Validation and analysis of Goldengate high-throughput deafness gene chip in detecting the patients with enlarged vestibular aqueduct syndrome / 中华耳鼻咽喉头颈外科杂志

Objective: To verify the accuracy and effectiveness of Goldengate high-throughput deafness gene chip in detecting the patients with enlarged vestibular aqueduct syndrome(EVAS), and to provide a reference for genetic detection strategy of EVAS. Methods: From August 2016 to February 2018, 15 patients with EVAS and 60 normal controls were detected by Goldengate high-throughput deafness detection chip developed by our team, and the results were verified by Sanger sequencing. SLC26A4 gene sequencing was carried out in all the patients with EVAS. Results: 12/15 of patients with EVAS were detected mutations of SLC26A4 gene. Nine mutations were detected by chip detection and SLC26A4 gene direct sequencing, seven of which were detected by both methods. The chip could detect 93.33%(28/30) of the allele information provided by SLC26A4 gene direct sequencing. In addition to SLC26A4 gene, mutations of GJB2, PCDH15, TMC1, MYO6 and mitochondrial genes were detected in 15 patients with EVAS. These results were verified by Sanger sequencing. Conclusion: Goldengate high-throughput deafness gene chip possesses the traits of wide coverage and high accuracy, which can be used as a preliminary detection method for patients with EVAS.

Alterations of Thymic Epithelial Cells in Lipopolysaccharide-induced Neonatal Thymus Involution / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Vascular endothelial growth factor (VEGF) in the thymus was mainly produced by the thymic epithelial cells (TECs), the predominant component of the thymic microenvironment. The progression of TECs and the roles of VEGF in the neonatal thymus during sepsis have not been reported. This study aimed to explore the alterations of TECs and VEGF level in the neonatal thymus involution and to explore the possible mechanisms at the cellular level.</p><p><b>METHODS</b>By establishing a model of clinical sepsis, the changes of TECs were measured by hematoxylin-eosin staining, confocal microscopy, and flow cytometry. Moreover, the levels of VEGF in serum and thymus were assessed based on enzyme-linked immunosorbent assay and Western blotting.</p><p><b>RESULTS</b>The number of thymocytes and TECs was significantly decreased 24 h after lipopolysaccharide (LPS) challenge, (2.40 ± 0.46)×10 7 vs. (3.93 ± 0.66)×10 7 and (1.16 ± 0.14)×10 5 vs. (2.20 ± 0.19)×10 5 , P < 0.05, respectively. Cortical TECs and medullary TECs in the LPS-treated mice were decreased 1.5-fold and 3.9-fold, P < 0.05, respectively, lower than those in the controls. The number of thymic epithelial progenitors was also decreased. VEGF expression in TECs was down-regulated in a time-dependent manner.</p><p><b>CONCLUSION</b>VEGF in thymic cells subsets might contribute to the development of TECs in neonatal sepsis.</p>

Screening of differentially expressed microRNAs in borderline and malignant gastrointestinal stromal tumors / 中华病理学杂志

<p><b>OBJECTIVE</b>Gastrointestinal stromal tumors (GISTs) have a broad spectrum of biological behaviors ranging from benign, borderline and malignant. This study aimed to screen differentially expressed microRNAs (miRNAs) between malignant and borderline GISTs and to investigate the potential role of miRNAs in the malignant transformation of GISTs.</p><p><b>METHODS</b>Six GIST samples including borderline tumors (n = 3) and malignant tumors (n = 3) were collected based on the clinical and pathological characteristics. Total RNA was extracted, followed by miRNA microarray analysis to screen the differentially expressed miRNAs. The most significantly expressed 4 miRNAs were then chosen for further validation by real-time PCR in 22 additional GIST samples.</p><p><b>RESULTS</b>Direct comparison of malignant group versus borderline group revealed 14 significantly and differentially expressed miRNAs (P < 0.05, with a fold change of < 0.5 or > 2). Five miRNAs were up-regulated and nine were down-regulated in the malignant group. Four miRNAs (miR-221, miR-135b, miR-675(*) and miR-218) were most significantly and differentially expressed between the two groups. The differential expression of 2 miRNAs (miR-221 and miR-675(*)) were subsequently confirmed with good concordance by real-time PCR.</p><p><b>CONCLUSIONS</b>The differential miRNA expression profiles between two groups are revealed by miRNA microarray assay, and confirmed by real-time PCR. Among differentially expressed miRNAs, miR-221 and miR-675(*) might be related to the malignant transformation of GISTs, and have a potential value in predicting biological behavior of GISTs.</p>

Detection of KRAS gene mutation and its clinical significance in colorectal adenocarcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the clinical significance of KRAS mutation detection in colorectal adenocarcinoma.</p><p><b>METHODS</b>Paraffin-embedded tissue specimens were obtained from 440 patients with colorectal adenocarcinoma. The genomic DNA was extracted. Mutations of exon 2 of KRAS gene were examined by PCR and direct sequencing.</p><p><b>RESULTS</b>Somatic mutations of KRAS gene were identified in 146 cases, with the mutation rate of 33.2% (146/440). Among these 146 patients, KRAS mutation involved codon 12 in 118 patients, including 35G > A (Gly12Asp, 62 cases), 35G > T (Gly12Val, 35 cases), 34G > T (Gly12Cys, 9 cases), 34G > A (Gly12Ser, 6 cases), 35G > C (Gly12Ala, 5 cases), and 34G > C (Gly12Arg, 1 case); in 27 patients the mutation involved codon 13, including 38G > A (Gly13Asp, 25 cases), 38G > C (Gly13 Val, 1 case) and 37G > T (Gly13 Cys, 1 case); and in one patient, the mutation involved codon 14 with 40G > A (Val14Ile). The status of KRAS or codon 12 mutations in colorectal adenocarcinoma was related to patients' gender (P = 0.021 and P = 0.030, respectively), and this significant correlation to females was conserved in clinical stage III (P = 0.007 and P = 0.003, respectively), but not in stages I, II, and IV. The status of KRAS or codon 12 mutations was also related to tumor stage. Between stage II and stage IV, the mutation rate of KRAS and codon 12 showed significant difference (P = 0.028 and 0.034, respectively). Between stage III and stage IV, only the codon 12 mutation rate showed significant difference (P = 0.011). Codon 13 mutation was not related to tumor stage.</p><p><b>CONCLUSION</b>About one third of patients with colorectal adenocarcinoma have KRAS gene mutation, which might be related to patients' gender; and could be consistently detected by PCR and direct sequencing.</p>

Staging and histologic grading of gastrointestinal stromal tumors / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the clinical stage and histological grade of gastrointestinal stromal tumors.</p><p><b>METHODS</b>Twelve clinical and pathological parameters were assessed in 613 patients with follow-up information. These parameters were classified into two gross spread parameters including liver metastasis and peritoneal dissemination, five microscopic spread parameters including lymph node metastasis, vascular, fat, nerve and mucosal infiltration, and five histological parameters including mitotic count ≥ 10 per 50 high-power fields, muscularis propria infiltration, coagulative necrosis, perivascular pattern and severe nuclear atypia.</p><p><b>RESULTS</b>The accumulated 5-year disease-free survival (DFS) and overall survival (OS) of 293 patients without any of these predictive parameters of malignancy were 99.3% and 100.0%, respectively. They were regarded as nonmalignant and further evaluations on the stage and grade of these tumors were not performed. At least one and at most seven predictive parameters of malignancy were identified in 320 patients. For these patients, the accumulated 5-year DFS and OS rates were 43.9% (mean 6.7 years) and 59.7% (mean 9.3 years), respectively. The DFS showed significant difference between patients with and without gross spread (P < 0.01), with and without microscopic spread (P = 0.001). DFS and OS were associated with the number of predictive parameters of malignancy in patients without gross spread (P < 0.01 for both DFS and OS), but not in patients with gross spread (P = 0.882 and 0.441, respectively).</p><p><b>CONCLUSIONS</b>Malignant GIST could be divided into clinical stages I and II based on the absence and presence of gross spread, respectively. The degree of malignancy of patients in clinical stage I could be graded according to the number of predictive parameters of malignancy. Patients in clinical stage II were of the highest degree of malignancy regardless of the number of parameters. The staging and grading of gastrointestinal stromal tumors in this study are strongly associated with prognosis.</p>

New prognostic parameters for very-low-risk gastrointestinal stromal tumors / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>According to the National Institutes of Health consensus criteria, gastrointestinal stromal tumors (GISTs) smaller than 2 cm in diameter with less than 5 mitotic figures per 50 high-power fields are considered very-low-risk GISTs, but these two indices alone cannot reliably predict a benign outcome during long-term follow-ups. Therefore, identification of additional parameters for predicting the clinical behavior of GISTs is necessary.</p><p><b>METHODS</b>Eighty-eight patients with tumors that meet the very-low-risk GIST criteria were retrospectively investigated and morphological parameters of tumors associated with the biological behavior of very-low-risk GISTs were evaluated in the present study. The Kaplan-Meier method was used to calculate disease-free survival rates.</p><p><b>RESULTS</b>Eighty-one patients were followed up for one to 16.3 years. Five cases of relapses were identified in the patients. Distinctive infiltrative growth patterns such as muscularis propria, muscularis mucosa, or nerve infiltration were identified by microscopy in 4 patients with the relapse, including three patients who experienced multiple recurrences. The infiltrative growth features became more obvious in multiple recurrent tumors compared to the single recurrent tumor, while only one developed relapse in 76 patients without infiltration (P < 0.0001).</p><p><b>CONCLUSION</b>Microscopic infiltrative growth patterns of the tumor may have clinical significance in predicting the prognosis of very-low-risk GISTs.</p>

Comparison of therapeutic effects in severe nocturia: gradual versus immediate drug withdrawal / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate possible differences in the prognosis in children with severe nocturia who received different drug withdrawal schedules.</p><p><b>METHODS</b>Ninety-seven children with severe nocturia were randomly assigned to two groups: control (n=47) and observed (n=50). The control group accepted drug withdrawal immediately, while the observed group accepted dose tapering gradually after a 12-week treatment course. The frequency of enuresis was observed three months after complete drug withdrawal.</p><p><b>RESULTS</b>During the treatment, the frequency of enuresis in all of children from both the control and the observed groups was reduced by over 90%. Forty-six children (92%) from the observed group showed the frequency of enuresis was reduced by over 90%, but 28 children (60%) from the control group (p<0.01) three months after the complete drug withdrawal. There were no significant differences in the adverse effect and the medication compliance between the two groups.</p><p><b>CONCLUSIONS</b>The different schedules of drug withdrawal may lead to different prognosis, and the schedule of gradual drug withdrawal may be superior to the immediate one in children with nocturnal enuresis.</p>

Sclerosing angiomatoid nodular transformation of spleen: a clinicopathologic study of 10 cases with review of literature / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features, differential diagnosis and pathogenesis of sclerosing angiomatoid nodular transformation of spleen.</p><p><b>METHODS</b>Ten cases of sclerosing angiomatoid nodular transformation of spleen were retrieved from the archival file. Histochemical and immunohistochemical (EnVision method) studies were performed. Ultrastructural findings were also available in one of them.</p><p><b>RESULTS</b>Sclerosing angiomatoid nodular transformation was characterized by micronodular appearance of vascular spaces lined by plump endothelial cells with interspersed ovoid spindle cells. Immunohistochemical study showed that the endothelial cells of vessels in the angiomatoid nodules had various expressions of immunologic phenotypes and could be mainly classified into 3 types: CD34(+)/CD31(+)/CD8⁻ endothelial cells of the capillaries, CD8(+)/CD31(+)/CD34⁻ lining cells of the sinusoids and CD31(+)/CD8⁻/CD34⁻ endothelial cells of the small veins. Collagen network and dilated lymphatic sinuses were evident under transmission electron microscope.</p><p><b>CONCLUSIONS</b>Sclerosing angiomatoid nodular transformation of spleen is a rare benign entity. It may represent a reactive condition and bears some relationship with splenic angioma. It needs to be distinguished from borderline or malignant vascular tumors of spleen.</p>

Clinical and pathological studies of borderline gastrointestinal stromal tumors / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Borderline gastrointestinal stromal tumors (GISTs) are intermediate tumors between benign and malignant variants; however, the clinical and pathological features of borderline GISTs remain poorly defined. This study aimed to characterize GISTs and to identify a set of borderline criteria for practical use.</p><p><b>METHODS</b>Medical records and specimens of 840 patients from 12 hospitals were retrospectively examined. Totally 485 and 76 patients with any of the parameters predictive of either malignant or benign tumors were excluded. The Kaplan-Meier method was used to calculate disease-free survival and overall survival rates.</p><p><b>RESULTS</b>Among the remaining 279 borderline GIST patients, 223 were followed up for 1 to 31.48 years. Two patients developed local recurrence, and both were cured by subsequent operations alone. The 5-year disease-free survival and overall survival rates were 99% and 100%, respectively. Morphologically, borderline GISTs typically exhibited moderate cellularity, and subsets of them also showed moderate atypia, low mitotic activities, or large tumor size. According to the National Institutes of Health (NIH) consensus criteria, the risk levels of the 279 GISTs were classified to be very low to high. However, the disease-free survival rates were not significantly different among these risk groups (P = 0.681).</p><p><b>CONCLUSIONS</b>The proposed borderline GIST criteria in the current study may complement the existing NIH criteria, based primarily on tumor size and mitotic count, in the evaluation of the biological behaviors of GISTs. Since a subset of borderline GISTs with high risk level showed favorable outcome, the introduction of the borderline GIST system may avoid overdiagnosis and over therapy.</p>

Functional bladder capacity in 1,500 children with nocturnal enuresis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the association of functional bladder capacity with the severity of bedwetting in children with nocturnal enuresis.</p><p><b>METHODS</b>A questionnaire investigation was performed in 1 500 children with nocturnal enuresis and the functional bladder capacity was examined by B-ultrasound.</p><p><b>RESULTS</b>The ratio of males to females was 1.3:1. The majority of patients (87%) were in an age range of 5-10 years, followed by the 10-14 years group (12%), and the 15-18 years group (1%). Six hundred and thirty-seven patients (42.4%) showed a decreased functional bladder capacity (less than 50% of normal level). The patients were classified into four groups according to the severity of bedwetting (from severe to mild): > or =2 times per night (n=53, 3.5%), > or =7 times per week (n=969, 64.6%), 3-6 times per week (n=380, 25.3%) and 1-2 times per week (n=98, 6.5%). The incidence of the reduction in functional bladder capacity in the above four groups was 79.2%, 48.3%, 29.7% and 14.3% respectively and a significant difference was noted among the four groups.</p><p><b>CONCLUSIONS</b>Most of children with nocturnal enuresis showed decreased functional bladder capacity. Functional bladder capacity is associated with the severity of bedwetting in children with nocturnal enuresis.</p>

Expression and significance of Toll-like receptors in cord blood mononuclear cells / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the expression of TLR4/2 mRNA in neonatal cord blood mononuclear cells (MNC).</p><p><b>METHODS</b>Forty-six neonates without asphyxia and 40 neonates with asphyxia were divided into groups depending on the gestational age. In the neonates without asphyxia, there were 18 full term infants (the gestational age > or = 37 weeks), 16 preterm infants whose gestational age was > or = 32 weeks but < 37 weeks, and 12 preterm infants whose gestational age was < 32 weeks. In the neonates with asphyxia, 11 were full term infants, 15 were preterm infants whose gestational age was > or = 32 weeks but < 37 weeks and 14 were preterm infants at gestational age < 32 weeks. MNCs were separated and cultured with LPS (1 microg/ml) for 3 h. Cells were collected for analysis of gene expression of TLR4/2 by RT-PCR technique. Cell supernatants were taken to measure TNF-alpha production following the ELISA protocol. Fifteen healthy adults were enrolled into the control group. In addition, the Pearson correlation analyses were carried out between the levels of TLR4, TLR2 mRNA and the levels of TNF-alpha.</p><p><b>RESULTS</b>In the neonates without asphyxia, TLR4, TLR2 mRNA and TNF-alpha levels were 0.75 +/- 0.12, 0.63 +/- 0.08, 2502.6 +/- 273.1 ng/L, separately, in the full term infants, 0.37 +/- 0.04, 0.32 +/- 0.03, 1218.8 +/- 145.7 ng/L, separately, in the preterm infants whose gestational ages were > or = 32 weeks but < 37 weeks, and 0.26 +/- 0.03, 0.20 +/- 0.03, 811.8 +/- 105.2 ng/L separately, in the preterm infants whose gestational ages were < 32 weeks. In the neonates with asphyxia, TLR4, TLR2 mRNA and TNF-alpha levels were 0.58 +/- 0.07, 0.50 +/- 0.06, 1946.4 +/- 244.2 ng/L, separately, in the full term infants, 0.29 +/- 0.03, 0.26 +/- 0.03, 970.0 +/- 94.3 ng/L, separately, in the preterm infants whose gestational age was > or = 32 weeks but < 37 weeks, and 0.17 +/- 0.02, 0.14 +/- 0.02, 652.6 +/- 60.3 ng/L, separately, in the preterm infants whose gestational age was < 32 weeks. The levels of TLR4, TLR2 mRNA and TNF-alpha in the adults were 2.71 +/- 0.75, 2.61 +/- 0.33, 9270.1 +/- 1098.3 ng/L, separately. In the preterm infants and full term infants, the levels of TLR4, TLR2 mRNA and TNF-alpha were lower in comparison to the adults. The lower the gestational age, the lower the levels of TLR4, TLR2 mRNA and TNF-alpha. There were significant differences between the levels of TLR4, TLR2 mRNA and TNF-alpha of the neonates without asphyxia and those of the neonates with asphyxia. In the neonates with asphyxia, the levels of TLR4, TLR2 mRNA and TNF-alpha were lower than those in the neonates without asphyxia (P < 0.01). Whether the neonates were asphyxic or not, the levels of TLR4, TLR2 were paralleled with the levels of TNF-alpha.</p><p><b>CONCLUSIONS</b>The expression of TLRs in the neonates, especially in the preterm infants was lower than that in the adults, which probably contributes to the susceptibility of neonates to infections.</p>

Protective effect of N-acetylcysteine against lipopolysaccharide injury in hepatocytes of neonatal mice / 中华儿科杂志

<p><b>OBJECTIVE</b>N-Acetylcysteine (NAC) is a sulfhydryl donor molecule with antioxidant and antiinflammatory effects. A major role has been described for inducible nitric oxide (NO) synthase in several inflammatory liver diseases. NAC attenuates NO generation following lipopolysaccharide injection in rats. The purpose of this study was to investigate the effect of NAC against lipopolysaccharide injury in hepatocytes of neonatal mice and the molecular mechanisms by which NAC influences inflammatory responses of the hepatocytes.</p><p><b>METHODS</b>The liver of neonatal mouse was digested by collagenase to dissociate the hepatocytes. The hepatocytes were cultured and isolated. After 7 days of culture the normal hepatocytes were divided into two groups: LPS group and NAC group. In LPS group, 10 microg/ml LPS was added into the culture medium. In NAC group, 5 mmol/L NAC was added into the culture medium firstly, 10 microg/ml LPS was added after 1 h of culture. There were 12 mice in each group. The cell supernatants and the hepatocytes were collected at 0, 6 and 12 hours after adding LPS. The cell supernatants were taken to measure the alanine aminotransferase (ALT) level and nitric oxide (NO) production by the biochemical methods. The cells were taken to analyze the gene expression of induced nitric oxide synthase (iNOS) by the RT-PCR.</p><p><b>RESULTS</b>In LPS group, the levels of ALT, NO and iNOS mRNA increased significantly at the time points 6 h and 12 h compared with the time point 0, (P < 0.01). Compared with the LPS group, the levels of ALT, NO and iNOS mRNA of NAC group were lower at the time points 6 h and 12 h (P < 0.01).</p><p><b>CONCLUSIONS</b>NAC may play a protective role in the hepatocytes injury caused by LPS in the neonatal mice. The protective mechanism works partially through the inhibition of iNOS activation by LPS.</p>

Studies on the spatial distribution and environmental factors of highly pathogenic avian influenza in Mainland China, using geographic information system technology / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the spatial distribution of highly pathogenic avian influenza (HPAI) and to explore environmental factors associated with HPAI using geographic information system (GIS) techniques in Mainland China.</p><p><b>METHODS</b>Databases were set up using the information of HPAI during epidemics in 2004, and linked to digital maps at provincial and county administrative layers in the country through the ArcGIS 8.3 software. Spatial cluster analyses, spatial statistics analyses and tracking analyses on epidemic situation of HPAI were implemented. Environmental factors associated with HPAI were also analyzed on data related to weather, vegetation and migratory birds etc.</p><p><b>RESULTS</b>Findings from spatial cluster analyses showed that high incidence area was centralized in 113.261 degrees ordm; east longitude and 23. 119 degrees ordm; north latitude with a radius of 1090.52 kilometers (relative risk= 2.646, P value= 0.001). Spatial statistical analyses showed that HPAI took place mainly in capital cities of provinces and surrounding areas as well as in the circumference areas of arterial rivers, lakes and seacoasts. Results also showed that HPAI occurrences were associated with low air temperature, high relative humidity and high air pressure as well as with east & central migration routes of migratory birds. The average normalized difference vegetation index was 0.36 +/- 0.11 in epidemic areas of HPAI.</p><p><b>CONCLUSION</b>HPAI was unrandomly distributed and geographically clustered in China.</p>

Clinical Aspects and Treatment of Enuresis Companied with Spina Bifida Occulta in Children / 实用儿科临床杂志

Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.