Objective To investigate the effects of the rhein on the mitochondria fission and epithelial-mesenchymal transition(EMT)of breast cancer cells MDA-MB-231.Methods Human breast cancer cells were intervened with rhein,and the cells were divided into control group(0 μmol·L-1),low dose rhein group(100 μmol·L-1),and high dose rhein group(200 μmol·L-1).The proliferation activity of the cells was detected by CCK-8,and migrations was detected by Scratch-healing migration assay.The morphology and distribution of mitochondria were detected by transmission electron microscope,and the expression levels of Dynamin-related protein 1(Drp1),mitofusin2(Mfn2),E-cadherin,Vimentin proteins were detected by Western blot.Results Compared with control group,Rhein significantly reduced the protein expression of Drp1、Vimentin(P<0.05),and increased E-cadherin and Mfn2,thus down-regulating mitochondria fission,inhibiting cell proliferation and migration.High dose Rhein was better than low dose.Conclusion Rhein can inhibit the proliferation and migration of breast cancer cells by reducing the expression of Drp1,Vimentin and up-regulating Mfn2,E-cadherin proteins.

DECIDE-Diet trial was taken as a case to introduce the methods of blinding and blinding assessment for feeding trials, report the details of blinding, conduct a blinding survey and calculate Jame's BI and Bang's BI. Jame's BI was 0.683 (95% CI: 0.593~0.772). The Bang's BI for the intervention group was 0.340 (95% CI: 0.199~0.481), and for the control group was 0.086 (95% CI: -0.060~0.231). The blinding of the DECIDE)-Diet was generally successful, but the intervention group may infer their group to a certain extent. Feeding trials should report the details of blinding and consider blinding assessment.

Objective:Cushing′s disease(CD) is caused by the pituitary adrenocorticotroph hormone(ACTH) secreting adenomas, leading to increased serum cortisol levels and various abnormal metabolic processes. Untreated CD is linked to high mortality, thus it is critical to elucidate its pathogenesis. This study aims to explore the pathogenesis of pituitary ACTH adenomas using whole-genome sequencing analysis.Methods:Fresh tumor tissues and peripheral blood samples were collected in 9 confirmed cases of pituitary ACTH adenomas who underwent surgery. Whole genome sequencing was then performed, followed by analysis and verification of single nucleotide mutations, copy number variation(CNV) and chromosome structure variations.Results:Somatic USP8 mutations(p.Ser718del, p. Ser718Pro, p. Pro720Arg, p. Pro720Gln) were found in 5 patients, with a rate of 55.6%; CNV of USP8 was detected in 1 patient; TP53(p.Cys135Tyr), NF1(p.Val1049Glufs*11) and KMT2C(c.3323+ 1G>A) mutations were identified in 1 patient harboring wild-type USP8. CNV analysis showed a loss of heterozygosity in multiple chromosomes in a wild-type USP8 patient. Structural variations were found in 2 with unknown significance. No germline gene mutations were detected in this study.Conclusion:Somatic USP8 mutations, increased copy number of USP8, variations of tumor-related genes such as TP53 and extensive somatic CNV all contribute to pathogenesis of CD. Chromosomal structure variations may suggest high-risk pituitary ACTH adenomas, and call for frequent follow-up and aggressive treatment.

Objective:To summarize the prenatal sonographic characteristics of affected joints of fetal arthrogryposis multiplex congenita (AMC) by comparing the ultrasonographic features and the postnatal pathological manifestations.Methods:The cases of AMC detected by antenatal ultrasound and confirmed by postnatal pathology were collected in the First Affiliated Hospital of Sun Yat-Sen University and Kaiping Central Hospital from January 2015 to June 2020. The differences between prenatal ultrasonic manifestations, types of affected joints and postnatal pathological features were analyzed. And the different involvements of joints in AMC cases with or without other system abnormalities were also explored separately.Results:A total of 31 cases of AMC were included, in which 11 cases were with other system abnormalities and 20 cases without. No significant difference was observed in number of affected joints between these two groups ( P>0.05). The prenatal sonogram features were completely consistent with the postnatal pathological manifestations in 21 (21/31, 67.7%) cases. Among 31 cases, the involvement rates of joints were: interphalangeal joints of fingers (23/31, 74.2%), knee joints (20/31, 64.5%) and ankle joints (19/31, 61.3%), temporomandibular joint (11/31, 35.5%), wrists (11/31, 35.5%), elbow joints (10/31, 32.3%), interphalangeal joints of toes (6/31, 19.4%), spinal joints (2/31, 6.5%), shoulder joint (1/31, 3.2%) and hip joint (1/31, 3.2%), respectively. The coincidence rates of prenatal ultrasound in involved joints were: interphalangeal joints of fingers (100%), ankles (100%), spines (100%), hips (100%), wrists (90.9%), knees (75.0%), elbows (70.0%), jaws (54.5%), interphalangeal joints of toes (50.0%), and shoulders (0), respectively. Conclusions:When postural abnormalities of fetal upper and lower extremities are detected by prenatal ultrasound screening, especially overlapping fingers, extended knee and club foot, AMC should be kept on alert. Simultaneously, other joints should be carefully scanned to improve the prenatal detection rate of AMC.

Objective:To explore the effect of Tongxie-Yaofang on visceral sensitivity in IBS-D and the possible mechanism. Methods:Divided 30 male SD rats (one-day old) into normal group (10 rats) and IBS-D model group (20 rats) randomly. IBS-D was induced by the method of neonatal maternal separation and restraint stress. After successful modeling, the IBS-D model group was randomly divided into model group and Tongxie-Yaofang group, with 10 rats in each group. Tongxie-Yaofang group was given Tongxie-Yaofang formula, 4.92 g/ml by gavage, while the normal and model groups were given the same amount of normal saline, rats were gavaged with 2 ml/100 g body weight once a day for 14 days. The electromyography of the exorectus muscle was used to meature colorectal distension and by using electronic constant pressure apparatus to evaluate visceral sensitivity. The morphology of colon by HE staining and Enzyme-linked immunosorbent assay (ELISA) were used to determine the level of colonic 5-hydroxytryptamine (5-HT), qPCR was used to detect the colonic mRNA expression of serotonin transporter (SERT) and Western blot was used to detect SERT expression in colon and hypothalamus. Results:Compared with the model group, at the expansion pressure of 60 mmHg and 80 mmHg, the electromyographic response [(179.51 ± 18.26)% vs. (226.42 ± 25.78)%; (242.13 ± 15.42)% vs. (306.02 ± 51.51)%] in Tongxie-Yaofang group was significantly decreased ( P<0.05 or P<0.01). The colonic content of 5-HT was significantly lower than that in the model group [(8.85 ± 0.53) ng/mg vs.(12.25 ± 1.95) ng/mg] ( P<0.01), the expression of SERT mRNA (0.85 ± 0.12 vs. 0.38 ± 0.02) and SERT protein (0.53 ± 0.11 vs. 0.36 ± 0.17) in the colon was significantly increased ( P<0.05 or P<0.01), the expression of SERT protein (0.88 ± 0.12 vs. 0.36 ± 0.13) in the hypothalamus was significantly increased ( P<0.05). Conclusion:Tongxie-Yaofang could relieve the visceral hypersensitivity, which may be achieved by regulating the 5-HT and SERT expression.

OBJECTIVETo investigate the association of mutations in the hepatitis B virus (HBV) X gene (HBX, encoding the HBx protein) and development of hepatocellular carcinoma (HCC).

METHODSForty-four patients with HBV-related HCC participated in the study, along with 76 patients with chronic HBV infection who assessed as controls. All patients had serum HBV DNA levels that were higher than 10(3) copies/ml. Extracted HBV DNA was subjected to nested PCR to amplify the HBX gene, followed by direct sequencing. All sequencing data were compared to the consensus HBV sequence to identify mutations. The sequencing data were analyzed by Chromas and SeqMan software.

RESULTSMutations of G1467C, G/C1479A, C1485T and C1653T in the X region were found, but did not show any significant difference in occurrence between the HCC group and the chronic HBV infection group (P>0.05). The T1674C mutation in the X region, however, occurred more frequently in the HCC group (29.27% vs.6.67%, P<0.05). Prevalence of the T1753C mutation and the A1762T/G1764A double mutation in the BCP region was significantly higher in the HCC group than in the chronic HBV infection group (P<0.05) and in the group of patients with hepatitis B e antigen (HBeAg)-negative status compared to the patients with HBeAg-positive status (P<0.05).

CONCLUSIONIncidence of the T1674C mutation in the X region and of the T1753C mutation and the A1762T/G1764A double mutation in the BCP region was higher for patients with HBV-related HCC; the T1753C mutation and the A1762T/G1764A double mutation may inhibit the formation of HBeAg.

China ; Counseling ; Female ; HIV Infections ; diagnosis ; Humans ; Pregnancy

Induced pluripotent stem cells can differentiate into a variety of cell types,which promote the development of human disease model,drug toxicity screening and sources of autologous cells.However,there have been many problems in the induced pluripotent stem cells reprogramming,such as safety and low efficiency.Small molecules are considered as a promising method to improve the reprogramming processes of induced pluripotent stem cell,and more and more small molecules have been identified to maintain stem cell self-renewal,providing a new approach to produce the desired reprogramming cells.

Objective To investigate the relationship between the genotype /sub -genotype and antiviral treatment.Methods The clinical data of 159 patients with chronic HBV infections were collected,who were treated by adefovir dipivoxil (10mg/d)or entecavir(0.5mg/d)according to the disease′s conditions.The genotypes and subtypes of hepatitis B virus were determined by Nested polymerase chain reaction(nt PCR).The levels of serum HBVDNA replication,ALT levels and HBV serologic markers were detected pre or post -treatment 24 weeks, 48 weeks.Observed the relationship between the HBV genotypes/sub -genotypes and the antiviral efficacy of adefovir dipivoxil or entecavir treatment.Results After 24 weeks of adefovir dipivoxil therapy,ALT normalization rate of subgenotype Ba and subgenotype C2 was 66.7% vs 66.2%(χ2 =0.74,P >0.05),HBeAg negative conversion rate of two subgenotypes was 27.3% vs 23.1%(χ2 =0.10,P >0.05),HBVDNA negative conversion rate of two sub genotype Ba and subgenotype C2 was 33.3% vs 30.9%(χ2 =0.03,P >0.05),respectively.After treatment for 48 weeks,ALT normalization rates of subgenotype Ba and subgenotype C2 were 83.3% vs 82.4%(χ2 =0.01,P >0.05),HBeAg negative conversion rates of two subgenotypes were 45.5% vs 34.4%(χ2 =0.49,P >0.05 ),HBVDNA negative conversion rates of two subgenotypes were 58.3% vs 48.5%(χ2 =0.39,P >0.05).After 24 weeks of entecavir therapy, ALT normalization rates of subgenotype Ba and subgenotype C2 were 71.4% vs 69.6%(χ2 =0.02,P >0.05 ), HBeAg negative conversion rates of two subgenotypes were 33.3% vs 30.8%(χ2 =0.03,P >0.05 ),HBVDNA negative conversion rates of two subgenotypes were 42.9% vs 39.3%(χ2 =0.06,P >0.05 ),respectively.After treatment for 48 weeks,ALT normali zation rates of subgenotype Ba and subgenotype C2 were 85.7% vs 83.9%(χ2 =0.03,P >0.05),HBeAg negative conversion rates of two subgenotypes were 50.0% vs 44.9%(χ2 =0.10, P >0.05),HBVDNA negative conversion rates of two subgenotypes were 71.4%vs 67.8%(χ2 =0.07,P >0.05). Conclusion The study showed that genotype C(C2)is a predominant HBV genotype among people with chronic HBV infections in Shanxi province.HBV subgenotypes Ba and C2 have no significant difference in virologic,biochemi-cal,and immunologic response to ADV or ETV.

Objective The aim of the study was to investigate the mutation of precore (PreC) region of hepatitis B virus (HBV) gene and its association with hepatocellular carcinoma.Methods A total of 99 HBV patients were divided into two groups:chronic hepatitis B (CHB,55) and hepatocellular carcinoma (HCC,44).The precore gene fragment was amplified by nested PCR and analyzed by direct DNA sequencing.To analyze the relationships between HCC occurrence and age,sex,baseline HBV DNA,HBeAg status,precore region gene mutation was analyzed in two groups by using the method of case-control.Results There was significant difference between the patients with hepatocellular carcinoma and chronic HBV infection in the baseline levels of HBV DNA.In HBV precore promote region,the common point mutations were G1896A and G1899A.Compared with patients with chronic hepatitis B,point mutation G1896A,patients with liver cancer had a higher incidence of point mutation G1896A.The negative rate of HBeAg in patients with mutation with G1896A was much higher in CHB group than HCC group,the difference was significant (P ＜ 0.05).There was no significant difference between hepatocellular carcinoma and chronic HBV infection in the mutation rate of G1899A (P ＞ 0.05).The rate of insertion or deletion mutations in HBV DNA Precore gene were lower than point mutation.Conclusion HBV PC 1896 mutation was correlated with the occurrence of hepatocellular carcinoma.