Objective To investigate the effect of miR-142-3p on the apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1.Methods AR42J cells were divided into blank group(blank),acute pancreatitis model group(AP,100 nmol/L cerulein for 24 h),and then transfected with miR-142-3p mimics,mimics NC,miR-142-3p inhibitor and inhibitor NC,respectively.The cells in the model group were recorded as miR-142-3p mimics group,mimics NC group,miR-142-3p inhibitor group and inhibitor NC.The expression of miR-142-3p in cells was detected by RT-qPCR.The protein expressions of HMGB1,caspase-3,Bax and Bcl-2 were detected by Western blot.Hoechst staining was used to determine cell apoptosis.The apoptosis rate of cells was detected by flow cytometry.The targeting relationship between miR-142-3p and Hmgb1 was determined by dual luciferase reporter gene assay.Results Compared with blank control group,the expression level of miR-142-3p in the AP group was significantly down-regulated(P＜0.01),the expression level of HMGB1 and caspase-3 proteins was up-regulated(P＜0.05),the expression level of Bax protein was significantly up-regulated(P＜0.01),the expression level of Bcl-2 protein was significantly decreased(P＜0.01)and the apoptosis rate increased significantly(P＜0.01).Compared with the mimics NC group,the level of miR-142-3p in the miR-142-3p mimics group was significantly up-regulated(P＜0.01),the expression of HMGB,caspase-3 and Bax proteins was significantly down-regulated(P＜0.01),the expression of Bcl-2 protein was up-regulated(P＜0.05),and the apoptosis rate decreased signifi-cantly(P＜0.01).Compared with inhibitor NC group,the expression level of miR-142-3p in miR-142-3p inhibitor group was down-regulated(P＜0.05),the expression levels of HMGB1,caspase-3 and Bax proteins were signifi-cantly up-regulated(P＜0.01),the expression level of Bcl-2 protein was decreased(P＜0.05)and the apoptosis rate increased significantly(P＜0.01).The dual luciferase reporter gene assay showed that Hmgb1 was the target gene of miR-142-3p.Conclusions 1)The expression of miR-142-3p was low in the model group.2)miR-142-3p can inhibit the apoptosis of AR42J cells by inhibiting the expression of Hmgb1.

Objective: To investigate the effects of fiber surface deposited with silicon dioxide films by atomic layer deposition on properties of dental fiber-reinforced composites. Methods : SiO2 films were deposited on the surface of quartz fiber by atomic layer deposition(ALD). Then the quartz fiber was used to manufacture fiber resin composites, which were divided into four groups: A(no soaking agent removal), B(soaking agent removed), C(soaking agent removed and silanization), and D(soaking agent removed, 600 ALD cycles performed and then silanization). Scanning electron microscopy, water contact angle test, hygroscopicity test, CCK8 test and three-point bending test were used to investigate the properties of fiber resin composites. Results: The surface morphology of the quartz fiber treated by ALD was smooth and had no obvious change compared with that before treatment. Moreover, the quartz fiber showed hydrophobicity after silanization. The results of three-point bending test revealed that the mechanical properties of fiber-resin composites modified by ALD were significantly improved(P<0.05). When viewed by scanning electron microscopy, a good interfacial bonding could be seen between quartz fibers and the resin matrix in Group D. In addition, it was found that Group D had low absorbability, low solubility and good biocompatibility. Conclusion It is shown that deposition of SiO2 films on the quartz fiber by ALD can significantly enhance the mechanical properties of fiber-reinforced composites.

Primary ciliary dyskinesia (PCD) is a congenital, motile ciliopathy with pleiotropic symptoms. Although nearly 50 causative genes have been identified, they only account for approximately 70% of definitive PCD cases. Dynein axonemal heavy chain 10 (DNAH10) encodes a subunit of the inner arm dynein heavy chain in motile cilia and sperm flagella. Based on the common axoneme structure of motile cilia and sperm flagella, DNAH10 variants are likely to cause PCD. Using exome sequencing, we identified a novel DNAH10 homozygous variant (c.589C > T, p.R197W) in a patient with PCD from a consanguineous family. The patient manifested sinusitis, bronchiectasis, situs inversus, and asthenoteratozoospermia. Immunostaining analysis showed the absence of DNAH10 and DNALI1 in the respiratory cilia, and transmission electron microscopy revealed strikingly disordered axoneme 9+2 architecture and inner dynein arm defects in the respiratory cilia and sperm flagella. Subsequently, animal models of Dnah10-knockin mice harboring missense variants and Dnah10-knockout mice recapitulated the phenotypes of PCD, including chronic respiratory infection, male infertility, and hydrocephalus. To the best of our knowledge, this study is the first to report DNAH10 deficiency related to PCD in human and mouse models, which suggests that DNAH10 recessive mutation is causative of PCD.
Humans ; Male ; Animals ; Mice ; Semen/metabolism* ; Dyneins/metabolism* ; Cilia/metabolism* ; Mutation ; Ciliary Motility Disorders/genetics*

OBJECTIVE: To determine the carrier rate for 21 inherited metabolic diseases among a Chinese population of childbearing age. METHODS: A total of 897 unrelated healthy individuals (including 143 couples) were recruited, and DNA was extracted from their peripheral blood samples. Whole exome sequencing (WES) was carried out to screen potential variants among 54 genes associated with 21 inherited metabolic diseases. Pathogenic and likely pathogenic variants and unreported loss-of-function variants were analyzed. RESULTS: One hundred fourty types of pathogenic/likely pathogenic variants (with an overall number of 183) and unreported loss-of-function variants were detected, which yield a frequency of 0.20 per capita. A husband and wife were both found to carry pathogenic variants of the SLC25A13 gene and have given birth to a healthy baby with the aid of preimplantation genetic diagnosis. The detected variants have involved 40 genes, with the most common ones including ATP7B, SLC25A13, PAH, CBS and MMACHC. Based on the Hardy-Weinberg equilibrium, the incidence of the 21 inherited metabolic diseases in the population was approximately 1/1100, with the five diseases with higher incidence including citrullinemia, methylmalonic acidemia, Wilson disease, glycogen storage disease, and phenylketonuria. CONCLUSION This study has preliminarily determined the carrier rate and incidence of 21 inherited metabolic diseases among a Chinese population of childbearing age, which has provided valuable information for the design of neonatal screening program for inherited metabolic diseases. Pre-conception carrier screening can provide an important measure for the prevention of transmission of Mendelian disorders in the population.
Asians/genetics* ; China ; Exome ; Female ; Humans ; Infant, Newborn ; Metabolic Diseases/genetics* ; Mitochondrial Membrane Transport Proteins/genetics* ; Oxidoreductases/genetics* ; Whole Exome Sequencing

Objective To investigate the effect of naringenin on the killing rate of natural killer (NK) cells and related mechanism by amplification of human peripheral blood mononuclear cells into NK cells in vitro and co-culture with hepatocellular carcinoma (HCC) CLC5 cells at a ratio of 1∶ 1. Methods A lymphocyte separation medium was used to isolate human peripheral blood mononuclear cells, which were induced with recombinant human interleukin-2 in vitro to culture NK cells. CCK-8 assay was used to measure the proliferation of HCC cells after human HCC cells were treated with naringenin (0, 3.125, 6.25, 12.5, 25, and 50 μmol/L) for 0, 24, and 48 hours, and after human NK cells were treated with different concentrations of naringenin for 24 hours, CCK-8 assay was used to measure the proliferation of NK cells. CellTiter-LumiTM was used to measure the killing rate of NK cells after the NK-HCC cell co-culture system at the ratio of 1∶ 1 was treated with naringenin for 24 hours. Quantitative real-time PCR was used to measure the gene expression of the activating receptor NKG2D in NK cells and NKG2D ligands in HCC cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results After being induced and cultured by recombinant human interleukin-2, NK cells were amplified to 82.33%±0.70% of human peripheral blood mononuclear cells. After naringenin treatment for 24 hours, there was no significant difference in the proliferation rate of HCC CLC5 cells between all mass concentration groups (all P > 0.05), and in the 25 and 50 μmol/L mass concentration groups, naringenin significantly promoted the proliferation of NK cells (both P < 0.000 1). After the NK-HCC cell co-culture system at the ratio of 1∶ 1 was treated with naringenin for 24 hours, there was a significant increase in the killing rate of NK cells in the 25 and 50 μmol/L mass concentration groups (both P < 0.000 1). After the co-culture system was treated with naringenin for 24 hours, naringenin had no effect on the expression of NKG2D in NK cells in the 25 and 50 μmol/L mass concentration groups, and it also had no effect on the expression of MICB and ULBP2 in HCC cells (all P > 0.05); it significantly upregulated the expression of the NKG2D ligands such as ULBP1 and ULBP3 in HCC cells (all P < 0.001). Conclusion Naringenin may increase the killing activity of NK cells by upregulating the expression of NKG2D ligands in HCC cells.

Nonalcoholic fatty liver disease has become the most important chronic liver disease in China and the leading cause of abnormal liver biochemical parameters in physical examination. Mitophagy can degrade the mitochondria with impaired function and misfolded protein in the liver to regulate cell death and help to maintain stable lipid metabolism in the liver. The binding of adiponectin secreted by adipose tissue to adiponectin receptor 2 can increase the secretion of autophagy agonists in Kupffer cells, induce hepatocyte mitophagy, promote lipid metabolism, and thus reduce liver inflammation, which provides new thoughts for the research and development of drugs with a marked clinical effect and a clear mechanism of action in the treatment of nonalcoholic fatty liver disease.

OBJECTIVE: To optimize the condition for chromosome flaking of mesenchymal stem cells to ensure the cytogenetic quality control of expanding production and clinical application. METHODS: Chromosomal flaking methods were optimized from current chromosome preparation techniques from the aspects of MSCs cell culture concentration, colchicine treatment time and low permeability time. RESULTS: By repeated pre-experiments, the optimal MSCS chromosome flaking condition of MSCs was determined as cell culture concentration of (1-2)× 10 cells per T25 cell culture bottle, and the colchicines processing time was determined as 2 hours and 10 minutes, and the low permeability was 1 hour. CONCLUSION The optimized chromosome flaking condition can fulfill the requirement of cytogenetic quality control for MSCs.
Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Chromosome Disorders ; Chromosomes, Human ; Cytogenetics ; Humans ; Mesenchymal Stem Cells

Portal hypertension refers to a series of clinical manifestations caused by elevated pressure of the portal vein system, which can cause portal hypertension by causing portal venous obstruction and / or increased blood flow. A typical clinical manifestation in patients with decompensated cirrhosis is portal hypertension. A severe complication of portal hypertension is esophagogastric varices bleeding, refractory ascites, and hepatic encephalopathy. The effective reduction of portal pressure can reduce the incidence of complications, improve the prognosis and reduce the mortality. At present, the commonly used clinical methods for reducing portal hypertension include drug therapy, minimally invasive interventions, surgical treatment, and liver transplantation. This article reviews the current status of integrated traditional Chinese and Western medicine for portal hypertension.

Objective To investigate the successful rate and the therapeutic effect of loosening posterior tubercle of cervical transverse process with CT-guided insertion or blind insertion of small needle knife for cervical spondylotic radiculopathy. Methods Ninety patients with cervical spondylotic radiculopathy were divided into CT-guided insertion group and blind insertion group in the admission order using the random number table method, 45 cases in each group. CT-guided insertion group adopted the insertion of small needle knife following by the guide of CT, and the blind insertion group adopted the insertion of small needle knife following by cervical bony landmarks and muscles. After insertion, the position of the small needle knife in patients of the two groups was located by CT scan. If the needle knife did not arrive to the foci, it will be inserted again following by the guide of CT till successful insertion and location. The successful rate for the two times of insertion was calculated, and the pain scores were evaluated by visual analogue scale(VAS). Results(1) The successful rate for the first insertion of CT-guided insertion group was 73.3％, and that of the blind insertion group was 47.8％, the difference being significant(P < 0.01). The successful rate for the second insertion of CT-guided insertion group was 92.2％, and that of the blind insertion group was 80.0％, the difference being significant (P<0.01). (2) After treatment for one week, pain VAS scores of the two group were decreased (P < 0.01 compared with those before treatment) , and the decrease in the CT-guided insertion group was more obvious than the blind insertion group (P < 0.05). Conclusion CT-guided insertion makes the small needle knife therapy be more accurate and safer, ensures much better therapeutic effect, and brings less pain in the patients.

OBJECTIVETo investigate the phenotype-genotype association of isodicentromere Y chromosome by analysis of two female patients carrying the chromosome with sexual development disorders.

METHODSThe karyotypes of the two patients were determined by application of conventional G banding of peripheral blood samples and fluorescence in situ hybridization (FISH). PCR was applied to detect the presence of SRY gene.

RESULTSConventional karyotype analysis showed case 1 to be a mosaic: mos.45,X[38]/46,X,+mar[151]/47,XY,+mar[5]/47,X,+mar × 2[2]/46,XY[4], FISH showed that 12 different cell lines were presented in the karyotype of case 1 and partial cell lines with SRY gene, the marker is an isodicentromere Y chromosome [idic(Y)(p)]. No mutation was found in the SRY gene. The karyotype of case 2 was mos.45,X[25]/46,X,+mar[35]. FISH showed the marker to be an idic(Y)(p) without the SRY gene.

CONCLUSIONThe karyotype of patients carrying idic(Y)(p) seems unstable, and female patients have the characteristics of short stature and secondary sexual hypoplasia. Karyotype analysis combined with FISH analysis can accurately determine the breakpoint of idic(Y) and identify the types of complex mosaic, which may facilitate genetic counseling and prognosis.

Adolescent ; Child ; Chromosomes, Human, Y ; Disorders of Sex Development ; genetics ; Female ; Humans ; Karyotype ; Sex Chromosome Aberrations ; Sex-Determining Region Y Protein ; genetics