Objective:To observe the inhibitory effects of Huangqi Jiedu Decoction on lung metastasis of breast cancer in nude mice; To explore the mechanism of intervening epithelial mesenchymal transformation (EMT) induced by Wnt/β-catenin signaling pathway.Methods:Totally 30 nude mice were divided into model group, adriamycin group and Huangqi Jiedu Decoction low-, medium-, and high-dosage groups according to random number table method. Each group was injected subcutaneously with mouse breast cancer 4T1 cells to construct tumor - bearing nude mice model. Huangqi Jiedu Decoction low-, medium- and high-dosage groups were intragastrically administrated with Huangqi Jiedu Decoction 17.82, 35.64 and 71.28 g/kg; adriamycin group was injected intraperitoneally adriamycin 0.05 g/kg; model group was intragastrically administrated with normal saline of the same volume for 21 d. Tumor volume was measured at 9, 15, and 21 days after modeling. After the end of administration, the tumor tissue was separated, the tumor weight was measured, and the tumor inhibition rate was calculated. The lung tissue was Isolated,, the number of lung metastatic nodules and the inhibition rate of lung metastasis was counted. HE staining was used to observe the tissue morphology and evaluate the effectiveness of the model. The protein expressions of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by Western Blot. The mRNA levels of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by real-time fluorescent quantitative PCR.Results:Compared with the model group, the tumor volume and mass of Huangqi Jiedu Decoction low-, medium- and high-dosage groups decreased ( P<0.01); the number of pulmonary metastasis nodules in Huangqi Jiedu Decoction high-dosage group significantly decreased ( P<0.01); the mRNA and protein expressions of β-catenin and Vimentinm decreased in the Huangqi Jiedu Decoction low-, medium- and high-dosage groups ( P<0.01), and the protein and mRNA expressions of E-Cadherin increased in the Huangqi Jiedu Decoction high-dosage group ( P<0.01). Conclusion:Huangqi Jiedu Decoction can effectively inhibit the growth and lung metastasis of breast cancer transplanted tumor, and the mechanism may be to down-regulate the expression of key molecules in the Wnt/β-catanin signaling pathway, thereby inhibiting the EMT process, so as to inhibit the lung metastasis of breast cancer.

OBJECTIVE: To present on a prenatally diagnosed case with complex structural rearrangements of chromosome 8. METHODS: Chromosome karyotyping, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) were carried out for a fetus with increased nuchal thickness. RESULTS: The karyotype of the amniotic fluid sample showed extra materials on 8p. FISH revealed a centromeric signal at the terminal of 8p with absence of telomeric signal. CMA revealed partial deletion of 8p23.3 [(208049_2256732)×1], partial duplication of 8p23.3p23.2 [(2259519_3016818)×3], and partial duplication of 8q [8q11.1q12.2(45951900_60989083)×3]. CONCLUSION The complex structural rearrangements of chromosome 8 in this case has differed from the commonly seen inv dup del(8p).
Female ; Pregnancy ; Humans ; Chromosomes, Human, Pair 8/genetics* ; In Situ Hybridization, Fluorescence ; Gene Rearrangement ; Prenatal Diagnosis ; Centromere

OBJECTIVE: To report on a case of mosaicism 13q inversion duplication, analyze its mechanism, and discuss the correlation between its genotype and phenotype. METHODS: Amniotic fluid and umbilical cord blood were collected at 23 and 32 weeks of gestation, respectively. Combined with G-banding chromosome karyotyping analysis, single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) were used to confirm the result. RESULTS: The karyotype of the fetus was determined as 47,XY,+inv dup(13)(q14.3q34)/46,XY. After careful counseling, the couple decided to continue with the pregnancy, and had given birth to a boy at 40 weeks' gestation. Except for a red plaque (hemangioma) on the nose bridge, no obvious abnormality (intelligence to be evaluated) was discovered. CONCLUSION To provide reference for clinical genetic counseling and risk assessment, the location and proportion of new centromere formation should be fully considered in the case of mosaicism 13q inversion duplication.
Amniocentesis ; Chromosome Inversion/genetics* ; Comparative Genomic Hybridization ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Mosaicism ; Pregnancy ; Prenatal Diagnosis

ObjectiveTo determine the epidemiological characteristics and risk factors of a norovirus GII.17 outbreak in Minhang District of Shanghai in 2018， and provide evidence for prevention and control measures of norovirus infection. MethodsDescriptive epidemiological analysis was performed. In addition， a retrospective cohort study was conducted to determine the risk factors. ResultsFrom May 30^th to June 1^st of 2018， a total of 132 cases （126 clinical cases and 6 confirmed cases） were documented，with an attack rate of 29.20%（132/452）.All cases were children in a kindergarten， with the average age of 5 years and 43.9% being male. The cases were reported in all the classes， with no clustering by class or floor．The epidemic curve was characterized by a point source exposure， which was estimated to be probably between 7 AM on May 30^thand 0：30 AM on May 31^st. The retrospective cohort study showed that the attack rate significantly differed between the children who had taken and did not take the school lunch （RR=∞） on May 30^th，and those who had taken and did not take seafood noodles （RR=4.11， 95%CI：1.09-15.55） （P<0.05）. Among a total of 73 specimens， six specimens collected in child cases and one specimen in an asymptomatic chef tested positive for GII.17 type of norovirus. In addition， one retained food specimen of seafood noodles was positive for Aeromonas hydrophila. Viral shedding in the asymptomatic chef remained over 30 days. ConclusionThe outbreak was caused by seafood noodles contaminated by norovirus. It warrants enhancement in the regulation of food safety in canteens and regular examination of norovirus infection in catering industry employees.

Objective:To develop a self-made plasma quality control material for non-invasive prenatal testing (NIPT) and evaluate its performance.Methods:139 NIPT-negative maternal plasmas stored in the genetic department of Shaoxing maternal and child health hospital from January 1, 2019 to June 30, 2021 were divided into male groups (19 cases) and female groups (120 cases) according to the neonatal gender. 9360 cases from September 2020 to September 2021 were enrolled as clinical validation cases.First step, 200 μl plasma from a 47 years-old non-pregnant healthy women was used as a matrix. Different amounts (0.1, 0.2, 0.5, 2.5, and 5 μl) of positive DNA from fetal chromosome aneuploidy (T21, T18, T13) detection kit were added. The appropriate volume of positive DNA was 0.5 μl according to the test results. Second step,Plasma in male and female group was treated as matrix. 0.5 μl positive DNA was added per 205 μl. Plasma matrix from female group showed good repeatability and the sensitivity was 100%.Third step, evaluate the self-made plasma quality control material, including storage stability, matrix uniformity and repeatability, and the effect of different batch numbers of positive DNA, by calculating Z score and the CV of fetal DNA concentration (FF).Results:Plasma matrix from female group showed good repeatability and the sensitivity was 100%, while the sensitivity of male group was only 84%. The CV of FF in female matrix was 3.9% in the repetitive experiments. After adding 0.5 μl positive DNA, the mean FF of self-made positive plasma quality control was 5.63%±0.42%, Z values>6, and the CV was 7% after storage of three months. Considering the concentration variation of positive DNA in different lots, 1 μl of positive DNA should be added when the FF of positive DNA is lower than 10%.Used in 9360 clinical cases from September 2020 to September 2021, all positive plasma quality control materials showed positive results, and the positive predictive value of trisomy 21 was 100%.Conclusions:The NIPT self-made positive plasma quality control material has been successfully developed in this study. The preliminary experimental results show that it has good repeatability and stability, which is suitable for clinical application.

OBJECTIVE: To detect common pathogenic variants associated with congenital deafness among neonates from Huizhou and surrounding areas and discuss its implications. METHODS: Thirteen hot-spot mutations in four most common pathogenic genes were screened among 20 934 neonates from March 2017 to December 2019. RESULTS: In total 760 neonates were found to carry common pathogenic variants (3.63%). Sixty two neonates have carried homozygous/compound heterozygous variants or homoplasmy/heteroplasmy mutations of mtDNA (0.29%). Further analysis of five abnormal cases revealed that 3 of them have carried compound heterozygous mutations of GJB2 gene, and 2 were due to compound heterozygous variants of the CDH23 gene. CONCLUSION Genetic testing has a great clinical significance for the prevention and reduction of congenital hearing loss, but the scope needs to be updated and redefined by removing mutation sites with a very low rate, adding new significant sites, and improvement of the technical strategies.
Connexin 26 ; Connexins/genetics* ; DNA Mutational Analysis ; Deafness/genetics* ; Genetic Testing ; Hearing Loss/genetics* ; Humans ; Infant, Newborn ; Mutation ; Neonatal Screening

Objective:To evaluate the immunogenicity and immune persistence of purified Vero cells rabies vaccine (PVRV) with Zagreb and Essen regimen.Methods:Prospective study: Patients with first Class II exposure to rabies were recruited from the Dog Injury Cinic of Guangxi Center for Disease Control and Prevention (Guangxi CDC) and randomly divided into the Zagreb (2-1-1) and Essen (1-1-1-1-1) regimen group. All patients were inoculated with the vaccines from the same manufacturer and batch, and 3 ml serum was collected at the 45th day and in 1-year, 2-year and 3-year after immunization. Rapid fluorescent inhibition test (RFFIT) was used to detect rabies virus neutralizing antibody (RVNA). The attenuation of RVNA positive rate and geometric mean titer (GMT) with time was analyzed. Retrospective study: The informed consents for rabies vaccine in the Dog Injury Clinic of Guangxi CDC were checked out. The patients who were injected with PVRV (same manufacturer but unlimited batch) but without passive immune agents for the first time within 3 years were selected and divided into 1-year, 2-year and 3-year group. Each group was further divided into Zagreb and Essen regimen group. The serum (3 ml) was collected at 1 year, 2 years and 3 years after immunization and detected the RVNA by RFFIT.Results:Prospective study: The RVNA positive rates on the 45th day and in 1-year, 2-year and 3-year after immunizationin in the Zagreb and Essen regimen group were 100%, 95%, 85%, 80% and 98.25%, 89.47%, 89.47%, 85.96%, respectively. There was no statistically significant difference in the RVNA positive rates at the same time point between the two regimen groups ( P>0.05). The RVNA GMT on the 45th day and in 1-year, 2-year, and 3-year in the Zagreb and Essen regimen group were 11.32 IU/ml, 1.69 IU/ml, 1.30 IU/ml, 1.30 IU/m and 13.18 IU/ml, 2.13 IU/ml, 1.87 IU/ml, 1.84 IU/m, respectively. There was no significant difference in the RVNA GMT levels at the same time point between the two regimen groups ( F=1.971, P=0.164). The RVNA GMT levels in the two regimen groups had the same trend of attenuation over time (time*group F=0.702, P=0.435). Retrospective study: The RVNA positive rates in 1-year, 2-year and 3-year after immunization in the Zagreb and Essen regimen group were 100%, 95%, 91.43%和94.73%, 86.21%, 87.5%, respectively. There was no statistically significant difference in the RVNA positive rates at the same time point between the two regimen groups ( P>0.05). The RVNA GMT in 1-year, 2-year, and 3-year groups after immunization in the Zagreb and Essen regimen group were 2.65 IU/ml, 2.03 IU/ml, 1.57 IU/ml和3.2 IU/ml, 2.58 IU/ml, 2.45 IU/ml, respectively. There was no significant difference in the RVNA GMT levels at the same time point between the two regimen groups ( P>0.05). Conclusions:The PVRV showed the same excellent immunogenicity and immune persistence after the vaccination with the Zagreb and Essen regimens.

OBJECTIVE: To explore the value of BACs-on-Beads (BoBs) for the practice of prenatal diagnosis. METHODS: The results of chromosomal karyotyping and BoBs of 1773 prenatal samples were compared. Microdeletions and microduplications detected by BoBs were subjected to chromosome microarray analysis (CMA) with informed consent from patients. RESULTS: BoBs has detected 46 cases of common aneuploidies involving chromosomes 13, 18, and 21, and 16 cases involving X and Y chromosomes. For 4 fetuses with normal results by BoBs, karyotyping analysis of amniotic fluid sample suggested low percentage mosaicisms (< 20%). BoBs has detected none of the 9 common microdeletions, but 14 male fetuses with Xp22 microdeletions and 5 with other microdeletions/microduplications. In 10 cases, the couples had chosen CMA verification, and the results were all consistent. CONCLUSION As a rapid diagnostic technique, BoBs has a high accuracy for common aneuploidies, and is capable of discovering certain chromosome microdeletions and microduplications. The difficulty lies in the inability to detect low proportion mosaicisms and the consultation following detection for male fetuses carrying Xp22 microdeletions.

Objective:To investigate gene basis of primary open angle glaucoma (POAG) by comparing gene expression profile of trabecular meshwork between POAG patients and normal controls by using RNA-sequencing.Methods:Trabecular meshwork specimen were obtained from trabeculectomy (POAG group, n=3) or donated eyes (control group, n=2). RNA was extracted and sequenced in both groups, gene expression profiles were analyzed and compared between them, and different expression genes associated with POAG were revealed by using Database for Annotation, Visualization and Integrated Discovery (DAVID) and Protein Analysis Through Evolutionary Relationships (PANTHER) gene list analysis.Written informed consent was obtained from each patient or the family members prior to entering the study cohort.The study protocol was approved by the Ethics Committee of Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong [No.EC20140311(3)-P01]. Results:(1)Total of 28 821 genes were obtained from RNA-sequencing, 22 genes were statistically significant between the two groups, of which one gene was up-regulated and 21 genes were down-regulated; (2)Genes that expressed differently had concentrated functions, biological process involved keratinization, epidermis development and intermediate filament cytoskeleton organization, cellular component related to keratin filament, intermediate filament, extracellular exosome and haptoglobin-hemoglobin complex, molecular function related to structural molecule activity and structural constituent of cytoskeleton; (3)Significantly enriched PANTHER pathways were plasminogen activating cascade, p38 MAPK pathway, oxidative stress response and p53 pathway.Conclusions:Trabecular meshwork and extracellular matrix remodeling due to abnormal keratin expression, structural change of intermediate filament cytoskeleton and misregulation of plasminogen activating cascade, p38 MAPK pathway were possible etiology of POAG.Differential expressed genes that related to POAG mainly involve cytoskeleton associated genes and extracellular matrix remodeling genes.Thus, regulation of these genes may have an effect on glaucomatous treatment.

Objective:To investigate the correlation between the muscle mass loss and severe postoperative pulmonary complications(PPC)in elderly patients with non-small cell lung cancer(NSCLC).Methods:Elderly patients with NSCLC undergoing lobectomy at the Lung Cancer Institute and the Department of Thoracic Surgery of Guangdong Provincial People's Hospital were recruited from Feb.2019 to Dec.2019.Data of the body composition, lung function, respiratory muscle strength test, cardiopulmonary exercise test were collected before operation.All patients were grouped into two groups: with versus without severe PPC at 30 d after operation.The differences of the above parameters were compared between the two groups.A multiple logistic regression analysis was used to analyze the risk factors for severe PPC.Results:In this study, 120 elderly NSCLC patients undergoing lobectomy were recruited, All evaluations were completed in 113 patients(aged 68.13±7.01 years)in whom, 21(18.58%, 21/113)patients had serious PPC.Compared with patients without PPC, patients with severe PPC had a lower appendicular skeletal muscle mass index(ASMI)(5.67±0.90 kg/m 2vs.7.71±1.40 kg/m 2, t=3.900, P=0.001), a lower forced expiratory volume in 1 second(FEV 1)(1.85±0.40 L vs.2.12±0.57 L, t=2.412, P=0.027), a lower maximal mid-expiratory flow(MMF)(1.40±0.69 L/s vs.2.11±1.09 L/s, t=2.502, P=0.021), a lower maximum inspiratory pressure(Pimax)(55.13±32.52 cmH 2O vs.64.71±20.60, t=0.778, P=0.047, 1 cmH 2O=0.098 kPa), a lower maximal oxygen consumption(Vo 2max)(1.14±0.41 L/min vs.1.40±0.34 L/min, t=0.779, P=0.046), a lower peak O 2 consumption(Vo 2max@kg)(20.00±1.91 L·min -1·kg -1vs.22.33±2.37 L·min -1·kg -1, t=0.813, P=0.041). Multiple logistic regression analysis showed that in addition to FEV 1( OR=2.824, 95% CI: 1.127-5.158, P=0.001)and Vo 2max@kg( OR=3.149, 95%CI: 1.829-6.592, P<0.001), ASMI was also an independent risk factor for serious PPC( OR=1.919, 95% CI: 1.604-3.466, P=0.006), in which the best cut-off value was 6.295 kg/m 2, the sensitivity and specificity were 0.816 and 0.818 respectively, and the area under the receiver operating characteristic(ROC)curve(AUC)was 0.887(95% CI: 0.793-0.981, P<0.0001). Conclusions:Muscle mass loss can increase the risk for the occurrence of severe PPC within 30 days after lobectomy in elderly patients with NSCLC.