[Effect of Liraglutide on platelet distribution width and carotid intima-media thickness in type 2 diabetic mellitus patients with obesity].

Objective: To investigate the effect of Liraglutide on platelet distribution width(PDW) and carotid intima-media thickness(cIMT) in type 2 diabetic mellitus patients with obesity. Methods: Randomized controlled trial. A total of 80 type 2 diabetes mellitus (T2DM) obese patients with unsatisfactory glucose control were prospectively enrolled in this study from the Department of Endocrinology of Yuhuangding Hospital Affiliated to Qingdao University from January to December 2021. All the participants were treated with metformin or sulfonylureas. They were randomly divided into two groups: Liraglutide treatment group (Li group, n=40) and Control group (Con group, n=40).The Li group started the treatment with Liraglutide on the basis of the original hypoglycemic agents and the Con group was treated with metformin and sulfonylurea. After 16 weeks of treatment, the changes of PDW, cIMT and body mass index (BMI) in the two groups were observed, multiple linear regression was uesd to analyze the influencing factors of cIMT variation, and the effect of liraglutide on PDW and cIMT in obese patients with type 2 diabetes was analyzed. Results: Finally, 38 patients completed the study in Li group, including 23 males and 15 females, aged 30-69(56±11) years. All 40 patients in Con group completed the study, including 18 males and 22 females, aged 39-67(59±7) years. After 16 weeks of treatment, the levels of PDW and cIMT in Li group were (12.8±1.6) fl and (0.85±0.08) mm, respectively, lower than those before treatment (15.0±1.6) fl and (1.14±0.10) mm (t=18.61 and 20.37, respectively, both P<0.001); The PDW and cIMT in Con group were (13.6±1.5) fl and (1.05±0.10) mm, respectively, lower than those before treatment (15.0±1.5) fl and (1.13±0.13) mm (t=17.42 and 9.65, respectively, both P<0.001). The levels of fasting plasma glucose (FPG) and total cholesterol (TC) in both groups were lower than those before treatment(all P<0.001). After the treatment, the levels of PDW, cIMT, FPG and TC in Li group were lower than those in Con group (all P<0.05). The changes of PDW and cIMT before and after the treatment in Li group were (2.2±0.7) fl and (0.30±0.09) mm, respectively, higher than those in the Con group [(1.4±0.5) fl and (0.09±0.06) mm], with a statistically significant difference (both P<0.001). The changes of FPG and TC in Li group were significantly higher than those in Con group (all P<0.05). Multiple linear regression analysis showed that liraglutide, the changes of TC and systolic blood pressure (SBP) were the influencing factors for the changes of cIMT [ß (95%CI) were 0.20 (0.17-0.23), 0.03 (0.01-0.06), 0.01 (0.00-0.01), respectively, all P<0.05] Conclusion: Liraglutide treatment could reduce PDW and cIMT, thus contributing to cardiovascular benefits.

Silence of ATAD2 inhibits proliferation of colorectal carcinoma via the Rb-E2F1 signaling.

OBJECTIVE: This study aims to clarify the potential function of ATAD2 (ATPase family, AAA domain containing 2) in regulating proliferative and apoptotic abilities of colorectal carcinoma (CRC). PATIENTS AND METHODS: ATAD2 levels in CRC specimens and cell lines were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Overall survival in CRC patients with high or low level of ATAD2 was assessed by Kaplan-Meier method. The correlation between ATAD2 level and clinical characteristics of CRC patients was analyzed by χ2 test. Univariable and multivariable Cox regression models were generated to illustrate potential risk factors for the overall survival of CRC. After knockdown of ATAD2 in SW620 cells, relative levels of Cyclin D1, ppRb, pRb, E2F1, Cyclin E and cleaved Caspase 3 were detected by Western blot. Regulatory effects of ATAD2 on viability, clonality, cell cycle distribution, and apoptosis in SW620 and HCT15 cells were examined by a series of functional experiments. RESULTS: Upregulated ATAD2 in CRC was correlated to tumor size, tumor node metastasis (TNM) staging, and histological classification of CRC. High level of ATAD2 predicted poor prognosis in CRC patients. Cox regression test suggested that ATAD2 level, tumor size, TNM staging and histological classification were independent factors influencing overall survival in CRC. Knockdown of ATAD2 reduced viability and clonality in SW620 and HCT15 cells. In addition, cell cycle was arrested in G1 phase and apoptosis was stimulated in CRC cells with ATAD2 knockdown. In SW620 cells transfected with ATAD2 shRNA, protein levels of Cyclin D1, ppRb, E2F1 and Cyclin E were downregulated, and cleaved Caspase 3 was upregulated. CONCLUSIONS: ATAD2 is upregulated in CRC tissues and correlated to poor prognosis of CRC patients. It exerts an anti-proliferation role in CRC by arresting cell cycle in G1/S phase and triggering apoptosis via the Rb-E2F1 signaling.

Establishment of patient-derived tumor xenograft (PDTX) models using samples from CT-guided percutaneous biopsy.

This study aimed to investigate the feasibility of the establishment of a human cancer xenograft model using samples from computed tomography (CT)-guided percutaneous biopsy. Fresh tumor tissues obtained from 10 cancer patients by CT-guided percutaneous biopsy were subcutaneously inoculated into NOD-Prkdcem26Il2rgem26Nju (NCG) mice to establish human patient-derived tumor xenograft (PDTX) models. The formation of first and second generation xenografts was observed, and tumor volume was recorded over time. Tumor tissue consistency between the PDTX model and primary tumors in patients was compared using H&E staining and immunohistochemistry. Pharmacodynamic tests of clinically used chemotherapeutic drugs were conducted on second generation xenografts, and their effects on tumor growth and body weight were observed. CT-guided percutaneous biopsy samples were successfully collected from 10 patients with advanced cancers. The PDTX model was established in mice using tumor samples obtained from 4 cancer patients, including one small cell carcinoma sample, two adenocarcinoma samples, and one squamous cell carcinoma sample. The success rate was 40%. The obtained PDTX model maintained a degree of differentiation, and morphological and structural characteristics were similar to primary tumors. The pharmacodynamic test of chemotherapeutic drugs in the PDTX model revealed a therapeutic effect on tumor growth, as expected. CT-guided percutaneous biopsy samples can be effectively used to establish a PDTX model, and test these chemotherapy regimens.

Correlation between liver cancer occurrence and gene expression profiles in rat liver tissue.

Liver cancer (LC) is generally characterized by malignant cell proliferation and growth; it normally develops in stages that progress from non-specific injury of the liver to liver fibrosis, liver cirrhosis, dysplasia nodules, and liver carcinoma. We used a rat model of diethylnitrosamine (DENA)-induced LC; a Rat Genome 230 2.0 Array was used to detect gene expression profile of liver tissues from male rats 5, 8, 12, 16, and 18 weeks following the beginning of DENA-induced LC. We found 909 known genes, including 637 up-regulated, 270 down-regulated, and two up/down-regulated genes, that were significantly changed in expression. Among them, 108 genes were expressed at the 5th, 213 at the 8th, 516 at the 12th, 698 at the 16th, and 506 at the 18th week of DENA-induced LC. Methods in bioinformatics and systems biology were applied to explore the correlation between the gene expression profile of rat liver tissue and liver cancer occurrence at the transcriptional level; 23 physiological activities were found to be associated with LC. Among these, eight physiological activities, including stimulus response, inflammation and immune response, oxidative reduction, cell proliferation, differentiation, migration, adhesion, and angiogenesis were increased, implying that they could play important roles in the occurrence and development of LC. In addition, carbohydrate, lipid, and organic acid metabolism were decreased, suggesting that liver injury induced by a carcinogenic agent has a negative effect on the metabolism of fundamental substances.

CD4+ T cells are required for HSP65 expression in host macrophages and for protection of mice infected with Plasmodium yoelii.

We have reported that macrophages expressing heat-shock protein 65 play an essential role in protection of mice infected with Plasmodium yoelii. In this study, we investigated the function and expression mechanism of HSP65 in macrophages of mice infected with P. yoelii. C57BL/6 (B6) mice are susceptible to infection with the lethal (L) strain but resistant to infection with the non-lethal (NL) strain of P. yoelii. The percentage of apoptotic macrophages in mice infected with the L strain was higher than that in mice infected with the NL strain. However, the percentage was low in L strain infected mice if they acquired resistance to the infection by primary infection with the NL strain. That apoptosis was reversely correlated with HSP65 expression in splenic macrophages from mice infected with P. yoelii suggests HSP65 may contribute to protective immunity by preventing apoptosis of macrophages in malarial infection. Cell depletion/transfer experiments showed that CD4+ T cells, but not CD8+ T cells, gammadelta T cells, NK cells or NK T cells, were required for HSP65 expression in macrophages as well as for protection of mice infected with P. yoelii. In conclusion, HSP65 may play a role in preventing apoptosis of macrophages in mice infected with P. yoelii. CD4+ T cells are required for HSP65 expression and for protective immunity against P. yoelii infection.

Antibodies specific for heat shock proteins in human and murine malaria.

Heat shock proteins (HSPs) are immunodominant antigens recognized by the host immune system in various infectious diseases. We analyzed HSP-specific antibodies, including immunoglobulin G (IgG), IgM and IgA, in sera from malaria patients in Thailand by using an enzyme-linked immunosorbent assay. All of the antibodies to HSP90 were remarkably increased in the patients compared with those in controls, while only IgM to HSP70 or IgA to HSP65 was significantly elevated. Further experiments showed that anti-HSP IgG was significantly increased in C57BL/6 mice infected with a non-lethal strain of Plasmodium yoelii, with anti-HSP90 IgG being the most elevated. These results suggest that the antigenic potential of HSP90 is higher than those of HSP70 and HSP65 in malaria infection.

Macrophages expressing heat-shock protein 65 play an essential role in protection of mice infected with Plasmodium yoelii.

C57BL/6 (B6) mice are resistant to infection with the non-lethal (NL) strain of Plasmodium yoelii 17X, while being susceptible to that with the lethal (L) strain. The 65 000 MW heat-shock protein (hsp 65) was strongly expressed in splenic adherent cells of B6 mice 10 days after infection with the NL strain of P. yoelii but only slightly in those from mice infected with the L strain. Mice which had survived infection with the NL strain were resistant to challenge with the L strain and hsp 65 was strongly expressed in splenic adherent cells of these mice. Severe combined immunodeficient mice and nude mice were susceptible to malaria infection even with the NL strain and did not express hsp 65 after infection, suggesting that T cells are required for the expression of hsp 65 as well as for protective immunity. B6 mice treated intraperitoneally with carrageenan, which impairs the macrophage function, became susceptible to NL strain infection, indicating that macrophages play an important role as the final effectors in protective immunity. These results demonstrate that the hsp 65 expressed by macrophages correlates closely with protection against P. yoelii infection.

Serotyping and subgrouping of some rotavirus strains in China.

This article reports for the first time in China the results of serotyping and subgrouping of some rotavirus strains obtained from faecal fluids of cases with infant diarrhoea. Infant diarrhoea rotavirus serotype 1 (Wa strain)-specific and serotype 2 (KUN strain)-specific hyperimmune sera, and infant diarrhoea rotavirus serotype 3-specific, rotavirus subgroup I-specific and subgroup II-specific monoclonal antibodies were used in serotyping and subgrouping by means of enzyme-linked immunosorbent assay of 14 rotavirus strains which had been identified by electron microscopy. Results showed that three out of four Beijing strains in 1982 belonged to serotype 2 and subgroup I rotavirus, one belonged to serotype 3 and subgroup II rotavirus; one out of nine Beijing strains in 1984 belonged to serotype 1 and subgroup II rotavirus, seven belonged to serotype 2 and subgroup I rotavirus, one belonged to serotype 3 and subgroup II rotavirus; and one Kunming strain in 1984 belonged to serotype 3 and subgroup II rotavirus.

Antibody against adult diarrhoea rotavirus among healthy adult population in China.

This paper deals with a method of using unknown serum as the first antibody to coat wells of the microtiter plate directly in application of the sandwich ELISA method with double antibodies for the assay of the antibody against adult diarrhoea rotavirus. This method was used for assay of the antibody against adult rotavirus in 1,380 sera of healthy adults obtained from some provinces and cities of China, of which 141 showed a positive result with a total positive rate of 10.2%. The infection rate varied in different regions. In order to confirm the accuracy of these results a blocking test was carried out on 25 positive specimens, and the results revealed that the P/N ratios after blocking were all lower than those before blocking. Seventeen of these 25 positive serum specimens were obtained from rural areas of Qian'an County, Hebei Province where epidemic adult diarrhoea had occurred two years ago. The antibody against adult diarrhoea rotavirus was not detected in 50 adult sera from Xizang Autonomous Region. Only one out of 50 adult sera from Hainan Island was positive for the antibody.