Potential biomarkers for the prognosis and treatment of HCC immunotherapy.

OBJECTIVE: The liver is a unique organ containing large populations of immune cells. Immunotherapy for liver cancer is a promising yet particularly challenging method. Therefore, it harbors great significance for the identification of immune-related subtypes and the potential therapeutic targets for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Firstly, we classified the HCC samples downloaded from the dataset of Cancer Genome Atlas (TCGA) into two clusters based on the immune cell infiltration. Thereafter, we identified the significant module and regulatory factors using the weighted gene co-expression network analysis (WGCNA). The immune competence of the regulatory factors was delineated through the ESTIMATE algorithm, the analysis of the tumor microenvironment, and pan-cancer analysis. In the single-cell RNA sequencing analysis, we further explored the immune competence of regulatory factors. We also collected the potential drugs targeting the regulatory factors. In addition, we constructed lncRNA-miRNA-mRNA interaction regulatory networks. Finally, western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were conducted to verify the protein expression of regulatory genes in HCC cell lines and tissues. RESULTS: According to the immune cell infiltration, two immune-related subtypes-cluster 1 and cluster 2-were found. Patients in cluster 2 had a more significant immune infiltration than in cluster 1. Afterward, six significant regulatory genes were identified through WGCNA, and the expression in cluster 2 was high in cluster 1. We performed a comprehensive analysis to clarify the immune signature. The results showed that the six genes had significant immunological competence. Moreover, the expression of the six genes was similar to the subtypes' classification. In the analysis of the prognosis value, patients in cluster 2 had a better prognosis. In addition, the lncRNA in the lncRNA-miRNA-mRNA interaction regulatory networks was located in the nucleus and cytoplasm. In the single-cell RNA sequencing analysis, the six genes were related to the immune cell. We also identified potential drugs for CD6 and CLEC12A, which may provide potential therapeutic drugs. Finally, the regulatory genes were verified in the western blot and quantitative real-time polymerase chain reaction. CONCLUSIONS: The classification into two clusters based on the immune cell infiltration may provide a promising prospect for HCC through immunotherapy. The six regulatory genes may be potential therapeutic targets in the treatment of HCC.

[Comparative study of decentration, tilt and visual quality after implantation of aspherical intraocular lenses].

Objective: To compare the difference of decentration and tilt among 4 kinds of aspherical intraocular lenses (IOLs), and to analyze their objective visual quality. Methods: Prospective non-randomized controlled clinical trial. Age-related cataract patients who planned to undergo phacoemulsification and IOL implantation in the Affiliated Hospital of North Sichuan Medical College from April to June in 2020 were divided into ZCB00 group, SN60WF group, A1-UV group, and AO group according to IOL types. Thirty right eyes and thirty left eyes were selected in each group. Before operation and at 1 day, 1 week and 1 month postoperatively, decentration and tilt values were measured by a new swept-source anterior-segment optical coherence tomography device (CASIA2), and wavefront aberrations, objective scatter index (OSI), modulation transfer function cut off frequency (MTF cut off) and Strehl ratio (SR) were also examined. Values of decentration, tilt and visual quality compared among 4 groups were took from the right eye. One-way analysis of variance was used for inter-group comparison, and repeated measurement one-way analysis of variance was used for intra-group comparison. Data between right eyes and left eyes from all the individuals were compared by independent sample t-test. Results: A total of 181 patients (240 eyes) were enrolled, and 169 patients (224 eyes) completed the follow-up (114 right eyes and 110 left eyes). There were 77 males and 92 females, with an age of (69±9) years. There was no significant difference in gender, age, axial length, decentration and tilt of crystalline lens and IOL power among 4 groups (all P>0.05). At 1 day, 1 week and 1 month postoperatively, there was significant difference in decentration value among 4 groups (F=7.11, 6.12, 4.66; all P<0.05). For further pairwise comparison, the decentration value of SN60WF group was higher than that of the other 3 groups at 1 day and 1 week postoperatively, and the decentration value of SN60WF group was (0.259±0.101) mm at 1 month postoperatively, which was higher than that of ZCB00 group (0.177±0.099) mm and AO group (0.163±0.122) mm, and the differences were statistically significant (using SNK-q test, both P<0.05). The IOL tilt value in the ZCB00 group, SN60WF group, A1-UV group, and AO group at 1 month postoperatively were (4.806±1.129)°, (5.080±1.309)°, (4.586±1.338)°, (5.112±1.406)°, respectively. No significant difference in tilt value among 4 groups was found at any time after surgery (all P>0.05). In each group, there was no significant difference in decentration and tilt value at different postoperative time points (all P>0.05). At 1 month postoperatively, there was no significant difference in decentration and tilt at horizontal and vertical directions respectively among 4 groups (all P>0.05), and in each group, there was no significant difference in decentration and tilt value between right eyes and left eyes (all P>0.05), and IOLs tended to tilt towards the inferonasal or inferotemporal direction in both eyes. With 4-mm and 6-mm pupil diameter, there was significant difference in internal (F=131.32, 85.17) and ocular (F=46.64, 47.55) spherical aberration among 4 groups (all P<0.01). For further pairwise comparison, the spherical aberration of AO group was higher than that of the other 3 groups, and the difference was statistically significant (using SNK-q test, all P<0.05). There was no significant difference in OSI, MTF cut off and SR among 4 groups (all P>0.05). Conclusions: The four types of IOLs show decentration and tilt in varying degrees after implantation in the capsular bag, but this difference do not lead to clinical significance. Human eyes have tolerance to mild decentration and tilt of aspheric IOLs, showing good visual quality.

Simvastatin relieves myocardial ischemia/reperfusion injury in rats through hedgehog signaling pathway.

OBJECTIVE: The aim of this study was to explore the effect of simvastatin (Sim) on myocardial ischemia/reperfusion (I/R) injury in rats and to elucidate the possible underlying mechanism. Our findings might help to provide a certain reference for the clinical prevention and treatment of myocardial I/R injury. MATERIALS AND METHODS: A total of 60 male Sprague-Dawley (SD) rats were randomized into three groups using a random number table, including: Sham group (n=20), I/R group (n=20) and I/R + Sim group (n=20). The I/R injury model was successfully established in rats via ligation of the left anterior descending coronary artery (LAD), followed by reperfusion. Before operation, the rats in I/R + Sim group were administered with Sim at 10 mg/kg/d through oral gavage for 7 d. Cardiac ejection fraction (EF) (%) and fractional shortening (FS) (%) of rats in each group were detected using echocardiography. 2,3,5-triphenyltetrazolium chloride (TTC) staining was performed to measure the myocardial infarction (MI) area in each group. Collagen deposition in myocardial tissues of rats in each group was detected by Masson's trichrome staining. The apoptosis level of myocardial cells and fibroblasts in myocardial tissues of rats in each group were evaluated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. The level of reactive oxygen species (ROS) in myocardial tissues of rats in each group was determined using fluorescent probes. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was conducted to measure the expression levels of pro-inflammatory cytokines interleukin (IL)-1ß and monocyte chemoattractant protein-1 (MCP-1) in myocardial tissues of rats in each group. Furthermore, the effects of Sim on the hedgehog signaling pathway-associated proteins were detected using Western blotting. RESULTS: Sim significantly alleviated I/R-induced cardiac dysfunction in rats and increased EF (%) and FS (%) (p<0.05). Meanwhile, it also evidently mitigated the MI caused by I/R and reduced the infarction area (p<0.05). According to the Masson's trichrome staining results, I/R + Sim group exhibited remarkably declined myocardial interstitial collagen deposition compared with I/R group (p<0.05). ROS-sensitive fluorescent staining showed that Sim notably reversed the increase of ROS expression and the decrease of myocardial oxidative stress induced by I/R (p<0.05). Finally, Western blotting results revealed that Sim dramatically restrained the protein expressions of sonic hedgehog (SHH), patched 1 (PTC1) and glioma-associated oncogene homolog 1 (GLI1) (p<0.05). CONCLUSIONS: Sim can significantly relieve myocardial I/R injury in rats. The possible underlying mechanism may be related to its inhibition on the hedgehog signaling pathway.

Alterations of immune functions induced by 12C6+ ion irradiation in mice.

PURPOSE: To estimate the biological risks to the immune system of the type of space radiation, 12C6+, encountered by cosmonauts during long-term travel in space. MATERIALS AND METHODS: The Kun-Ming strain mice were whole-body irradiated by 12C6+ ion with 0, 0.01, 0.05, 0.075, 0.2, 0.3, 0.5, 0.75, 1 or 2 Gy, at a dose rate of 1 Gy/min. At 35 days after irradiation, the thymus and spleen weights were measured, the natural killer (NK) cells activity of spleen was determined by 3-(4, 5-dimethylthiazol-2-yl)- 2, 5-diphenyl tetrazolium bromide (MTT), and the interferon-gamma (IFN-gamma) levels in serum and thymus were detected with enzyme-linked immunosorbent assays (ELISA). RESULTS: The results showed that the thymus weight, IFN-gamma levels in serum and the activity of splenic NK-cells had significantly increased at a dose of 0.05 Gy. With further dose increase, the weight of spleen continued to increase but the weight of thymus, IFN-gamma level and NK-cells activity declined. CONCLUSIONS: These results suggest that the dose of 0.05 Gy irradiation has a stimulatory effect on mouse immunity; this effect declined with increasing dose.

Binding of the maize cytosolic Hsp70 to calmodulin, and identification of calmodulin-binding site in Hsp70.

Using a gel-overlay technique of biotinylated calmodulin (CaM), we showed that maize cytosolic Hsp70 protein could bind to CaM in the presence of 1 mM CaCl2. The purified maize cytosolic Hsp70 inhibited the activity of CaM-dependent NADK in a concentration-dependent manner. A synthetic peptide, which possesses the 21 amino acid sequence, PRALRRLRTACERAKRTLSST, at positions 261-281 in maize cytosolic Hsp70, could associate with CaM in the presence of 1 mM calcium. The synthetic peptide inhibited CaM-dependent NADK activity and PDE activity. This indicates that the 21-amino acid sequence at positions 261-281 is the CaM-binding site. The binding of CaM to Hsp70 inhibited the ATPase activity of Hsp70. The possible regulator function of Hsp70 in cell signaling events in response to heat stress is discussed.

Orphanin FQ/nociceptin modulates glutamate- and kainic acid-induced currents in acutely isolated rat spinal dorsal horn neurons.

The heptadecapeptide orphanin FQ (OFQ), also known as nociceptin (NOC), is a newly discovered endogenous ligand for the opioid-like G-protein coupled receptor, ORL1. In the present study, the effects of OFQ/NOC on glutamate (Glu), kainic acid (KA) and quisqualic acid (QA) induced currents were examined in isolated rat spinal dorsal horn neurons of young rats using whole-cell patch-clamp techniques. Glu, KA and QA elicited rapid inward currents in 90%, 69%, 83% of tested neurons. OFQ/NOC(0.03 approximately 300 nM) failed to induce any changes of membrane currents, but modulated Glu-, KA- and QA-elicited currents. OFQ/NOC inhibited and potentiated Glu-induced currents in 40.6% and 27.3% of examined cells (n=106) respectively. In about one third examined neurons, OFQ/NOC had no detectable effects on Glu responses. OFQ/NOC also inhibited and enhanced KA- and QA-induced currents (inhibition: KA, 67.1%, n=76; QA, 50%, n=36. Potentiation: KA, 23.7%, n=76; QA, 16.7%, n=36). In about 10% of tested cells, Glu-induced currents were potentiated after the application of OFQ/NOC, and lasted for 20 approximately 30 min. The inhibitory effects of OFQ/NOC on KA and QA responses were naloxone-insensitive. The C-terminal fragment OFQ(8-17) presented same effects on EAA-induced responses. Taken together, OFQ/NOC primarily inhibited Glu-, KA- and QA-induced currents in isolated rat spinal dorsal horn neurons via non-opioid mechanism, which might contribute to nociceptive transmission in the spinal level.

Antimicrobial susceptibilities and beta-lactamase production of Hong Kong isolates of gastroenteric salmonellae and Salmonella typhi.

We examined the in-vitro antibiotic susceptibility of 760 gastroenteric salmonellae and 36 strains of Salmonella typhi isolated in Hong Kong between 1985 and 1988. S. typhi remained susceptible to all the antibiotics tested except for one isolate resistant to chloramphenicol, another to kanamycin and co-trimoxazole, and a third to nalidixic acid. In contrast, resistance and multiple resistance has increased significantly in gastroenteric salmonellae over the last ten years. Seventeen percent were resistant to ampicillin, 61% to tetracycline, 23% to chloramphenicol and 8% to gentamicin. Many ampicillin-resistant strains remained resistant to ampicillin even in the presence of sulbactam (69%) or clavulanic acid (25%). More than 50% of isolates were resistant to two or more antibiotics and one isolate was resistant to eleven. Ampicillin-resistance was usually due to the production of TEM-1 or OXA-1 beta-lactamases but a few isolates produced AER-1, PSE-1 or PSE-2. Genetic determinants for these enzymes were usually borne on plasmids ranging in size from 2 to 143.7 Md but half of the OXA-1 genes were chromosomally located.

[Bacteriologic investigation of the hospital air-conditioning water].

We have isolated a pathogenic strain of LDB from the water of the hospital cooling-water storage tank for air-conditioning system. At the same time, other bacteria were also isolated with Pseudomonas and Flavobacterium being predominant, most of these organisms in the water of air-conditioning system were resistant to antibacterial drugs. A proposal to improve architectural design and to sterilize air-conditioning water was suggested.