Objective To investigate the expression of Toll-like receptor 4 (TLR4) and CD80 in unexplained recurrent spontaneous abortion and their relationship,discuss its possible roles in guiding eugenics,and provide a new idea for clinical diagnosis and prevention.Methods The study group were 60 cases with unexplained recurrent spontaneous abortion patients.The control group included 30 cases of normal early pregnancy without previous history of adverse pregnancy.The expressions of TLR4 and CD80 were detected with immunohistochemistry in the peripheral blood,chorion,and decidua from each case.Results TLR4 had higher expression in the peripheral blood,the chorion and decidua from the recurrent spontaneous abortion women than that from normal pregnancy women (P ＜ 0.05).Levels of CD80 were significantly higher in recurrent spontaneous abortion group than in normal pregnancy group (P ＜ 0.05).There was a positive correlation between TLR4 and CD80 (r =0.982,0.986,and 0.776,P ＜0.01).Conclusions The study shows that TLR4 and CD80 were expressed in peripheral blood lymphocytes and villi and decidua of early pregnancy (normal pregnancy or abnormal pregnancy).Higher expressions of TLR4 and CD80 might play an important role in the occurrence of recurrent spontaneous abortion.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.

Objective　To study the relationship between gene's mutation in sputum cells and smoking consumption of patients with lung cancer. Methods　Sputum dispose liquid was added into 0.5?ml sputum, DNA was extracted twicely with saturated phenol-chloroform and ethanol precipitate; p53 and K-ras gene mutations were detected by using SSCP-PCR and RFLP-PCR methods. Results　Seventy-one of 110 patients with lung cancer were heavy smokers (index of smoking≥400), 55 of 71(77.5％) patients with heavy smoking were detected with gene alterations of p53 or K-ras (P＜0.05). Average indexes of smoking in the patients with p53 or K-ras were 861 and 630 respectively, whereas those in non-smokers were 284 and 554 (χ2＝36.56，P＝0.002). Conclusion　Detection of oncogene alterations in sputum is a simple method to practice. Oncogene alteration in smoking patients with lung cancer is higher, especially in heavy smokers, than those with no smoking. The results show that heavy smoking is one of the causes in oncogene mutations of bronchus and further investigation is necessary.

Objective: To explore the relationship between family cohesion, psychological resilience and non-suicidal self-injury among middle school students. Methods: By using the stratified random cluster sampling method, 2 065 students from 4 middle school from Fujian province were investigated with family cohesion scale, adolescent psychological resilience scale and adolescent self-injury behavior questionnaire. Results: The report rate of self-injury was 40.34%, which differed by childhood migration experiences, parenting style as well as academic pressure (χ2=11.66，29.45，12.48，P<0.01). Total scores and dimensional scales in family cohesion and psychological resilience showed significant differences in students with or without self-injury (t=-8.33，-12.08，-7.29，-11.53，-3.38，-7.37，-7.68，P<0.01). The family cohesion was positively correlated with the psychological resilience and each dimension (r=0.27-0.56, P<0.01), but negatively correlated with non-suicidal self-injury (r=-0.18, P<0.01). The psychological resilience and each dimension were negatively correlated with non-suicidal self-injury (r=-0.24--0.14, P<0.01). Psychological resilience had a partial mediating effect between family cohesion and non-suicidal self-injury, with mediating effect accounting for 61.11% of the total effect. Conclusion Psychological resilience played a mediating role between family cohesion and non-suicidal self-injury. By promoting family cohesion to improve psychological resilience, it is helpful to prevent non-suicidal self-injury among middle school students.

BACKGROUNDTo investigate the clinical possibility of detection of lung cancer cells in the peripheral blood with cytokeratin and 2F7/S5A monoclonal antibody by flow cytometry (FCM) in patients with lung cancer.

METHODSFive mililiter peripheral blood was collected. The mononuclear cells were isolated by Ficoll-Hypaquegradient centrifugation. Then the cells were labeled with lung specific antibody (2F7/S5A) and antibodies against CD45, cytokeratin (CK). The quantitation of CD45 -CK +2F7/S5A + cells were performed by FCM.

RESULTSA total of 151 people were studied, including 20 healthy controls, 116 patients with lung cancer and 15 patients with benign pulmonary diseases. The phenotype of monocytes of peripheral blood was CD45 +CK -2F7/S5A -, while the phenotype of lung cancer cells was CD45 -CK +2F7/S5A +. No cancer cell was found in peripheral blood of all the healthy controls. One out of 15 cases of benign pulmonary diseases was found to have CD45 +CK -2F7/S5A - cells in peripheral blood. And 39 out of 116 patients with lung cancer were found to have circulating lung cancer cells. The sensitivity, specificity, positive predictive value and negative predictive value of detection was 33.62% , 93.33%, 97.50% and 15.38%, respectively. The average quantitation of circulating cancer cell detected was 0.12×10⁶/L.

CONCLUSIONSThere is high specificity and positive predictive value of the examination of circulating cancer cells in patients with lung cancer by FCM, which could detect trace cancer cell in peripheral blood of lung cancer. It may be valuable in diagnosis and treatment of lung cancer.

Objective: To investigate the characteristics and prognostic factor of central nervous system (CNS) involvement in patients with hemophagocytic lymphohistiocytosis (HLH) . Methods: From January 2006 to October 2015, 152 patients with HLH, 88 patients had CNS involvement, their clinical data were collected, and survival was analyzed using the Kaplan-Meier life table method, univariate and multivariate Cox regression model analyses were applied to identify the risk factors of prognosis. Results: ①57.9% patients complicated with neurological symptoms, cerebrospinal fluid abnormalities were observed in 37.0% patients, 57.5% patients had abnormal neuroradiology. ②36 patients survived well, 3 patients lost to follow-up, 49 dead, 1 survival patient had epilepsy. ③The 3-year overall survival rate of 88 patients was 44%. ④abnormal CSF and unreceived IT bore a significant, independent adverse prognostic value (P<0.05) . Conclusion CNS involvement in HLH has a high frequency and poor prognosis, few patients remained neurologic sequelae; abnormal CSF related to poor prognosis, positive intrathecal injections could improve the prognosis.

Objective To investigate the clinical features and prognosis of central nervous system(CNS)in-volvement in Epstein-Barr virus(EBV)associated hemophagocytic lymphohistiocytosis(HLH).Methods A total of 89 patients with EBV-HLH diagnosed in Children's Hospital of Chongqing Medical University from June 2006 to Octo-ber 2015 were divided into involved CNS group and non-involved group according to whether there was CNS involve-ment. The clinical manifestations,laboratory examinations and outcomes of these two groups were analyzed. Results Among these 89 patients with EBV-HLH,39 patients developed CNS disease,19 cases of them had neuro-logical symptoms or signs,including convulsions,unconsciousness,facial palsy,dysarthria,dysphagia,irritability,neck stiffness,Babinski sign positive,opisthotonus;9 cases of them had abnormal cerebrospinal fluid(CSF),with elevated white blood cell count and protein level;26 patients had abnormal brain images,including deepen or widening cortical sulci,atrophy,hemorrhage,high T2 signal in magnetic resonance imaging(MRI).The 3-year survival rate in involved CNS group was lower than those of non-involved group(66.7% vs.86.0%),and there was statistical significance (χ2=4.267,P=0.039).The involved CNS group had higher ferritin(χ2=5.092,3.921;P=0.024,0.048)and lower platelets(Z= -2.643,P=0.008)than those of non-involved group,and there were statistical significances.COX mul-tivariate analysis showed that neurological symptom and abnormal CSF were independent prognostic factors(RR=3.134, 3.339,all P<0.05).Conclusion CNS involvement is frequent in EBV-HLH.The prognosis of children with involved CNS group is worse than those of non-involved group.Neurological symptoms and abnormal CSF are related to poor prognosis.

OBJECTIVETo explore the clinical phenotype of a family affected with congenital dysfibrinogenemia and potential mutations underlying the disease.

METHODSCoagulation testing and hepatorenal function testing were conducted on 18 individuals from three generations. Plasma fibrinogen was extracted and analyzed with SDS-PAGE electrophoresis. All of the exons and flanking sequences of fibrinogen FGA, FGB, FGG genes were analyzed by PCR, and the products were subjected to Sanger sequencing.

RESULTSHepatorenal function, prothrombin time and activated partial thromboplastin time of the proband were all normal. However, his thrombin time was significantly prolonged. Fibrinogen activity was decreased, while the concentration of antigen was in the normal range. The results of his mother, brother, and nephew were similar. DNA sequencing has confirmed that the proband, his mother, brother, and nephew have all carried a g.5877G>A mutation in the exon 8 of the FGG gene, which resulted in replacement of arginine (Arg) by histidine (His) at position 275.

CONCLUSIONThe Arg275His mutation of the fibrinogen gamma chain probably underlies the pathogenesis of congenital dysfibrinogenemia in this family.

Adult ; Afibrinogenemia ; genetics ; metabolism ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Female ; Fibrinogen ; genetics ; metabolism ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; Pedigree ; Point Mutation

Objective:To validate the morphological changes of the parieto-occipital sulcus on the transcalvarial axial plane between 20 and 32 weeks of gestation, simplify grade for assessing fetal parieto-occipital sulcus development, and confirm its clinical feasibility.Methods:This was a cross-sectional study analysis that included 550 cases of normal singleton fetuses between 20 and 32 weeks of gestation, who underwent routine ultrasound examinations at Shenzhen Maternity and Child Healthcare Hospital from September 2019 to June 2022. The morphological changes of the bilateral parieto-occipital sulci on the transcalvarial axial plane were observed. The development of the parieto-occipital sulcus was classified into 6 grades based on the developmental features of angulation, progressive closure, and curvilinear growth: straight or shallow arcuate (Grade 0), shallow and wide V-shaped (Grade 1), deep and narrow V-shaped (Grade 2), Y-shaped (Grade 3), I-shaped (Grade 4), and curvilinear (Grade 5). The gestational age at examination and pregnancy outcomes were recorded. The distribution of gestational weeks for fetuses with different grades of parieto-occipital sulci on the left and right sides was analyzed. The symmetry between bilateral parieto-occipital sulcus gradings within individuals, as well as the inter-observer and intra-observer reliability were assessed using the Weighted Kappa coefficient. The gender differences in asymmetry of parieto-occipital sulci grades between the left and right sides was analyzed. Moreover, a model for predicting the grade of the parieto-occipital sulcus based on gestational week was established.Results:Grade for the left parieto-occipital sulcus was obtained for 549 fetuses, while grade for the right was obtained for 550 fetuses. From 20 to 32 weeks of gestation, the morphology of the fetal parieto-occipital sulcus was divided into Grade 0-5, progressing from low to high with gestational development. Grade 0 showed that the sulcus was not visible or only had a slight arcuate indentation, occurring at 20-22 weeks; Grade 1 presented as a shallow and wide "V" shape with an obtuse angle at the top, appearing from 20 to 27 weeks; Grade 2 was a deep and narrow "V" shape with an acute angle at the top, appearing from 24 to 29 weeks; Grade 3 appeared as a "Y" shape with the top part partially closed and the bottom still open, occurring between 26 to 30 weeks; Grade 4 was a fully closed "I" shape, appearing at 29-32 weeks; Grade 5 presented as a curved shape, indicating the parieto-occipital sulcus was approaching maturity, appearing from 31 to 32 weeks. There was no statistically significant difference in the distribution of gestational weeks for bilateral parieto-occipital sulcus developmental grade ( P>0.05). Bilateral parieto-occipital sulcus grade could be assessed in 549 fetuses, of which 43 cases (7.83%) exhibited grade asymmetry with a one-grade difference between sides; such asymmetry showed no significant difference between male and female genders ( P=0.647). The weighted kappa coefficient analysis results indicated a strong consistency in the development of the parieto-occipital sulci on both sides within individuals, generally demonstrating symmetrical development ( P<0.001). The intra-observer and inter-observer weighted kappa coefficients were 0.92 and 0.75, respectively, with good consistency. Conclusions:Prenatal ultrasound via the transcalvarial axial plane enables a preliminary and rapid assessment of the development of bilateral parieto-occipital sulci, facilitating early evaluation of fetal cortical maturation.

Objective:To study the stability and morphological changes of the convexity sulci in normal fetuses between 20 and 32 weeks, and to explore the simplified grade for evaluating the convexity sulci development and analyzing its clinical significance.Methods:This study was a cross-sectional analysis. A total of 551 cases of normal singleton pregnancies between 20 and 32 weeks of gestation were retrospectively collected, who underwent routine ultrasound examinations at Shenzhen Maternity and Child Healthcare Hospital from September 2019 to June 2022. The display of the far-field convexity sulci on the transcalvarial axial plane was observed as 0 for not displayed and 1 for displayed.Further, based on the morphology and number of convexity sulci, they were classified into five grades: no sulcus displayed (grade 0), one sulcus (grade 1), two sulci (grade 2), three sulci (grade 3), and four or more sulci (grade 4). The gestational age at examination and pregnancy outcomes were recorded. The distribution characteristics of gestational weeks for each grade of the convexity sulci were analyzed, and the gestational week distribution of the left and right convexity sulci was compared to analyze bilateral symmetry. The Weighted Kappa coefficient was used to analyze inter-observer and intra-observer consistency, and curve regression analysis was employed to establish a model for predicting grade based on gestational weeks.Results:Before 25 weeks of gestation, the convexity of the fetal cranial vertex was completely smooth.The central sulcus consistently appeared after 26 weeks, while the superior frontal sulcus, intraparietal sulcus, postcentral sulcus, and precentral sulcus consistently appeared between 28 and 31 weeks. Among these, the superior frontal sulcus had a lower display rate before 29 weeks. By 32 weeks, all convexity sulci of the cranial vertex should be visible. Three hundred and eleven fetuses were graded for the left, and 240 fetuses were graded for the right. The developmental grade of the convexity sulci increased from Grade 0 to Grade 4 as the gestational age progressed. Grade 0 appeared between 20-26 weeks, grade 1 between 25-28 weeks, grade 2 between 26-28 weeks, grade 3 between 27-30 weeks, and grade 4 between 27-32 weeks. The distribution of grade did not differ significantly between the left and right sides of grade 0, 1, 3 and 4 (all P>0.05), while there was a significant difference in the distribution of gestational age between the convexity sulci of grade 2 ( P<0.05). The Weighted Kappa coefficients for intra-observer and inter-observer consistency were 0.94 and 0.86, respectively, indicating strong consistency. Conclusions:The simplified grade for assessing the development of convexity sulci in normal fetuses on the transcalvarial axial plane via prenatal ultrasound can provide a preliminary evaluation of the maturation of convexity sulci in fetuses between 20 and 32 weeks of gestation.