. Coexisting BRAF and TERT promoter mutations may be associated with invasive biologic behavior.

[Clinicopathological and molecular genetic characteristics of adult IDH wild-type diffuse gliomas].

Objective: To analyze the clinicopathological and molecular features and prognostic implications of adult isocitrate dehydrogenase wild type (IDH-wt) diffuse gliomas. Methods: A total of 87 cases of adult IDH-wt diffuse gliomas from 2016 to 2020 in Xuanwu Hospital of Capital Medical University were retrospectively collected. The clinicopathological characteristics and prognosis were analyzed. Molecular characteristics were also analyzed using Sanger sequencing and next generation sequencing. Results: There were 53 males and 34 females, aged from 19 to 78 years (mean 53 years). Histopathologically, there were 63 (72.4%) glioblastomas, 16 (18.4%) anaplastic astrocytomas, six (6.9%) diffuse astrocytomas, and one (1.1%) each of anaplastic oligodendrocytoma, and anaplastic oligodendroglioma. Common molecular genetic changes in IDH-wt gliomas included TERT promoter mutation which was found in 60 cases (69.0%); MGMT promoter methylation in 43 cases (49.4%); EGFR mutation in 38 cases (43.7%); PTEN mutation in 35 cases (40.2%) and TP53 mutation in 32 cases (36.8%). In addition, PDGFRA mutation was detected in 17 cases (19.5%), CDK4 amplification in 15 cases (17.2%) and MDM2 amplification in 11 cases (12.6%). In IDH-wt diffuse gliomas, there was no significant difference in the overall survival between TERT promoter, EGFR, PTEN, TP53, PDGFRA, CDK4, MDM2 mutations and the wild-type, since these gene mutations could co-occur in any case (P>0.05). Also there was no significant difference in the overall survival between the WHO grade Ⅱ/Ⅲ gliomas and glioblastoma patients with these gene mutations (P>0.05). Conclusions: TERT promoter, EGFR, PTEN, TP53, PDGFRA, CDK4 and MDM2 gene mutations are common molecular genetic changes in adult IDH-wt gliomas, and are associated with poor prognosis. It is suggested that these genes are potentially useful for predicting the prognosis and should be tested in adult IDH-wt gliomas.

[Chordoid glioma: a clinicopathological study].

Objective: To analyze the clinicopathological features of chordoid glioma. Methods: A total of 12 cases of chordoid gliomas from 2009 to 2020 in Xuanwu Hospital of Capital Medical University and General Hospital of Chinese People's Liberation Army were retrospectively analyzed. The clinical and imaging characteristics, pathologic and molecular characteristics were analyzed, and the relevant literature was reviewed. Results: All 12 patients (4 males and 8 females) aged from 25 to 67 years (mean 39 years) and mainly had a history of headache or/and vision loss. MRI showed that the lesions located in the third ventricle, and they showed abnormal enhancement. Pathologically, these 12 cases displayed the morphologic characteristics of chordoid gliomas, including papillary structures in two cases. Immunohistochemically, GFAP and vimentin were expressed in all 12 cases (12/12). TTF1 was also expressed in all cases (10/10). CD34 and CKpan were seen in 11 cases (11/12). EMA with dot-and/or-ring like positivity was seen in 9 cases (9/10). Tissues were available in nine chordoid gliomas for Sanger sequencing to detect PRKCA and IDH gene mutation, and eight cases (8/9) showed PRKCA gene D463H mutation. None of these cases showed IDH1 R132 and IDH2 R172 mutation. All 12 patients underwent surgery, and four were lost to follow up. The remaining eight patients were progression or recurrence free at last follow-up in January 2021. Conclusions: Chordoid gliomas have relatively distinguishing clinical and histopathological features. PRKCA gene mutation in chordoid gliomas can be considered as a biomarker for the diagnosis and differential diagnosis of chordoid gliomas, and may provide a direction for future targeted therapy.

[Spatio-temporal patterns of Oncomelania hupensis snail habitats in Suzhou, Wuxi and Changzhou cities along the Taihu Lake region].

OBJECTIVE: To investigate the spatio-temporal distribution characteristics of Oncomelania hupensis snail habitats in three cities of Suzhou, Wuxi and Changzhou along the Taihu Lake region, so as to provide technical supports for establishing a sensitive and highly effective surveillance and forecast system for schistosomiasis. METHODS: Snail distribution data were collected from Suzhou, Wuxi and Changzhou cities from 1950 to 2018, and the changing trend for snail habitats were described over years. In addition, the clusters of snail habitats were detected using Kernel density analysis and SaTScan space-time scan analysis. RESULTS: The number of snail habitats appeared a single-peak distribution in Suzhou, Wuxi and Changzhou cities from 1950 to 2018, which peaked in 1970 and then declined rapidly. There were 62.68% of snail habitats eliminated within 10 years after identification, of which 38.24% were eliminated at the year of identification. Kernel density analysis and SaTScan space-time scan analysis revealed that high-density clusters of snail habitats were mainly distributed in Kunshan City, Wuzhong District and Xiangcheng District from 1970 to 1980, and in Yixing City in 1990; since then, the clusters gradually shrank, and overall appeared a move from northeast to west of Taihu Lake. A total of 4 new clusters were detected after 1970, as revealed by space-time scanning of snail habitats. In current snail habitats, emerging snail habitats are mainly identified in Huqiu District (Dongzhu Town), Wuzhong District (Guangfu Town), Taicang City (Shaxi Town) and Jintan District, and re-emerging snail habitats are scattered in 7 districts. CONCLUSIONS: The distribution of snail habitats are spatio-temporal aggregation in Suzhou, Wuxi and Changzhou cities. The monitoring and prediction of emerging and re-emerging snail habitats are the key points in the future.

The relevance of HIF1A gene polymorphisms and primary hypertensive left ventricular hypertrophy in Chinese Han population.

OBJECTIVE: To investigate the effect of gene polymorphisms in hypoxia-inducible factor 1α (HIF-1α) in left ventricular hypertrophy of hypertensive patients. PATIENTS AND METHODS: A total of 583 hypertensive patients were divided into two groups, with left ventricular hypertrophy (LVH (+), 198 cases) and without left ventricular hypertrophy (LVH (-), 385 cases). Polymerase Chain Reaction restriction fragment length polymorphism was used to detect the single nucleotide gene polymorphisms rs11549465, rs11549467 and rs1957757 in HIF-1α. RESULTS: The distribution differences of gene frequencies for rs11549465, rs11549467 and rs1957757 in HIF1A single nucleotide gene polymorphisms for LVH (+) and LVH (-) were statistically significant (p<0.05). The T allele of rs11549465 loci and the G allele of rs11549467 loci increased the risk of LVH, related to the increased plasma expression of HIF-1α (p<0.05). CONCLUSIONS: Gene polymorphism of HIF-1α were related to primary hypertensive left ventricular hypertrophy, and the expression of rs11549467 correlated with the increasing concentration of plasma HIF-1α.

[Progress of interruption of schistosomiasis transmission in Jiangsu Province].

Schistosomiasis was once heavily endemic in Jiangsu Province. Following the control efforts for several decades, schistosomiasis was almost eradicated in all endemic counties in Jiangsu Province in 1980, and transmission control was achieved in the province in 2011. According to the principle of "implementing the control measures with adaptation to local circumstances and guiding the control programs with classified interventions", an integrated strategy with emphasis on the management of both infectious sources and snails has been recently employed for schitsosomiasis control in Jiangsu Province. In addition, a sensitive and highly effective surveillance system has been built and the application of novel techniques and information construction has been intensified to effectively interrupt the transmission of schistosomiasis in the Province. Transmission interruption of schistosomiasis was achieved in all endemic counties in Jiangsu Province. The paper summarizes the endemic situation of schistosomiasis, progress of schistosomiasis control, and major schistosomiasis control measures implemented during the stage of transmission interruption in Jiangsu Province.

[Endemic situation of schistosomiasis in national surveillance sites of Jiangsu Province from 2011 to 2018].

OBJECTIVE: To analyze and investigate the changing trend of the endemic situation of schistosomiasis in national surveillance sites of Jiangsu Province from 2011 to 2018, so as to provide scientific evidence for formulating strategies for schistosomiasis control. METHODS: From 2011 to 2014, the national schistosomiasis surveillance sites were set in seven schistosomiasis endemic counties (cities, districts) across Jiangsu Province as according to the National Schistosomiasis Surveillance Scheme (2011 version), and from 2015 to 2018, the national surveillance sites were assigned in all 64 counties (cities, districts) endemic for schistosomiasis in Jiangsu Province according to the National Schistosomiasis Surveillance Scheme (2014 version). Schistosoma japonicum infections in local populations, mobile populations and livestock, and snail status were monitored in the national schistosomiasis surveillance sites of Jiangsu Province from 2011 to 2018, and the monitoring data were statistically analyzed. RESULTS: The sero-prevalence of S. japonicum infections was 1.50% to 4.61% among local populations in the national schistosomiasis surveillance sites of Jiangsu Province from 2011 to 2018, and a higher sero-prevalence was seen in men than in women, with the sero-positives predominantly detected in local populations at ages of over 50 years. The positive rate of stool examinations was 0 to 0.14% among sero-positive local populations in the national schistosomiasis surveillance sites of Jiangsu Province from 2011 to 2018, and no acute case was found in local populations during the study period. The sero-prevalence of S. japonicum infections was 0.46% to 15.97% among mobile populations in the national schistosomiasis surveillance sites of Jiangsu Province from 2011 to 2018, and no egg-positives were identified. A total of 1 453 livestock were tested in the national schistosomiasis surveillance sites of Jiangsu Province from 2011 to 2018, and no S. japonicum infections were detected. During the period from 2011 through 2018, snail survey was conducted in an area of 216 million m2 in the national schistosomiasis surveillance sites of Jiangsu Province, and 1 291.01 hm2 snail habitats were identified, with snail densities ranging from 0.01 to 0.47 snails/0.1 m2; however, no S. japonicum infections were identified in snails. CONCLUSIONS: The overall endemic situation of schistosomiasis appears a tendency towards a decline in Jiangsu Province, and S. japonicum infection remains at a low level in both humans and livestock. No S. japonicum infection has been identified in local populations in Jiangsu Province since 2012. In the future, monitoring and management of imported sources of S. japonicum infections should be intensified in Jiangsu Province, and the capability building of passive surveillance of schistosomiasis should be improved in sentinel hospitals in national schistosomiasis surveillance sites of Jiangsu Province. In addition, the examination of schistosomiasis should be strengthened in mobile populations in Jiangsu Province, a sensitive and effective surveillance-response system for schistosomiasis is urgently needed.

[Assessing the efficiency of schistosomiasis control programs in Jiangsu Province from 2005 to 2015 using data envelopment analysis (Malmquist index method)].

OBJECTIVE: To assess the total factor productivity (TFP) of schistosomiasis control programs in Jiangsu Province, so as to provide insights into sustainable schistosomiasis control. METHODS: The data envelopment analysis-Malmquist index method was employed to analyze the human resources and financial investments in schistosomiasis control programs from health sectors in each schistosomiasis-endemic city of Jiangsu Province from 2005 to 2015, and assess the outputs of each schistosomiasis control project. RESULTS: The overall productive efficiency of schistosomiasis control programs in Jiangsu Province showed an increasing tendency, and the mean fluctuation of annual TFP was 2.3%. The comprehensive technical efficiency, including pure efficiency and scale efficiency, appeared a steady increase with minor fluctuations, and the mean fluctuation of annual comprehensive technical efficiency was 3.8%. The growth rate of technical progress fluctuated greatly from 2005 to 2011, and showed a steady increase from 2012 to 2015, which became a major contributor to the growth of TFP. A higher growth rate of TFP was seen in Huai ' an and Changzhou cities, which showed a greater comprehensive technical efficiency, and a large fluctuation was observed in the growth rate of technical progress in Yancheng, Nanjing, Huai ' an and Yangzhou cities. CONCLUSIONS: There is a continuous improvement in the technical level of schistosomiasis control programs in Jiangsu Province, and technical application and supervision and management capacity also show a steady increase. In addition, the application of new techniques and new strategies contributes greatly to TFP growth. In the future, the investment into new techniques and new strategies should be increased to ensure the sustainable schistosomiasis control in Jiangsu Province.

[Application value of a dynamic automatic identification system in Schistosoma japonicum miracidium hatching test].

OBJECTIVE: To evaluate the value of a dynamic automatic identification system in routine miracidium hatching test with nylon gauzes. METHODS: Different quantities of fresh Schistosoma japonicum eggs were added to bovine fecal samples and divided into the low-infection group, medium-infection group and high-infection group, while the bovine feces without S. japonicum eggs served as negative controls. The detection efficiency and accuracy were compared between the identification system and manual detection in different groups. RESULTS: The identification system can automatically identify S. japonicum miracidium. The detection rate and efficiency of S. japonicum miracidium in bovine fecal samples were both higher by using the identification system than by manual detection. Notably in the low-infection group, the identification system had a significantly higher rate of detection of S. japonicum miracidium than manual detection (χ2 = 10.769, P = 0.002). The identification system completed the detection of bovine fecal samples in the field within 1 min. CONCLUSIONS: The dynamic automatic identification system may effectively improve the detection efficiency and accuracy of routine miracidium hatching test with nylon gauzes, and it may replace manual detection to be used in the field schisotsomiasis examinations and related researches.

[Establishment and operation of schistosomiasis diagnostic reference laboratory in Jiangsu Province].

OBJECTIVE: To construct the schistosomiasis diagnostic reference laboratory in Jiangsu Province, and to examine the role and diagnostic efficiency of the reference laboratory. METHODS: A schistosomiasis diagnostic reference laboratory was built in Jiangsu Province according to the requirements of the construction of the national schistosomiasis diagnostic reference laboratory in China. Inter-laboratory comparisons were conducted and the diagnostic capability of grassroots laboratories was evaluated in Jiangsu Province. RESULTS: The organization structure, environmental conditions, administration and quality systems of the schistosomiasis diagnostic reference laboratory in Jiangsu Province all met the requirements for construction of the national schistosomiasis diagnostic reference laboratory in China, and the schistosomiasis diagnostic reference laboratory in Jiangsu Province was issued a certificate of a province-level schistosomiasis diagnostic reference laboratory. During the 6 inter-laboratory comparisons performed by national schistosomiasis diagnostic reference centers of China, the qualitative and quantitative results of each detection item were all in agreement with the reference samples (Kappa = 1), and the diagnostic capability was identified excellent. The results of indirect hemagglutination assay of 426 serum samples from 4 grassroots laboratories were re-examined, and the mean coincidence rate was 94.13% (range, 92.08% to 96.25%) with the grassroots laboratories, with a mean Kappa value of 0.85 (range, 0.83 to 0.86) and a mean missing rate of 10.19% (range, 0 to 17.65%). CONCLUSIONS: The schistosomiasis diagnostic reference laboratory has been successfully established and effectively operated in Jiangsu Province, which plays an active role in improving the capability of schistosomiasis diagnostic equality in the province.

A new cell culture system for infection with hepatitis B virus that fuses HepG2 cells with primary human hepatocytes.

Hepatitis B virus (HBV) infection exhibits a very narrow host range and shows a strong tropism for liver parenchymal cells, however none of the previously established experimental models can reproduce the natural process of HBV infection. In the present study, primary human hepatocytes were fused with HepG2 cells to establish the hybrid HepCHLine-4 cell line with high susceptibility to HBV. The HepCHLine-4 cells expressed HBV-specific antigen when co-incubated with HBV-positive serum from a hepatitis B patient. Post-infection, HBV relaxed circular DNA and covalently closed circular DNA were detected in HepCHLine-4 cells using a nested polymerase chain reaction, and HBV-specific particles were visualized by electron microscopy of the culture media of HepCHLine-4 cells. HepG2 cells were not susceptible to HBV infection under the same conditions. The HepCHLine-4 cells can be sub-cultured for > 12 months while maintaining susceptibility to HBV and may, therefore, be useful for studying HBV infection and the viral life cycle in human hepatocytes.

Tissue factors on acute promyelocytic leukemia and endothelial cells are differently regulated by retinoic acid, arsenic trioxide and chemotherapeutic agents.

The aberrant expression of tissue factor (TF) in acute promyelocytic leukemia (APL) cells has been implicated in the pathogenesis of the APL coagulopathy. In this study, we found that in APL patients receiving ATRA or As2O3 treatment, the improvement in hypercoagulobility and hyperfibrinolysis paralleled the correction of plasma fibrinogen level and amelioration of bleeding symptoms. Notably, clinical improvement was also correlated to ATRA/As2O3-induced rapid decrease of membrane procoagulant activity (PCA) and TF contents of APL blasts. Consistent with the in vivo findings, the membrane PCA, TF antigen and its mRNA level within NB4 cells were rapidly down-regulated by 1 microM ATRA or As2O3, while 0.2 microg/ml DNR increased these TF parameters prior to its effect upon apoptosis induction. The down-regulation of TF mRNA by ATRA was partially de novo protein synthesis-dependent and at least partially attributed to a mechanism of destabilizing TF mRNA. On the other hand, in addition to its modulation on mRNA, As2O3 could also induce an accelerated TF protein turnover. These distinct effects were corroborated with the properties of these agents in causing the degradation of PML-RARalpha protein. All three therapeutic agents, however, enhanced the potential of NB4 cells to stimulate the expression of TF and PCA in endothelium. Taken together, our data suggest that the rapid and distinct regulation of TF on APL cells by these therapeutic agents might at least partially contribute to their effects on APL coagulopathy.

Effects of indomethacin on joint damage in rat and rabbit.

AIM: To study the effects of indometacin (Ind) on joint damages. METHODS: The volume of noninjected hind paw and interleukin-1 (IL-1) production from peritoneal macrophages and articular synoviocytes induced by lipopolysaccharides were assayed in adjuvant arthritis (AA) rats. Measurements of synovial fibroblast proliferative response and proteoglycan synthesis of cartilage from rabbits were used. RESULTS: The secondary inflammatory reactions in AA rats on d 18, 21, and 24 were suppressed by i.g. Ind 2 mg.kg-1.d-1 for 9 d. Ind promoted IL-1 production from both macrophages and synoviocytes in AA rats. Ind 10 mumol.L-1 enhanced the proliferation of rabbit synovial fibroblasts and suppressed the proteoglycan synthesis of articular cartilage in response to IL-1 in vitro. CONCLUSION: Ind is unfavorable to the repair of joint destruction.