Objective To observe the effects of mesenchymal stem cells (BMSCs) on the expression of Bcl2 and Bax in atrophied muscles.Methods Immature rats (80～ 100 g) were anesthetized to collect marrow from their femurs and tibias.BMSCs were isolated from the marrow,cultured and purified using the whole bone marrow adherence method.Their right hindlimbs were immobilized fiom the thigh to the paw with the knee in extension and the ankle in plantar flexion.After the modeling,24 of the male rats were divided into a sham-operation group,a BMSC group and a phosphate buffer solution (PBS) group,each of 8.The BMSC and PBS groups were injected with either approximately 106 BMSCs or an equal volume of PBS into the belly of the soleus muscle after they had been immobilized for 48 hours,while the control group did not undergo any treatment except for the injection of PBS.All of the rats were sacrificed for analysis after 14 days.Results The BMSCs were mainly spindle cells,showing radial colony arrangement.They grew vigorously and could passage in a continuous and stable manner over 10 passages.At the 4th passage the BMSCs were positive for CD44 (95.84％) and CD90 (96.00％),but negative for CD34 and CD45.Western blotting assay demonstrated that the expression of Bax protein as measured in grey-scale value (0.41±0.08)in the BMSC group was significantly lower than in the PBS group (0.63±0.10),but significantly higher than in the control group (0.14±0.11) on average.The expression of Bcl-2 (0.47±0.14) was also significantly higher in the BMSC group than in the PBS group (0.22-± 0.13),but significantly lower than in the control group (0.81 ± 0.06).Conclusion Bone marrow mesenchymal stem cells can upregulate the expression of anti-apoptotic Bcl-2 protein and downregulate that of the apoptotic Bax protein when injected early into the belly of a muscle in an immobilized limb.

Objective To establish immortalized human hepatocyte lines for studies of bioartificial liver,hepatocyte transplantation,and drug metabolism in vitro.Methods Primary human hepatocytes were isolated by 4-step perfusion technique with collagenase and transfected with recombinant retrovirus containing Simian virus 40 large T antigen(SV40 LT).Subsequently,immortalized human hepatocytes were evaluated by analysis of gene expression and functional characteristics in vitro.Results Two immortalized human hepatocyte lines,HepLi2 and HepLi3,were obtained after primary human hepatocytes being infected by SV40 LT containing recombinant retrovirus for 3-4 weeks.The immortalized human hepatocytes showed classical appearance of hepatocyte observed by phase contrast microscope.The protein expression of SV40 LT in HepLi2 and HepLi3 cells were detected by Western blotting.The mRNA expressions of albumin(Alb),glutathione S-transferase(GST-p),human blood coagulation factor X(HBCF-X)and β-actin in HepLi2 and HepLi3 cells were detected by reverse transcription polymerase chain reaction(RT-PCR),and the mRNA expressions of cytochrome(CY)450 subtypes(CYP3A5,CYP2E1,CYP2C8-19 and CYP3A4)in HepLi2 and HepLi3 cells were also observed by RT-PCR.Levels of alanine transaminase (ALT),lactate dehydrogenase(LDH)and Alb were detected in the supernatant of immortalized human hepatoeyte culture.Conclusions The immortalized human hepatocyte lines have the biological characteristics of primary human hepatocytes and have the CYP450 functions of hepatocytes,which may be heIDful for the studies of bioartificial liver,heoatocvte transplantation and drug metabolism in vitro.

【Objective】 To explore the effects of acute normovolemic hemodilution (ANH) combined with intraoperative recycled autotransfusion on blood transfusion volume and coagulation function in elderly patients undergoing cardiac surgery. 【Methods】 A total of 94 elderly patients undergoing cardiac surgery in the hospital were enrolled and randomly divided into observation group (n=47) and control group (n=47) between March 2020 and March 2022. The observation group was given ANH combined with intraoperative recycled autotransfusion, while control group was given routine allogeneic transfusion. The blood transfusion volume, oxygenation status, immune function, inflammatory indexes and adverse reactions in the two groups were compared. 【Results】 The banked blood transfusion volume was less in observation group than control group [(1.73±0.43) U vs (5.71±1.71) U, P<0.05]. At 6 h after surgery, blood oxygen saturation (SvO2) level was higher [(74.59±7.20) % vs (67.22±6.19) %], while oxygen uptake rate (ERO2) level was lower[(0.29±0.06) % vs (0.34±0.05) %] in observation group than control group (P<0.05). At day 1 after surgery, levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺ and natural killer cells (NK) were higher [(65.11±5.14) %, (46.93±5.17) %, (1.86±0.30), (8.35±1.23) % vs (57.45±7.24) %, (43.58±4.85) %, (1.47±0.36)%, (7.34±1.38) %], while CD8⁺ was lower [(25.17±4.01) % vs (30.39±5.06) %] in observation group than control group (P<0.05). At day 1 after surgery, levels of serum interleukin 6/8 (IL-6/8) and tumor necrosis factor-α (TNF-α) were lower in observation group than control group [(104.51±12.55) ng/L vs (125.81±14.96) ng/L, (351.42±52.86) ng/L vs (394.27±55.78) ng/L, (254.93±49.94) ng/L vs (323.60±52.63) ng/L, P<0.05]. The incidence of adverse reactions was lower in observation group than control group (4.26% vs 17.02%), P<0.05. 【Conclusion】 Recycled autotransfusion can reduce allogeneic transfusion volume in elderly patients undergoing cardiac surgery. There is no difference in the effects on blood routine or coagulation function between recycled autotransfusion and allogeneic transfusion. Compared with allogeneic transfusion, recycled autotransfusion can significantly improve oxygenation status, relieve immunosuppression and inflammation response, and reduce the risk of adverse reactions.

Objective:To analyze the clinical features and risk factors of invasive pulmonary aspergillosis (IPA) in patients with chronic obstructive pulmonary disease (COPD).Methods:Ninety COPD patients with IPA admitted in the First Affiliated Hospital, Zhejiang University School of Medicine and Zhejiang Provincial People’s Hospital from January 2015 to September 2019 were enrolled, and 180 COPD patients without IPA admitted in the same period were selected as control group. The clinical data of the patients in both groups were analyzed retrospectively. Chi square test was used to compare the imaging characteristics of patients in two groups, and multivariate conditional Logistic regression analysis was used to explore the risk factors of IPA in COPD patients. Results:Among 90 cases of COPD with IPA, the culture of lower respiratory tract samples identified Aspergillus fumigatus in 78 cases, Aspergillus flavus in 6 cases, Aspergillus fumigatus with Aspergillus flavus in 1 case, Aspergillu sterrus in 1 case, and Aspergillus nigerwere in 1 case; 1 case of Aspergillus mycelium was found by sputum exfoliation cytology and 2 cases were positive for serum galactomannan. Chest CT images showed patchy infiltrating shadow (87.8%), pleural effusion (36.7%), nodules (33.3%), cavity (18.9%), consolidation shadow (17.8%), halo sign (14.4%) and air crescent sign (2.2%). The incidence of patchy infiltrating shadow, consolidation shadow, halo sign and cavity were higher in COPD patients with IPA compared to control group ( P<0.05 or P<0.01). Multivariate conditional Logistic regression analysis showed that prolonged hospital stay ( OR=1.183, 95% CI 1.047-1.336), combination of two antibiotics ( OR=5.391, 95% CI 1.241-23.421), duration of antibiotic treatment ≥14 days ( OR=5.275, 95% CI 1.586-17.541), cumulative use of antibiotics ( OR=2.270, 95% CI 1.406-3.664) were the risk factors of COPD with IPA ( P<0.05 or P<0.01). Conclusion:The risk factors of IPA in COPD patients include long duration of hospital stay, combination of two kinds of antibiotics, more than 14 days of antibiotic treatment, and more varieties of antibiotics. If the above risk factors exist in patients with COPD, etiology and serology examination and dynamic monitoring of chest CT scan should be performed for early diagnosis and improved prognosis.

Accumulating evidence has confirmed the links between transfer RNA (tRNA) modifications and tumor progression. The present study is the first to explore the role of tRNA methyltransferase 5 (TRMT5), which catalyzes the m1G37 modification of mitochondrial tRNAs in hepatocellular carcinoma (HCC) progression. Here, based on bioinformatics and clinical analyses, we identified that TRMT5 expression was upregulated in HCC, which correlated with poor prognosis. Silencing TRMT5 attenuated HCC proliferation and metastasis both in vivo and in vitro, which may be partially explained by declined extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Mechanistically, we discovered that knockdown of TRMT5 inactivated the hypoxia-inducible factor-1 (HIF-1) signaling pathway by preventing HIF-1α stability through the enhancement of cellular oxygen content. Moreover, our data indicated that inhibition of TRMT5 sensitized HCC to doxorubicin by adjusting HIF-‍1α. In conclusion, our study revealed that targeting TRMT5 could inhibit HCC progression and increase the susceptibility of tumor cells to chemotherapy drugs. Thus, TRMT5 might be a carcinogenesis candidate gene that could serve as a potential target for HCC therapy.
Humans ; Carcinoma, Hepatocellular/pathology* ; Cell Hypoxia ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Liver Neoplasms/pathology* ; Signal Transduction/genetics* ; tRNA Methyltransferases/metabolism*