Objective To evaluate the application value of bedside lung ultrasound in diagnosis of neonatal severe pneumonia.Methods Bedside lung ultrasound and chest X-ray were performed in 59 the newborn patients admitted to the newborn department for suspected severe pneumonia,from October 2015 to March 2016.According to the characteristics of the chest X-ray,patients were divided into consolidation group (widely spot shadow in double lung and airbronchograms) and the unconsolidation group (enlargement,fuzzy texture in double lung or small patch sample changes) Differences in ultrasonic characteristics were compared between the two groups.Results Ultrasonic image characteristics were pulmonary consolidation with air bronchograms,A-line disappearance,pleural line abnormalities.Among 23 patients with chest X-ray,21 patients were found pulmonary consolidation by lung ultrasound,and all 36 patients in unconsolidation group were not found pulmonary consolidation by lung ultrasound,and there was no significant difference in the detection rate by lung ultrasound between the two groups (P＞0.05) The pulmonary ultrasound results of 20 cases in the consolidation group showed A-line disappearance,37 cases in the unconsolidation group showed A-line disappearance.There was no statistical significant differences between the two groups (P＞0.05).The pulmonary ultrasound results showed 11 cases in the consolidation group and 15 cases in the unconsolidation group with pleural line abnormalities,there was no statistically significant differences between the two groups (P＞O.05).Conclusions Ultrasound image characteristics of neonatal severe pneumonia include pulmonary consolidation and air bronchograms,A-line disappearance,pleural line abnormalities.Compared lung ultrasound and chest X-ray result,there was no difference between consolidation group and unconsolidation group.Both of the two methods can be used for evaluating the diagnosis of pneumonia.

Objective To analyze the clinical characteristic of Chinese autoimmune pancreatitis (AIP) patients.Methods All clinical data of 81 patients with a diagnosis of AIP in Shanghai Changhai Hospital from February 2005 to May 2012 were analyzed.Results The sex ratio was 7.1∶1 and the mean age was (57± 12) years old in 81 patients with AIP.Obstructive jaundice was the initial symptom in 51.9％ (42/81) patients.In patient receiving CT,focal and diffuse type accounted for 45 and 35 patients.respectively,and pseudocyst was the main manifestation in 1 patient,biliary tract was involved in 59(72.8％ ) patients,dilatation of main pancreatic duct was observed in 5 ( 11.1％ ) patients.In patients receiving PET-CT,diffuse increased Flourine-18 FDG uptake by the pancreas was found in 11 patients,focal increased uptake in 2patients,and significant extra-pancreatic uptake was found in 5 patients.The positive rate of serum IgG4,CA19-9,ss DNA,anti-nuclear antibody and ds-DNA antibody was 94.6％ (53/81),54.4％ (37/68),14.3％ (4/28),10.7％ (3/28),7.1％ (2/28),respectively.The pathological findings of H-E staining and IgG4 immunohistochemical analysis in 20 patients were consistent with lymphoplasmacytic sclerosing pancreatitis.Conclusions Type 1 AIP is the main subtype of AIP in China.Combining clinical symptoms,extra-pancreatic manifestations,imaging or nuclear medicine findings,serology,cytology or histology can effectively increase the correct diagnosis rate of AIP.

Objective: To evaluate the safety of enterovirus 71（EV71）inactivated vaccines produced by Wuhan Institute of Bio Products through learning the incidence of adverse events following immunization（AEFI）in children aged 6 months to 3 years old after vaccination. Methods: According to the national requirement for vaccine safety monitoring program，data of AEFI cases after inoculated EV71 vaccines from September 2017 to October 2018 in Zhejiang Province was collected by combining active and passive monitoring. Demographic characteristics，space distribution，dose and incidence of AEFI were analyzed. Results: A total of 107 503 children were included in this study，of which 27 173 were actively monitored and 80 330 were passively monitored. The monitoring results showed that 288 cases of AEFI occurred after inoculated EV71 vaccines，the reported incidence rate was 267.90/100 000. Of 288 cases，266 cases had common reactions（247.43/100 000），14 cases had abnormal reactions（13.02/100 000）and 8 cases had complications（7.44/100 000）；145 cases were passively monitored（180.51/100 000）and 143 cases were actively monitored（526.26/100 000）；150 cases were males and 138 cases were females，with the males to females ratio of 1.09∶1. The AEFI of EV71 vaccines were reported in all the cities of Zhejiang Province，with Jinhua，Taizhou and Hangzhou ranking the top three and accounting for 50.69% of all the reported cases. About 202 cases of AEFI（70.14%）occurred after the first dose of EV71 vaccines，and 163 cases（56.60%）occurred on the day of inoculation.Most of common reactions were fever，swelling and sclerosis. The harm of the reported abnormal reactions was mild，with 8 cases of anaphylactic rash and no other serious abnormal reactions were found. Conclusion The reported incidence rate of AEFI of the domestic EV71 vaccine in children aged from 6 months to 3 years is 267.90/100 000，without serious abnormal reactions，suggesting that the EV71 vaccine is safe.

Objective : To investigate the awareness of human papillomavirus ( HPV ) vaccine and willingness to vaccinate daughters among parents of primary and middle school students, so as to provide the reference for the promotion of HPV vaccine in primary and middle school girls. Methods : Using multi-stage stratified cluster sampling method, the parents of girls in in Grade Four to Nine from schools in Gongshu District of Hangzhou, Xiuzhou District of Jiaxing and Wuxing District of Huzhou were selected. A questionnaire survey was conducted to collect demographic information, HPV vaccine related knowledge and willingness to vaccinate daughters with HPV vaccines. The multivariate logistic regression model was used to analyze the influencing factors for the willingness to vaccinate daughters with HPV vaccines among parents. Results : Totally 1 500 questionnaires were sent out, and 1 466 were effectively collected, with an effective rate of 97.73%. There were 313 fathers responded, accounting for 21.35%; and 1 153 mothers responded, accounting for 78.65%. The awareness rate of HPV vaccine was 16.81%. The rate of willing to vaccinate daughters with HPV vaccines was 49.86%. The multivariate logistic regression analysis showed that the patients who ever vaccinated daughters with self-paid vaccines ( OR=1.935, 95%CI: 1.473-2.541 ), knew cervical cancer ( OR=1.424, 95%CI: 1.065-1.904 ), knew HPV vaccine dose ( OR=1.672, 95%CI:1.216-2.301 ), knew the best vaccination period ( OR=1.392, 95%CI: 1.032-1.876 ), knew the need of cervical cancer screening even after vaccination ( OR=1.596, 95%CI:1.227-2.075) were more willing to vaccinate daughters with HPV vaccines, while the parents who thought HPV vaccine expensive ( OR=0.154, 95%CI: 0.099-0.240 ) were less willing to vaccinate daughters with HPV vaccines. Conclusions The rates of HPV vaccine awareness and willingness to vaccinate daughters are 16.81% and 49.86% among parents of primary and middle school students. Their knowledge of HPV vaccine and the price of the vaccine may affect their willingness to vaccinate daughters.

OBJECTIVETo analyze the deletion region for two fetal cases with large Yq deletions in order to provide genetic counseling and prenatal diagnosis.

METHODSFor both cases, amniotic fluid samples were cultured and analyzed with G banding and fluorescence in situ hybridization (FISH). Multiplex polymerase chain reaction was also carried out to amplify 15 sequence tagged sites (STS) of azoospermia factor (AZF) on the Y chromosome.

RESULTSFor both samples, the karyotypes were determined as 46,X,del(Y)(pter→q11:). No heterochromatin was found in C band. The karyotypes of their fathers were 46,XY, and heterochromatin was found in C band. STS analyses suggested that only sY82, sY84 and sY86 in AZFa were amplifiable while the other 12 STS were negative in amniotic fluid for the first case, which indicated deletions of AZFb, AZFd and AZFc. No AZF deletion was found in its father. For the second case, all 15 STS were amplifiable in the amniotic fluid, suggesting no AZF deletion. No AZF deletion was found in its father too.

CONCLUSIONConventional karyotyping combined with FISH and molecular genetics techniques can enable characterization of AZF microdeletions and facilitate genetic counseling and prenatal diagnosis.

Adult ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Counseling ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo explore the origin and mechanism of small supernumerary marker chromosomes (sSMC) in order to facilitate genetic counseling.

METHODSChromosome karyotypes of two fetuses and their immediate family members were analyzed by conventional G banding. High-throughput whole genome sequencing was used to determine the origin of sSMCs.

RESULTSFetus 1 was shown to have a karyotype of 47,XY,+mar but with normal FISH and B ultrasound findings. Its father also had a 47,XY,+mar karyotype with normal FISH results and clinical phenotype. High-throughput genome sequencing revealed that fetus 1 and its father were both 46,XY,dup(21)(q11.2;q21.1) with a 6.2 Mb duplication of the long arm of chromosome 21. The fetus was born with normal phenotype and developed well. Its grandmother also had a karyotype of 46,XX,t(15;21)(q13;p13) with normal FISH result and clinical phenotype. The karyotypes of its mother and grandfather were both normal. Analysis of fetus 2 showed a 47,XY,+mar karyotype with normal FISH results. High-throughput genome sequencing suggested a molecular karyotype of 46,XX. The fetus was born with normal phenotype and developed well. The karyotypes of its parents were both normal.

CONCLUSIONConsidering their variable origins, identification of sSMC should combine conventional G banding analyses with high-throughput whole genome sequencing for precise delineation of the chromosomes.

Adult ; Amniotic Fluid ; chemistry ; Chromosome Banding ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; Cytogenetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Markers ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis ; Young Adult

OBJECTIVETo track and analyze two false positive cases from non-invasive prenatal testing for potential fetal aneuploidy.

METHODSThe two cases, respectively reported to have XO (+++) and T18 (1/20) XO(+), were analyzed with conventional karyotyping, fluorescence in situ hybridization (FISH) and massively parallel genomic sequencing (MPS).

RESULTSThe first fetus, who was suspected for XO(+++), was verified to have super female syndrome (47,XXX/46,XX) due to confined placental mosaicism by karyotyping of amniotic fluid cells, FISH analysis of placenta and massively parallel sequencing (MPS) of fetal tissue. The second fetus, suspected to have trisomy 18 (1/20) XO(+), was verified to have Turner syndrome by karyotyping, FISH and MPS analyses of umbilical cord blood cells. And the karyotype was 45,X[48]/46, X, der(X) del(X) (p11.21) del(X) (q13.3)[62].

CONCLUSIONNon-invasive prenatal testing carries a risk for false positive diagnosis of fetal sex chromosome and trisomy 18. Combined cytogenetic and molecular techniques are required to ensure an accurate diagnosis.

Adult ; Aneuploidy ; Chromosome Aberrations ; Diagnostic Errors ; False Positive Reactions ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Pregnancy ; Prenatal Diagnosis ; Young Adult

OBJECTIVE: To delineate cytogenetic and molecular abnormalities of a fetus carrying a de novo 46,X,der(X),t(X;Y)(p22.3;p11.2). METHODS: G-banded karyotyping and next-generation sequencing (NGS) were used to analyze the fetus, his father and sister. Single nucleotide polymorphism-based arrays (SNP-array), multiple PCR and fluorescence in situ hybridization (FISH) were utilized to verify the result. RESULTS: G-banded karyotyping at 320 bands showed that the fetus had a normal karyotype, while NGS has identified a 3.58 Mb microdeletion at Xp22.33 and a Y chromosomal segment of about 10 Mb at Yp11.32p11.2. With the sequencing results, high-resolution karyotyping at 550-750 bands level has determined the fetus to be 46,X,der(X)t(X;Y)(p22.3;p11.2). The result was confirmed by PCR amplification of the SRY gene, FISH and SNP-array assays. The karyotypes of his father and sister were both normal. His sister also showed no amplification of the SRY gene, and her NGS results were normal too, suggesting that the karyotype of the fetus was de novo. CONCLUSION Combined karyotyping, NGS, SNP-array, PCR and FISH assay can facilitate diagnosis of XX disorder of sex development.
Chromosomes, Human, X ; genetics ; Disorders of Sex Development ; genetics ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Translocation, Genetic

Objective: To evaluate the post-marketing safety of inactivated Enterovirus type 71 (EV-A71) vaccine (human diploid cell) . Methods: A total of 20 191 healthy children aged 6 to 59 months were invited to receive 2 doses of EV-A71 vaccine in Zhejiang Province from September 2016 to December 2017. Child caregivers were followed up on the 4^th or 5^th day after each EV-A71 vaccination, and the incidence of local, systemic, and other adverse events within 3 days after vaccination was recorded to assess vaccine safety. Describe the differences in adverse events among children with different characteristics. Results: A total of 32 230 doses were observed in this study, of which 20 191 and 12 039 were vaccinated for the first and the second dose, respectively; and the incidence of adverse events within 3 days was 2.045% (413 doses) and 1.611% (194 doses), respectively. After the first and the second dose, the number of systemic adverse events was the highest, 371 and 175 cases, respectively, with an incidence of 1.837% and 1.454%, respectively; the number of local adverse events was the lowest, 14 and 2 doses, respectively, with an incidence of 0.069% and 0.017%. Local adverse events occurred after vaccination were generally mild, and only 2 patients had level of 3; among the systemic adverse events, 39 patients had a fever level of 3 or higher, accounting for 8.2% of the total fever. Most of the symptoms in the local adverse events did not require treatment, only 3 cases of vaccination site rash and 2 cases of pruritus were self-purchased drugs or outpatient treatment; except for 5 cases of fever, the other symptoms were not hospitalized in the case of systemic adverse events. Conclusion The incidence of adverse events within 3 days after vaccination with EV-A71 vaccine was low in children, mainly systemic adverse events, and the prognosis was good.

Bombyx mori is a lepidopteran insect with important economic value. Bombyx mori nucleopolyhedrovirus (BmNPV) causes huge economic loss to silkworm industry in China every year. The objective of this study is to determine the anti-BmNPV mechanism of Bombyx mori strain NC99R, and to provide a basis for understanding the molecular mechanism of the silkworm resistance strain. The normal control Dazao (DZ) strain and the NC99R resistant strain were fed with occlusion bodies (OB). The median lethal dose (LD50) analysis of the DZ and NC99R showed that the LD50 of DZ was 1.2×10⁵ OBs/larva, while NC99R was 1.8×10⁶ OBs/larva. The LD50 of the NC99R was about 15 times higher than the DZ. The mortality of DZ and NC99R were analyzed, which were fed with 1×10⁶ OBs/larva and injection with 1×10⁶ BVs/larva. The results showed that the death peak of DZ was concentrated in the 4th to 6th day. And the death peak of NC99R was concentrated in the 6th to 8th day, with a delay of 1-2 days compared with the control. The BmNPV DNA copy number showed that the BmNPV genome in DZ proliferated rapidly. The copy number of BmNPV DNA in NC99R were increased slowly after oral infection and body injection. HE staining showed that midgut tissue has no significant difference between DZ and NC99R in the early stage of oral infection. At 96 h p.i., the nucleus of DZ midgut became larger and shedding. The NC99R had enlarged nuclei, but the cells were still arranged neatly. Finally, the expression of virus genes in different periods were analyzed by RT-PCR. The results indicated that the immediate early gene ie-1 expression levels began to down-regulate after 24 h p.i.. The early, late, and extremely late genes were also down-regulated, and finally maintained at a lower expression level.