Congenital bilateral absence of the vas deferens (CBAVD) is observed in 1%-2% of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator (CFTR) mutations. CFTR is one of the most well-known genes related to male fertility. The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia (NOA). CFTR mutations are highly polymorphic and have established ethnic specificity. Compared with F508Del in Caucasians, the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis. However, whether p.G970D participates in male infertility remains unknown. Herein, a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA. Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation (4.1%, 5/122), excluding polymorphic sites. Furthermore, we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients. The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells. In spermatocyte GC-2(spd)ts (GC2) Cftr p.G965D cells, RNA splicing variants were detected and CFTR expression decreased, which may contribute to the phenotypes associated with impaired spermatogenesis. Thus, this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.
Humans ; Animals ; Mice ; Male ; Mutation, Missense ; Retrospective Studies ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics* ; Infertility, Male/genetics* ; Mutation ; Vas Deferens/abnormalities* ; Spermatogenesis/genetics*

OBJECTIVE: To detect the expression level of HK2 gene in the bone marrow of newly diagnosed patients with acute myeloid leukemia (AML) and investigate its influence on the clinical characteristics and prognosis. METHODS: The expression level of HK2 gene in the bone marrow of 90 newly diagnosed patients with AML that accompanying clinical characteristics and survival status were detected by RT-qPCR, and compared with 18 allogeneic hematopoietic stem cell transplantation (allo-HSCT) donors. The Chi-square test, Kaplan-Meier survival analysis, and Cox proportional hazards regression model were used to analyze the correlation of HK2 expression level with clinical characteristics and prognosis. RESULTS: Compared with allo-HSCT donors, the HK2 expression was significantly increased in newly diagnosed AML patients (P <0.01). Compared with patients with total response (OR, complete response + complete response with incomplete hematologic recovery) after 2 courses of induction chemotherapy, the expression of HK2 in patients without OR was significantly increased (P <0.05). There was a significant difference in the relative expression of HK2 between patients with and without OR after 2 courses of induction therapy (P <0.001). The median survival time of patients with high expression of HK2 was significantly shorter than that of patients with low expression of HK2 (P <0.05). The multivariate Cox proportional hazards regression analysis showed that prognostic stratification, the expression level of HK2, and whether two courses of induction therapy achieved OR were independent factors affecting the prognosis of AML patients (P <0.05). CONCLUSIONS Compared with allo-HSCT donors, the expression level of HK2 gene is increased in the bone marrow of newly diagnosed AML patients. The prognosis of patients with high expression of HK2 is poor. The expression level of HK2 is an independent factor affecting the prognosis of AML patients.
Humans ; Bone Marrow ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Leukemia, Myeloid, Acute/therapy* ; Prognosis ; Retrospective Studies ; Transplantation, Homologous/adverse effects*

OBJECTIVES: To study the biological processes and functions of serum exosomes in children in the acute stage of Kawasaki disease (KD), so as to provide new biomarkers for the early diagnosis of KD. METHODS: In this prospective study, 13 children with KD who were treated in Children's Hospital of Soochow University from June 2019 to August 2020 were enrolled as the KD group, and 13 children who were hospitalized due to bacterial infection during the same period were enrolled as the control group. Whole blood was collected on the next morning after admission, serum samples were obtained by centrifugation, and exosomes were extracted through ultracentrifugation. Serum exosomes were analyzed by label-free quantitative proteomics, and differentially expressed proteins (DEPs) were screened out for functional enrichment analysis. A protein-protein interaction (PPI) network was plotted, and unique proteins were validated by targeted proteomics. RESULTS: A total of 131 DEPs were screened out for the two groups, among which 27 proteins were detected in both groups. There were 48 unique DEPs in the KD group, among which 23 were upregulated and 25 were downregulated, and these proteins acted on "complement and coagulation cascades" and "the MAPK signaling pathway". Validation by targeted proteomics showed that FGG, SERPING1, C1R, C1QA, IGHG4, and C1QC proteins were quantifiable in the KD group. A total of 29 proteins were only expressed in the control group, among which 12 were upregulated and 17 were downregulated. Four proteins were quantifiable based on targeted proteomics, i.e., VWF, ECM1, F13A1, and TTR. A PPI network was plotted for each group. In the KD group, FGG and C1QC had close interaction with other proteins, while in the control group, VWF had close interaction with other proteins. CONCLUSIONS The serum exosomes FGG and C1QC in children in the acute stage of KD are expected to become the biomarkers for the early diagnosis of KD. For children with unexplained fever, detection of FGG, C1QC1, and VWF may help with etiological screening.
Biomarkers ; Child ; Exosomes ; Extracellular Matrix Proteins ; Humans ; Mucocutaneous Lymph Node Syndrome/diagnosis* ; Prospective Studies ; Proteomics ; von Willebrand Factor

Objective: To investigate the clinical features and prognosis in patients with idiopathic sudden sensorineural hearing loss (ISSNHL) with blood-labyrinth barrier breakdown (BLB-B). Methods: Clinical data of patients with unilateral ISSNHL hospitalized from December 2017 to December 2018 were retrospectively analyzed. According to the results of 3D-FLAIR MRI and enhanced MRI scanning, these patients were divided into two groups, i.e., normal and abnormal inner ear groups. The patients in abnormal inner ear group were further divided into two subgroups: BLB-B and BLB-B with exudation. The differences and correlations among the groups in clinical characteristics, in terms of gender, age, deafness side, basic diseases, dizziness/vertigo, vestibular function, hearing loss degree, as well as classification of hearing curve, and prognosis were analyzed by statistical software SPSS 23.0. Results: Data were collected from 150 cases, in which 68 were male and 82 were female, aged (46.2±14.6) years, including 67 cases with normal inner ears and 83 cases with abnormal inner ears (13 cases with BLB-B; 70 cases with BLB-B and exudation). The dizziness/vertigo incidence, side ratio, hearing loss degree, classification of hearing curve, vestibular dysfunction (vestibular double temperature test, HIT and VAT) and therapeutic effect were different between normal and abnormal inner ear groups (P<0.05). The dizziness/vertigo incidence, side ratio, hearing loss degree, classification of hearing curve, vestibular dysfunction (vestibular double temperature test, o/cVEMP, HIT and VAT) and therapeutic effect were different among normal inner ear, BLB-B and BLB-B with exudation groups (P<0.05). Pairwise comparison between groups revealed that vestibular dysfunction (vestibular double temperature test, o/cVEMP, HIT and VAT) and therapeutic effect were different between normal inner ear and BLB-B groups (P<0.05); The dizziness/vertigo incidence, side ratio, hearing loss degree, classification of hearing curve, vestibular dysfunction (vestibular double temperature test, o/cVEMP, HIT and VAT) and therapeutic effect were different between normal inner ear and BLB-B with exudation groups (P<0.05). There was no significant different between BLB-B and BLB-B with exudation groups. Conclusion: BLB-B displayed by 3D-FLAIR MRI manifestation in ISSNHL patients indicates more serious cochlear and vestibular dysfunction, and worse therapeutic effect.
Dizziness ; Female ; Hearing Loss, Sensorineural ; Hearing Loss, Sudden ; Humans ; Male ; Prognosis ; Retrospective Studies ; Vertigo ; Vestibule, Labyrinth

Objective To investigate serum levels of soluble matrix lysin 2 (sST2) in patients with different stages of heart failure and its relationship with prognosis. Methods Data of 300 patients with heart failure of stages A, B, C and D were included in this study. Thirty-three cases of healthy elderly population for physical examination were used as control group. The general information, echocardiography and related biochemical tests containing sST2 and NT-proBNP were collected in the two groups. The survival periods of patients were evaluated according to the Seattle heart failure mode (SHFM). Patients were followed up for 1 year to record the occurrence of adverse events. Results The sST2 level was higher in heart failure group than that of control group. The sST2 level began to increase in stage B, and which increased with the development of cardiac function staging. The sST2 levels were significantly higher in stages B, C and D than those of stage A, and which were significantly higher in stage D than those of stages B and C (P<0.05). There were significantly higher incidence rates of adverse events, left ventricular end diastolic diameter (LVEDD) and left ventricular mass index (LVMI) in the patients with high sST2 level than those of patients with lower sST2 level (P<0.05). Values of sST2, NT-proBNP, LVEDD and LVMI were significantly higher, and values of LVEF and SHFM life expectancy were significantly lower, in patients with adverse events than those of patients without adverse events (P<0.05). There was a negative correlation between sST2 and LVEF, and positive correlation between sST2 with NT-proBNP, LVEDD and LVMI (P<0.05). The size under ROC curve, which was used to predict the cardiovascular endpoint events judged by sST2 was 0.665 (95％CI:0.574-0.757, P<0.01), and the one by NT-proBNP was 0.790 (95％ CI: 0.731-0.848, P<0.01). The best cut-off value of predicting the clinical adverse events was 139.27μg/L by sST2 and 855.35μg/L by NT-proBNP. Conclusion The serum level of sST2 is early indicator of heart failure, which not only reflects the severity of ventricular remodeling but also is one of indicators to estimate the prognosis of heart failure in one year.

OBJECTIVETo study the coagulation properties the refrigerated whole blood stored at 4℃.

METHODSTen units of whole blood were obtained from healthy volunteer donors and stored at 4±2℃ for 21 days. Samples were collected on the day after donation and on days 2, 4, 6, 8, 10, 14 and 21 for delection including complete blood count, electrolyte, APTT, PT, Fg, blood coagulation factors, and thromboelastography(TEG).

RESULTSThe levels of Hb, WBC, Plt, sodium and potassium in each sample accorded with standard of storing whole blood. The level of Hb, WBC, Plt and Na decreased along with prolonging of storage time, while the K level increased along with prolonging of stored time, APTT and PT prolonged along with prolonging of thored time, PT>17 min at d 21, the Fg level change was no-obvious, The level of factor Ⅴ and Ⅷ decreased more than 50 % of baseline on d 6 and 4 respectively; the levels of factor Ⅱ, Ⅶ, Ⅸ, Ⅹ, Ⅺ, Ⅻ showed decreasing trend, but their levels were less than 40 % of baseline values at d 21. TEG test showed that no abnormalily of R value was found, the abnormal valnes of K and Angle were observed at d 21, the abnormal value of MA was observed at d 14.

CONCLUSIONThe whole blood stored for 10 days possesses normal coagulation function showing important significance for treatment of hemorrhage from war injury and surgical openation of heart and chest.

0bjective: To establish a three-dimensional finite element model of mandible and study the transverse displacement of proximal segment after Bilateral Sagittal Split Ramus Osteotomy (BSSRO) with different retrogression amounts during mastication. Methods: DICOM data of a skull model were processed with MIMICS and ANSYS software, reconstructing the 3D model including the teeth and temporomandibular joint in order to simulate BSSRO and evaluate the transverse displacement of proximal segment with different retrogression amounts during mastication. Results: The mean of proximal segment width change were 2.955 mm and 3.490 mm, when retrogression amounts of distal segmentwere 3 mm and 8 mm, respectively.No significant difference between the two groups were found (P=0.131). Meanwhile the displacement color scale of the 3D finite element models showed that the apparent transverse displacement distribution of the proximal segment was measured around the gonial area, decreased from the exterior to the interior. Conclusions The mandibular angle width was significantly expanded right after BSSRO. The masticatory muscle system and single cortical fixation system played an important role in expanding the width of proximal segment. However there was no correlation between the widening effect and retrogression amounts of distal segment of mandible.

To explore the protective effect of naringin(Nar) on the injury of myocardium tissues induced by streptozotocin(STZ) in diabetic rats and the relationship with oxidative stress and endoplasmic reticulum stress(ERS)， the male SD rats were intraperitoneally injected with streptozotocin(STZ， 60 mg·kg⁻¹) to establish the diabetic rat model and then randomly divided into the type 1 diabetic rat group(T1DR)， the low-dose Nar group(Nar25)， the middle-dose Nar group(Nar50) and the high-dose Nar group(Nar100). The normal rats were designed as control group(Con). Nar25， Nar50， Nar100 groups were orally administered with Nar at the doses of 25.0， 50.0， 100.0 mg·kg⁻¹ per day， respectively， while the normal group and the T1DR group were orally administered with saline. At the 8th week after treatment， fasting plasma glucose and heart mass index were measured. The pathological changes in myocardial tissues were observed by microscope. The cardiac malondialdehyde(MDA) level and superoxide dismutase(SOD) activities were measured. The gene and protein expressions of glucose-regulated protein 78(GRP78)， C/EBP homologous protein(CHOP)， cysteinyl aspartate-specific proteinase 12(caspase 12) were detected by qRT-PCR and Western blot. According to the results， compared with control group， the myocardial structure was damaged， the content of MDA was increased， while the activities of SOD were decreased(<0.05) in T1DR group. GRP78， CHOP and caspase 12 mRNA and protein expressions were increased significantly in T1DR group(<0.05， <0.01). Compared with T1DR group， myocardial structure damage was alleviated in Nar treatment group. The content of MDA was decreased， while the activities of SOD were increased significantly. The mRNA and protein expressions of GRP78， CHOP and caspase 12 were increased， especially in middle and high-dose groups(<0.05， <0.01). After treatment with Nar for 8 weeks， myocardial structure damage was obviously alleviated in Nar treatment groups. The content of MDA was decreased， while the activities of SOD were increased significantly in myocardial tissues. The mRNA and protein expressions of GRP78， CHOP and caspase 12 were increased， especially in middle and high-dose groups(<0.05， <0.01). The findings suggest that Nar may protect myocardium in diabetic rats by reducing mitochondrial oxidative stress injuries and inhibiting the ERS-mediated cell apoptosis pathway.
Animals ; Apoptosis ; Cardiotonic Agents ; pharmacology ; Caspase 12 ; metabolism ; Diabetes Mellitus, Experimental ; Diabetic Cardiomyopathies ; drug therapy ; Endoplasmic Reticulum Stress ; drug effects ; Flavanones ; pharmacology ; Heat-Shock Proteins ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Transcription Factor CHOP ; metabolism

The objective of this study was to investigate Babesia infection among domestic animals in Western Yunnan Province and provide scientific evidence for developing control measures.A total of 1 073 domestic animals blood samples (274cattle,395 sheep,354 dogs,33 horses and 17 donkeys) were collected in 12 counties in Western Yunnan Province.Genomic DNA was extracted and a near full-length 18S rRNA gene sequence of Babesia was amplified by using nested PCR.Babesia species was identified by Blast program and phylogenetic tree.It was indicated that 50 samples were infected with Babesia,belonging to 5 species and with the infection rate of 4.66％.Among 274 cattle blood samples,11 were infected with Babesia (4.01％).Four of them were Babesia bovis and seven of them were Babesia bigemina.Among 395 sheep blood samples,38 were infected with Babesia (9.62 ％),37 of them were Babesia odocoilei-like parasites and 1 of them was Babesia capreoli-like parasites.Horses and donkeys were negative.In conclusion,domestic animals in Western Yunnan Province are infected with many kinds of Babesia,which threaten stock raising development and human health.It is necessary to strengthen prevention of babesiosis and investigate infection rate of babesiosis in human.

Objective: To evaluate the surgical prediction accuracy of Proplan CMF software for zygomatic reduction surgery using L-shaped osteotomy. Methods: Pre-and-postoperative 1-year CBCT data of 26 patients with zygomatic arch hypertrophy were imported in Proplan CMF software during 2014 Jan. to 2016 Jun., the 3D models were reconstructed for simulation of L-shaped osteotomy, characteristic landmarks were selected and 3D point measurement system was established. The measurement result were analyzed by one-way ANOVA. Meanwhile, the overlap color grading charts of preoperative and simulated images were also observed. Results: The facial width, bilateral zygomatic process angle and facial width index were [(128.56±2.72) mm, (106.87±2.53)°, (108.56±3.02)°and 1.41±0.03] in postoperative result, [(129.49±2.26) mm, (108.68±2.40)°, (108.85±3.02)°and 1.42±0.03]in simulated result and [(135.45±2.45) mm, (102.50±2.60)°, (103.41±2.56)°and 1.48±0.05] in preoperative result, with significant difference between preoperative and postoperative result, or between preoperative and simulated result (P<0.05), while no significance between postoperative and simulated result (P>0.05). The soft tissue zygomatic process distance was(153.25±2.58) mm in preoperative result, (150.23±2.76)mm in postoperative result , (149.36±3.27)mm in simulated result, with no significance between any of two groups result (P>0.05). The zygomatic process distance and bilateral zygomatic process tragal distance were (126.35±2.56) mm, (68.75±2.15) mm and(68.86±3.21) mm in postoperative result, showing significant differences compared with preoperative result [(120.16±3.18) mm, (74.58±3.19) mm and(76.14±3.15) mm] and simulated result [(118.86±3.45) mm, (73.85±3.57) mm and(76.87±2.58) mm] respectively(P<0.05), while zygomatic arch distance was not statistically different among the three groups(P>0.05). It indicated that predictive accuracy of facial width, facial width index, zygomatic process angle, soft tissue zygomatic arch distance was high but the soft tissue zygomatic process distance and zygomatic process tragal distance was relatively low. Meanwhile, the color overlay image showed that predictive accuracy was not good in the zygomatic region while the zygomatic arch area was high. Conclusions The predictive accuracy of Proplan CMF software for zygomatic arch hypertrophy is relatively high except for the zygomatic region. Further improvement of the CMF software is needed.