Objective:To explore the expression of family with sequence similarity 83 member A (FAM83A) in colorectal cancer, and the effect of FAM83A knockdown on the proliferation of colorectal cancer cells and the related mechanism.Methods:The expression of FAM83A in the tissues of 102 patients with colorectal cancer and its adjacent tissues was detected by immunohistochemistry. HCT116 cells were divided into experimental group and control group. The experimental group cells were transfected with FAM83A-siRNA plasmid, and the control group cells were transfected with MOCK-siRNA plasmid. The mRNA content of FAM83A in each group was detected by fluorescence quantitative PCR. The expressions of FAM83A, P13K, p-AKT and p-mTOR in each group were detected by Western blot. CCK8 assay and clonogenesis assay were used to detect cell proliferation.Results:The positive rate of FAM83A in colorectal cancer patients was 88.23% (90 cases /102 cases), and the expression rate of FAM83A in paracancer tissues was 10.78% (11 cases /102 cases). The expression rate of Fam83a in colorectal cancer tissues was significantly higher than that in paracancer tissues, with statistical significance ( P<0.001). After siRNA transfection, the mRNA expression levels of FAM83A in HCT116 cells of the experimental group and control group were 1.23±0.20 and 0.43±0.12, respectively, and the protein expression levels of FAM83A were 1.19±0.11 and 0.23±0.08, respectively. The expression levels of P13K were 1.21±0.17 and 0.28±0.09, the expression levels of p-AKT were 1.35±0.23 and 0.57±0.18, and the expression levels of p-mTOR were 1.48±0.20 and 1.05±0.14. The expression of P13K, p-Akt and p-mTOR was down-regulated (all P<0.05). The absorbance of HCT116 cells in the experimental group and the control group was 1.09±0.22 and 2.21±0.27, respectively. The cloning rate of HCT116 cells in the experimental group and the control group was 21.6%±2.4% and 62.7%±4.1%, respectively. The proliferation ability of HCT116 cells in the experimental group decreased significantly ( P<0.05) . Conclusions:The expression of FAM83A is significantly increased in colorectal cancer tissues, which may be related to the malignant degree of colorectal cancer. FAM83A affects the proliferation of colorectal cancer cells through the P13K/AKT/mTOR signaling pathway.

Allergic asthma is a complex，polygenic disease characterized by chronic inflammation and airway hyperresponsiveness. Fungus and dust mites are the most important inhaled allergens of allergic asthma，and often exist in the form of mixed allergens. In recent years，genetic studies have shown that several genes are associated with allergic asthma attacks. This article reviews the studies on the genes related to allergic asthma caused by dust mites and fungus，such as a disintegrin and metalloproteinase 33（ADAM33），interleukin-4（IL-4），glycoprotein A repetitions predominant（GARP），toll like receptor 3（TLR3），mannose-binding lectin 2（MBL2），chemokine（C-C motif）ligand 17（CCL17）and other genes .

Objective To analyze the characteristics of work-related musculoskeletal disorders (WMSDs) among sonographers in Guangdong Province, and to explore the disease pattern of the cases. Methods A total of 512 sonographers from 31 hospitals in Guangdong Province were selected as the research subjects using stratified cluster sampling method. The prevalence of WMSDs in the past year was investigated using the Musculoskeletal Disorders Questionnaire, and the characteristics of WMSDs were analyzed. Latent class analysis was used to identify the disease pattern of WMSDs. Results The overall prevalence of WMSDs was 94.3%. The top five affected body parts were right shoulder, neck, right hand/wrist, lower back and right forearm/elbow, with the prevalence of 80.3%, 75.4%, 61.1%, 55.5% and 45.3%, respectively. The prevalence of WMSDs was higher on the right side for the shoulder, hand/wrist, forearm/elbow, hip/leg and knee compared with the left side (80.3% vs 31.3%, 61.1% vs 13.9%, 45.3% vs 10.0%, 17.4% vs 8.6%, 13.1% vs 8.4%, all P<0.05). The prevalence of WMSDs increased with work years, as well the prevalence of WMSDs in the top five affected body parts among the sonographers (all P<0.05). However, there were no statistical differences in prevalence of WMSDs between general hospitals and maternal and child health hospitals, tertiary hospitals and non-tertiary hospitals, Pearl River Delta hospitals and non-Pearl River Delta hospitals; there was also no statistical difference between different genders and age groups of the sonographers (all P>0.05). The best-fit latent disease pattern for sonographers WMSDs comprised three categories： symptom of neck-right shoulder, symptom of neck-lower back-right shoulder-right elbow-right hand/wrist, and symptom of multi-parts above the knees, with the latent probabilities of 0.438, 0.427 and 0.135, respectively. Conclusion The prevalence of WMSDs in sonographers is extremely high, with a dose-effect relationship with work years. The most common affected parts are neck, lower back and right shoulder, right hand/wrist, and right forearm/elbow. The prevalence of WMSDs in the right side of limb was higher than that in the left. WMSDs primarily occur in multiple parts simultaneously. The most common symptoms occur in the neck-right shoulder and neck-lower back-right shoulder-right elbow-right hand/wrist.

Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9), the third-generation genome editing tool, has been favored because of its high efficiency and clear system composition. In this technology, the introduced double-strand breaks (DSBs) are mainly repaired by non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathways. The high-fidelity HDR pathway is used for genome modification, which can introduce artificially controllable insertions, deletions, or substitutions carried by the donor templates. Although high-level knock-out can be easily achieved by NHEJ, accurate HDR-mediated knock-in remains a technical challenge. In most circumstances, although both alleles are broken by endonucleases, only one can be repaired by HDR, and the other one is usually recombined by NHEJ. For gene function studies or disease model establishment, biallelic editing to generate homozygous cell lines and homozygotes is needed to ensure consistent phenotypes. Thus, there is an urgent need for an efficient biallelic editing system. Here, we developed three pairs of integrated selection systems, where each of the two selection cassettes contained one drug-screening gene and one fluorescent marker. Flanked by homologous arms containing the mutated sequences, the selection cassettes were integrated into the target site, mediated by CRISPR/Cas9-induced HDR. Positively targeted cell clones were massively enriched by fluorescent microscopy after screening for drug resistance. We tested this novel method on the amyloid precursor protein (APP) and presenilin 1 (PSEN1) loci and demonstrated up to 82.0% biallelic editing efficiency after optimization. Our results indicate that this strategy can provide a new efficient approach for biallelic editing and lay a foundation for establishment of an easier and more efficient disease model.
Alleles ; CRISPR-Cas Systems ; DNA End-Joining Repair ; Gene Editing/methods* ; Recombinational DNA Repair

Objective:To evaluate the positive rate of the bronchodilation test (BDT) in asthmatic children with normal forced expiratory volume in 1 second (FEV 1), forced vital capacity (FVC) and FEV 1/FVC, so as to improve the recognition of the importance of the BDT test in asthmatic children with normal FEV 1, FVC and FEV 1/FVC. Methods:Children aged 5-14 who were diagnosed with asthma in the outpatient clinic of Shandong Provincial Hospital Affiliated to Shandong University from September 2018 to August 2019 and willing to receive pulmonary function and BDT examinations were enrolled.Data of pulmonary function of children with normal FEV 1, FVC and FEV 1/FVC were collected to analyze the rate of positive BDT results and the status of small airway function. Results:A total of 1 631 asthmatic children with normal FEV 1, FVC and FEV 1/FVC were enrolled in this study, including 1 414 children with normal pulmonary function and 217 children with small airway dysfunction.Fifteen minutes after the bronchodilator was inhaled, 127 children (87 males and 40 females) showed positive BDT results, accounting for 7.8%.Among these children, 62 cases (28.6%) with co-existing small airway dysfunction showed positive BDT results.The improvement rate of FEV 1 was 8.0% to 11.9% in 132 cases (8.1%). The FEV 1 before bronchodilator inhalation accounted for (98.5±10.3)% of the predicted value.Fifteen minutes after terbutaline sulfate inhalation, the improvement rate was 13.5% (12.5%, 16.2%). The improvement rates of forced expiratory flow at 50% of FVC exhaled (FEF 50, r=-0.339, P<0.01), forced expiratory flow at 75% of forced vital capacity exhaled (FEF 75, r=-0.400, P<0.01), maximum mid-expiratory flow(MMEF, r=-0.375, P<0.01) were negatively correlated with their baseline values.The improvement rate of FEV 1 was not associated with its baseline value ( r=-0.128, P=0.153), but negatively correlated with the baseline value of MMEF ( r=-0.231, P<0.01). Conclusions:BDT results are positive in some asthmatic children with normal FEV 1, FVC and FEV 1/FVC.It is recommended that BDT testing should be conducted as much as possible in the diagnosis and follow-up of children with typical or atypical asthma.In this way, the diagnosis can be confirmed and the current optimal results can be obtained.Meanwhile, small airway function testing is helpful for comprehensive assessment of asthma and its control level.

Objective:To investigate the clinical significance of MRI dynamic enhancement in defining the upper edge of adenocarcinoma of esophagogastric junction.Methods:The clinical data of 73 patients with adenocarcinoma on the esophagogastric junction operated from Jul 2018 to Aug 2019 in the Affiliated Cancer Hospital of Zhengzhou University were retrospectively analyzed. All patients underwent MRI examination within one week before surgery. First, the T 2WI, diffusion-weighted imaging and dynamic enhanced images of each patient were carefully observed to define the tumor location, size, shape, signal and enhancement mode, then the distances between the upper edge of the tumor and the cardiac incisure on MRI dynamic enhancement were measured independently, and compared with the measured distance in surgical fresh specimens. Results:The mean location of tumor upper edge measured in MRI of 73 patients was (1.75±1.98)cm, while measured in the surgical specimen was (1.72±1.97)cm. There was no significant difference between the two groups ( t=0.572, P=0.569). The intraclass correlation efficient between the two groups was excellent (ICC=0.974, 95% CI: 0.959-0.984, P<0.01). Conclusion:The measurement result of tumor upper edge in MRI is basically consistent with that of surgical specimens. MRI can be used to locate the tumor upper edge for adenocarcinoma of esophagogastric junction before operation.

Objective:To investigate the normal range of exhaled nitric oxide in healthy children aged 6-18 in Jinan.Methods:The healthy school children aged 6-18 in Jinan from October 11 to 26, 2017 were selected for questionnaire survey, physical examination and exhaled nitric oxide test.The levels of mouth exhaled nitric oxide at the flow rate of 50 mL/s (FeNO 50) and mouth exhaled nitric oxide at the flow rate of 200 mL/s(FeNO 200), alveolar nitric oxide (CaNO), and nasal exhaled nitric oxide at the flow rate of 10 mL/s(FnNO 10) were measured by the electroche-mical method.The distributions of FeNO 50, FeNO 200, CaNO and FnNO 10 were analyzed, and their correlations with gender, age, height and body mass index (BMI) were discussed by the multiple linear regression model. Results:A total of 772 healthy children were enrolled in this study, including 364 males and 408 females, with a median age of 12.1(11.8-12.3) years old, a median height of 154.8(153.6-156.0) cm, and a median BMI of 20.3 (20.0-20.6) kg/m 2. The measured values of FeNO 50, FeNO 200, CaNO and FnNO 10 fluctuated in the range of 3.0-168.0 ppb, 2.0-44.0 ppb, 0.5-44.2 ppb and 0-1 253.0 ppb, respectively.FeNO 50, FeNO 200 and CaNO values showed skewed a distribution, and their 95% upper limits were 35.0 ppb, 13.3 ppb and 8.5 ppb, respectively.The geometric mean(95% CI) of FeNO 50 in males (95% CI)[14.6 (13.7-15.5) ppb] was significantly higher than that in females [13.3(12.7-14.0) ppb], and the difference was statistically significant ( Z=1.470, P=0.027). The multiple linear regression results suggested that, FeNO 50 was positively correlated with age and height ( β=0.023, 0.007, respectively, all P<0.05), and negatively correlated with BMI ( β=-0.016, P<0.05). The geometric mean (95% CI) of FeNO 200in males[7.1 (6.8-7.4) ppb] was significantly higher than that in females[6.4 (6.1-6.6) ppb], and the difference was statistically significant( Z=1.747, P=0.004). The multiple linear regression results suggested that, FeNO 200 was positively correlated with height ( β=0.005) and negatively correlated with gender(female β=-1.126) (all P<0.05). There was no significant difference between male and female in CaNO, which had no correlation with gender, age, height and BMI (all P>0.05). FnNO 10 showed a normal distribution, with a mean value of 456.2 ppb, 95% CI of 29.3-863.4 ppb.The geometric mean (95% CI) of FnNO 10 in males [408.7 (377.1-443.0) ppb] was significantly higher than that in females [368.8 (339.0-401.3) ppb], and the difference was statistically significant ( Z=1.722, P=0.005). The multiple linear regression results indicated that FnNO 10 was related to gender ( β=-36.098, P<0.05), and not correlated with age, height and BMI (all P>0.05). Conclusions:The normal ranges of FeNO 50, FeNO 200, CaNO and FnNO 10 in healthy children aged 6-18 in Jinan are 3.0-35.0 ppb, 2.0-13.3 ppb, 0.5~8.5 ppb and 29.3-863.4 ppb, respectively.FeNO 50 is correlated with age, height and BMI.FeNO 200 is correlated with gender and height.CaNO and FnNO 10 are not correlated with age, height or BMI.

Objective:To investigate the normal range of exhaled nitric oxide (FeNO) in 6-18-year-old children in China, so as to provide a data base for the establishment of FeNO standards for Chinese children.Methods:A multi-center study was conducted on 5 949 children aged 6-18 (3 101 males and 2 848 females) in 16 pro-vinces of 7 administrative districts in China.According to the technical standard recommended by American Thoracic Society/European Respiratory Association, FeNO was measured, and the relationship of FeNO with the sex, age, height, weight, body mass index and region was discussed.Results:The geometric mean FeNO value of Chinese children aged 6-18 was 14.1 ppb, and its 95% confidence interval (skewness distribution) was 1.0-38.2 ppb.The geometric mean FeNO values of children aged 6-11 and 12-18 were 13.1 ppb and 15.7 ppb, respectively, and their 95% confidence intervals (skewness distribution) were 1.0-38.1 ppb and 2.0-38.2 ppb.For children at and under 11 years old, FeNO decreased with age, with a mean decline of 1 ppb per year.The multiple linear regression results suggested that there was a significant correlation between FeNO and age for children aged 6-11, and FeNO of children aged 12-18 was significantly correlated with the gender, height, and region(all P<0.01). Conclusions:FeNO values of Chinese children and adolescents in this study are higher than those obtained by the previous study conducted from 2010 to 2012.For children aged 12-18, 16 ppb is recommended as the clinical cut-off point.For children at or under 11 years old, the influence of age on FeNO should be considered, and the cut-off point of FeNO decreases by 1 ppb as the age is reduced by one year.

Objective To investigate the correlation between quantitative parameters of dynamic contrast?enhanced MRI (DCE?MRI) after neoadjuvant chemotherapy and pathological grades in esophageal squamous cell carcinoma. Methods Fifty?six patients with esophageal squamous cell carcinoma who were confirmed by esophagoscope and received neoadjuvant chemotherapy before operation between September 2015 and December 2017 in the Affiliated Cancer Hospital of Zhengzhou University were prospectively analyzed, and MRI examination was performed within one week before operation. All patients underwent routine chest MRI and DCE?MRI scanning, and quantitative parameters of DCE?MRI, including volume transfer constant (Ktrans),exchange rate constant (Kep) and extravascular extracellular volume fraction (Ve) were measured. Pathological grading was assessed as highly differentiated, moderately differentiated, poorly differentiated,and undifferentiated. Intraclass correlation coefficient (ICC) was calculated from the results of two radiologists. Kruskal?Wallis H test was used to compare the differences of quantitative parameters between different pathological grade groups of DCE?MRI,and Mann?Whitney U test was utilized to compare the intraclass differences among pathological grades. Spearman rank correlation analysis was performed for evaluating the correlation between DCE?MRI parameters and pathological grade of esophageal squamous cell carcinoma. The receiver operating characteristic (ROC) curves were used to evaluate the diagnosis accuracy of different DCE?MRI parameters in pathological grade of esophageal squamous cell carcinoma after neoadjuvant chemotherapy. Results The 56 patients were divided into four groups according to pathological findings: well differentiated group (n=8), moderately differentiated group (n=39), poorly differentiated group (n=9) and undifferentiated group (n=0). The differences of Ktransmean,Ktrans75%,Kepmax, Kepmean,Kep75% between different pathological grading groups were statistically significant (all P<0.05),and these parameters showed positive correlation significantly with pathological grading (r values were 0.778, 0.632, 0.594, 0.725, 0.489 respectively, all P<0.05). The ROC curve area of Ktransmean, Ktrans75% in the diagnosis of pathological grade for esophageal squamous cell carcinoma was 0.750,0.856,respectively. The diagnostic efficiency of Ktrans75% was the best with the diagnostic threshold of 0.693/min,sensitivity of 87.5%, specificity of 78.5%, respectively. Conclusion The quantitative parameters of DCE?MRI after neoadjuvant chemotherapy in esophageal squamous cell carcinoma have the potential value for predicting pathological grade.

Objective To investigate the detection and distribution of hospitalized specimens from a tertiary hospital over 5 years. Methods Specimens of sputum, urine, blood, secretions and puncture fluid were collected from patients admitted to the Harrison International Peace Hospital from November 2013 to November 2018. The origin of specimens, the distribution of departments and the distribution of pathogenic bacteria isolated were analyzed retrospectively. Results A total of 61 286 specimens were sent for examination during the 5 years. The top 5 specimens were sputum culture (n = 18 302, 29.9%), sputum smear (n = 11 253, 18.4%), blood culture (n = 9 713, 15.8%), urine culture (n = 6 448, 10.5%) and secretion culture (n = 6 133, 10.0%), accounting for 84.6% (51 849/61 286). Sputum specimens accounted for 48.2% (29 555/61 286) with the largest proportion. The number of specimens from medical wards was much higher than that from surgical wards (specimens: 25 468 vs. 10 521), respiratory medicine, department of critical care medicine and emergency intensive care unit (EICU) were important sources of pathogenic specimens in the hospital, accounting for 29.8% (18 243/61 286) in total. The average positive rate of all specimens was 23.5% (14 424/61 286). The positive rates of sputum culture and urine culture were 29.7% (5 428/18 302) and 35.4% (2 281/6 448), respectively, while the positive rate of blood culture was only 6.6% (643/9 713). Escherichia coli was the most common pathogen in all specimens except for sputum culture and fecal culture. Escherichia coli [40.6% (926/2 281)], Klebsiella pneumoniae [9.2% (210/2 281)], Pseudomonas aeruginosa [8.2% (188/2 281)], Enterococcus faecalis (group D) [6.6% (151/2 281)] and Candida albicans [3.2% (73/2 281)] were the most common pathogens in urine culture. Klebsiella pneumoniae [24.1% (1 309/5 428)], Acinetobacter baumannii [21.3% (1 154/5 428)], Pseudomonas aeruginosa [15.1% (818/5 428)], Escherichia coli [6.5% (351/5 428)] and Maltose oligotrophomonas maltose [5.8% (316/5 428)] were the most common pathogens in sputum culture. Escherichia coli [36.5% (235/643)], Klebsiella pneumoniae [10.9% (70/643)], Pseudomonas aeruginosa [4.8% (31/643)], Staphylococcus epidermidis [3.4% (22/643)] and Staphylococcus humanis [3.3% (21/643)] were the most common pathogens in blood culture. Conclusion Specimens sent for examination by inpatients are mainly from internal medicine wards, mainly from sputum, blood and urine, and the detected pathogens are mainly Gram-negative bacteria.