Objective To explore the efficacy and safety of recombinant human anti-severe acute respiratory syn-drome coronavirus 2(anti-SARS-CoV-2)monoclonal antibody injection(F61 injection)in the treatment of patients with coronavirus disease 2019(COVID-19)combined with renal damage.Methods Patients with COVID-19 and renal damage who visited the PLA General Hospital from January to February 2023 were selected.Subjects were randomly divided into two groups.Control group was treated with conventional anti-COVID-19 therapy,while trial group was treated with conventional anti-COVID-19 therapy combined with F61 injection.A 15-day follow-up was conducted after drug administration.Clinical symptoms,laboratory tests,electrocardiogram,and chest CT of pa-tients were performed to analyze the efficacy and safety of F61 injection.Results Twelve subjects(7 in trial group and 5 in control group)were included in study.Neither group had any clinical progression or death cases.The ave-rage time for negative conversion of nucleic acid of SARS-CoV-2 in control group and trial group were 3.2 days and 1.57 days(P=0.046),respectively.The scores of COVID-19 related target symptom in the trial group on the 3rd and 5th day after medication were both lower than those of the control group(both P＜0.05).According to the clinical staging and World Health Organization 10-point graded disease progression scale,both groups of subjects improved but didn't show statistical differences(P＞0.05).For safety,trial group didn't present any infusion-re-lated adverse event.Subjects in both groups demonstrated varying degrees of elevated blood glucose,elevated urine glucose,elevated urobilinogen,positive urine casts,and cardiac arrhythmia,but the differences were not statistica-lly significant(all P＞0.05).Conclusion F61 injection has initially demonstrated safety and clinical benefit in trea-ting patients with COVID-19 combined with renal damage.As the domestically produced drug,it has good clinical accessibility and may provide more options for clinical practice.

Magnetic separation(MS)is frequently used to enrich specific targets from biological sam-ples,which is commonly performed using antibodies coupled immunomagnetic beads(IMBs).However,due to the high cost of antibodies and difficulties in the process of releasing captured targets,IMBs have limitations for large-scale enrichment of targeted bioproducts.The specificities and affinities of aptamers towards their targets are in line with antibodies,but with characteristic of significantly lower costs of prep-aration,simpler structures,and higher chemical stability.In this study,taking advantages of the distinct adsorption features of graphene for single-stranded and double-stranded nucleic acids,we designed a novo system as bioseparation method that could display aptamers on graphene surfaces to enrich targets and then manipulate their release.This system mainly includes two units.Take CD63 aptamer as an exam-ple:double-stranded oligonucleotides scaffold attached to the graphene surface for CD63 aptamer exhibi-tion;and a single-stranded oligonucleotide complementary to the scaffold bipod sequence to desorb the captured targets from graphene.In this study,firstly,using graphene oxide(GO)paper as a graphene surface,we fluorescently labeled scaffolds or CD63 proteins and compared the fluorescence intensity difference on the GO paper before and after scaffold or CD63 protein release.Subsequently,a novo mate-rial,amino-modified graphene-shelled iron-nitrogen magnetic beads were introduced to carry the scaffold,and used to capture CD63 proteins from cell lysates.The results indicate that this scaffold can display the aptamer on the graphene surface for CD63 protein capture and controlled release.Using selected modified graphene magnetic beads carried with the scaffold,we achieved the capture of CD63 proteins from cell ly-sates.This system is expected to enrich various proteins such as CD63,or capture exosomes by hooking membrane proteins.

With the development of high technology, people spend more and more time on screens. Most screens contain a lot of blue light. As one of the important components of visible light, blue light has high energy. It may lead to many ocular diseases, such as myopia, cataract, dry eye, glaucoma and keratitis when eyes exposure to blue light for a long time. At present, the harm of blue light and how to prevent blue light have become a hot topic. Its mechanism mainly includes increasing the photosensitivity of lipofuscin, destroying the mitochondria, lysosome, lens and tear film. According to the mechanism of blue light damage, physical protective measures can be adopted. And the antioxidant, free radical scavengers, anti-inflammatory drugs and gene therapy can be used to protect eye tissue. This paper mainly summarizes the harmful mechanism of blue light to eye and the corresponding prevention and treatment measures.

This study explored the molecular mechanism of acteoside against hepatoma 22(H22) tumor in mice through c-Jun N-terminal kinase(JNK) signaling pathway. H22 cells were subcutaneously inoculated in 50 male BALB/c mice, and then the model mice were classified into model group, low-dose, medium-dose, and high-dose acteoside groups, and cisplatin group. The administration lasted 2 weeks for each group(5 consecutive days/week). The general conditions of mice in each group, such as mental status, diet intake, water intake, activity, and fur were observed. The body weight, tumor volume, tumor weight, and tumor-inhibiting rate were compared before and after administration. Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE) staining, and the expression of phosphorylated(p)-JNK, JNK, B-cell lymphoma-2(Bcl-2), Beclin-1, and light chain 3(LC3) in each tissue was detected by immunohistochemistry and Western blot. qRT-PCR was performed to detect the mRNA expression of JNK, Bcl-2, Beclin-1, and LC3. The general conditions of mice in model and low-dose acteoside groups were poor, while the general conditions of mice in the remaining three groups were improved. The body weight of mice in medium-dose acteoside group, high-dose acteoside group, and cisplatin group was smaller than that in model group(P<0.01). The tumor volume in model group was insignificantly different from that in low-dose acteoside group, and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group. Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001). The tumor-inhibiting rates were 10.72%, 40.32%, 53.79%, and 56.44% in the low-dose, medium-dose, and high-dose acteoside groups and cisplatin group, respectively. HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups, and the necrosis was particularly obvious in the high-dose acteoside group and cisplatin group. Immunohistochemical results suggested that the expression of Beclin-1, LC3, p-JNK, and JNK was up-regulated in acteoside and cisplatin groups(P<0.05). The results of immunohistochemistry, Western blot, and qRT-PCR indicated that the expression of Bcl-2 was down-regulated in the medium-dose and high-dose acteoside groups and cisplatin group(P<0.01). Western blot showed that the expression of Beclin-1, LC3, and p-JNK was up-regulated in acteoside and cisplatin groups(P<0.01), and there was no difference in the expression of JNK among groups. qRT-PCR results showed that the levels of Beclin-1 and LC3 mRNA were up-regulated in the acteoside and cisplatin groups(P<0.05), and the level of JNK mRNA was up-regulated in medium-dose and high-dose acteoside groups and cisplatin group(P<0.001). Acteoside promotes apoptosis and autophagy of H22 cells in mice hepatoma cells by up-regulating the JNK signaling pathway, thus inhibiting tumor growth.
Male ; Animals ; Mice ; Cisplatin/pharmacology* ; Carcinoma, Hepatocellular/genetics* ; MAP Kinase Signaling System ; Beclin-1 ; Apoptosis ; Liver Neoplasms/genetics* ; Necrosis ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Cell Line, Tumor ; RNA, Messenger/metabolism* ; Autophagy

Hepatotoxicity induced by bioactive constituents in traditional Chinese medicines or herbs,such as bavachin(BV)in Fructus Psoraleae,has a prolonged latency to overt drug-induced liver injury in the clinic.Several studies have described BV-induced liver damage and underlying toxicity mechanisms,but little attention has been paid to the deciphering of organisms or cellular responses to BV at no-observed-adverse-effect level,and the underlying molecular mechanisms and specific indicators are also lacking during the asymptomatic phase,making it much harder for early recognition of hepatotoxicity.Here,we treated mice with BV for 7 days and did not detect any abnormalities in biochemical tests,but found subtle steatosis in BV-treated hepatocytes.We then profiled the gene expression of hepatocytes and non-parenchymal cells at single-cell resolution and discovered three types of hepatocyte subsets in the BV-treated liver.Among these,the hepa3 subtype suffered from a vast alteration in lipid metabolism,which was characterized by enhanced expression of apolipoproteins,carboxylesterases,and stearoyl-CoA desaturase 1(Scd1).In particular,increased Scd1 promoted monounsaturated fatty acids(MUFAs)syn-thesis and was considered to be related to BV-induced steatosis and polyunsaturated fatty acids(PUFAs)generation,which participates in the initiation of ferroptosis.Additionally,we demonstrated that mul-tiple intrinsic transcription factors,including Srebf1 and Hnf4a,and extrinsic signals from niche cells may regulate the above-mentioned molecular events in BV-treated hepatocytes.Collectively,our study deciphered the features of hepatocytes in response to BV insult,decoded the underlying molecular mechanisms,and suggested that Scd1 could be a hub molecule for the prediction of hepatotoxicity at an early stage.

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

Objective: To trace and characterize the whole genome of SARS-CoV-2 of confirmed cases in the outbreak of COVID-19 on July 31, 2021 in Henan Province. Method: Genome-wide sequencing and comparative analysis were performed on positive nucleic acid samples of SARS-CoV-2 from 167 local cases related to the epidemic on July 31, 2021, to analyze the consistency and evolution of the whole genome sequence of virus. Results: Through high-throughput sequencing, a total of 106 cases of SARS-CoV-2 whole genome sequences were obtained. The results of genome analysis showed that the whole genome sequences of 106 cases belonged to the VOC/Delta variant strain (B.1.617.2 clade), and the whole genome sequences of 106 cases were shared with the genomes of 3 imported cases from Myanmar admitted to a hospital in Zhengzhou. On the basis of 45 nucleotide sites, 1-5 nucleotide variation sites were added, and the genome sequence was highly homologous. Conclusion: Combined with the comprehensive analysis of viral genomics, transmission path simulation experiments and epidemiology, it is determined that the local new epidemic in Henan Province is caused by imported cases in the nosocomial area, and the spillover has caused localized infection in the community. At the same time, it spills over to some provincial cities and results in localized clustered epidemics.
Humans ; COVID-19 ; SARS-CoV-2/genetics* ; Genome, Viral ; Epidemics ; Phylogeny

Objective To investigate the effect of recombinant Schistosoma japonicum egg ribonuclease SjCP1412 (rSjCP1412) on proliferation, cell cycle, apoptosis and activation of human hepatic stellate cells LX-2 in vitro, and explore the underlying mechanisms. Methods The rSjCP1412 protein was expressed in Escherichia coli BL21 by prokaryotic expression, and the highly purified soluble rSjCP1412 protein was prepared by Ni NTA affinity chromatography and urea gradient refolding dialysis. Yeast RNA was digested using 12.5, 25.0, 50.0 µg rSjCP1412 proteins at 37 °C for 2, 3, 4 h, and the enzymatic products were electrophoresed on 1.5% agarose gel to observe the RNAase activity of rSjCP1412 protein. The proliferation of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was measured using CCK-8 assay, and the apoptosis of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was detected using the Annexin V-FITC/PI double staining, while the percentage of LX-2 cells at G0/G1, S and G2/M phases of cell cycle following stimulation with different doses of rSjCP1412 protein for 48 h was detected by DAPI staining. The type I collagen, type III collagen and α-smooth muscle actin (α-SMA) mRNA expression was quantified using quantitative florescent real-time PCR (qPCR) assay and Western blotting at transcriptional and translational levels in LX-2 cells following stimulation with different doses of rSjCP1412 protein for 48 h, while soluble egg antigen (SEA) served a positive control and PBS without rSjCP1412 protein as a normal control in the above experiments. The expression of collagen I, α-SMA and Smad4 protein was determined using Western blotting in LX-2 cells following stimulation with rSjCP1412 protein, transforming growth factor-β1 (TGF-β1) alone or in combination, to examine the signaling for the effect of rSjCP1412 protein on LX-2 cells. Results The rSjCP1412 protein was successfully expressed and the highly purified soluble rSjCP1412 protein was prepared, which had a RNase activity. Compared with the normal group, the survival rates of LX-2 cells significantly decreased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein and SEA for 48 h (F = 22.417 and 20.448, both P values < 0.05). The apoptotic rates of LX-2 cells significantly increased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h (F = 11.350, P < 0.05), and treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h resulted in arrest of LX-2 cells in G0/G1 phase (F = 20.710, P < 0.05). Treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h caused a significant reduction in relative expression levels of collagen I (F = 11.340, P < 0.05), collagen III (F = 456.600, P < 0.05) and α-SMA mRNA (F = 23.100, P < 0.05) in LX-2 cells, and both rSjCP1412 protein and SEA treatment caused a significant reduction in collagen I (F = 1 302.000, P < 0.05), α-SMA (F = 49.750, P < 0.05) and Smad4 protein expression (F = 52.420, P < 0.05) in LX-2 cells. In addition, rSjCP1412 protein treatment inhibited collagen I (F = 66.290, P < 0.05), α-SMA (F = 31.300, P < 0.05) and Smad4 protein expression (F = 27.010, P < 0.05) in LX-2 cells activated by TGF-β1. Conclusion rSjCP1412 protein may induce apoptosis of LX-2 cells and inhibit proliferation, cell cycle and activation of LX-2 cells through down-regulating Smad4 signaling molecules.

Objective:To investigate the clinical significance of autologous platelet separation (APS) in different time courses of cardiovascular surgery.Methods:The relevant data of 75 patients with cardiovascular surgery from September 2019 to August 2021 in Hunan Provincial Peoples′ Hospital were collected retrospectively. They were divided into two groups according to whether APS was used during the operation: group A used APS (37 cases) and group B did not use APS (38 cases). The two groups were divided into subgroups according to the length of cardiopulmonary bypass (CPB): A1 and B1 were medium and short-term groups (CPB bypass time ≤200 min), and A2 and B2 were long-term groups (CPB bypass time >200 min). Blood routine, postoperative drainage volume, postoperative blood product infusion volume and thromboelastogram at different time points were recorded and compared.Results:The postoperative drainage volume, red blood cell infusion volume and ventilator assisted time in group A were less than those in group B (all P<0.05); The postoperative drainage volume [(645.79±205.25)ml vs (886.67±360.96)ml, P=0.006], erythrocyte infusion volume [(3.24±2.53)U vs (4.77±1.97)U, P=0.016], platelet infusion volume [0.00(0.00, 0.00)U vs 1.00(0.125, 2.00)U, P=0.002] and thromboelastogram coagulation reaction time [(7.38±1.74)min vs (9.09±3.57)min, P=0.047] in group A2 were significantly better than those in group B2 (all P<0.05); There were no significant difference in the above indexes between A1 and B1 group (all P>0.05). Conclusions:APS can improve the coagulation function of patients undergoing cardiopulmonary bypass and reduce the amount of bleeding and blood products. Its protective effect is more prominent in high-risk cardiovascular surgery with long cardiopulmonary bypass and complex operation.

Objective:To evaluate the safety, feasibility and operational performance of self-developed medical disposable portable endoscopy (YunSendo) for upper gastrointestinal endoscopy examination in Ba-Ma mini-pigs.Methods:A total of 10 Guangxi Ba-Ma mini-pigs were used in the experiment, and mucosal injury models were established in advance by biopsy forceps in esophagus, stomach, and duodenum. Each experimental animal underwent medical disposable portable endoscopy and Olympus endoscopy (GIF-Q260J) performed by two endoscopists separately. The time when the endoscope reached the duodenum, the number of detected mucosal injuries and endoscopic pictures of different parts with standard image acquisition were recorded. Endoscopic operational performance and endoscopic image quality were evaluated. Different endoscopists recorded experimental results with blind method. The procedures of the two endoscopic examinations were performed by coin-tossing method. The paired t test was used for statistical analysis. Results:There were no statistically significant differences in the insertion time and total operation time between medical disposable portable endoscopy and Olympus endoscopy ( (171.00±9.96) s vs. (164.00±17.84) s, (285.00±33.94) s vs. (273.40±23.46) s; t=1.289 and 1.281, P=0.230 and 0.232). There were no statistically significant differences in the percentage of time of clear visual field during endoscopy insertion and total operation between medical disposable portable endoscopy and Olympus endoscopy ((91.83±1.85)% vs. (91.52±1.51)%, (93.07±3.10)% vs. (92.06±2.57)%; t=0.401 and 0.689, P=0.698 and 0.508). Moreover, there were no statistically significant differences in the score of comprehensive operation performance, score of clear image number, score of image color recognition, score of image illumination, comprehensive score of image quality and number of detected mucosal injuries ((9.66±0.30) points vs. (9.86±0.15) points, (39.50±0.71) points vs. (39.30±1.06) points, (39.70±0.48) points vs. (39.40±0.70) points, (39.40±0.70) points vs. (39.50±0.71) points, (9.88±0.09) points vs. (9.85±0.20) points, 9.80±0.42 vs. 9.90±0.32; t=2.176, 1.000, 1.152, 0.317, 0.629 and 0.557, all P>0.05). There were no adverse events after operation in medical disposable portable endoscopy group and Olympus endoscopy group. Conclusions:The medical disposable portable endoscopy is safe and feasible for endoscopy examination in live animal models. Different parts of upper gastrointestinal tract and mucosal lesions can be clearly detected. The operational performance and the image quality are excellent, which is similar to Olympus endoscopy (GIF-Q260J).