Abstract] Objective To investigate the in vivo therapeutic effects of an extract of herb medi-cines, YiGanQingDuKeLi, in combination with adefovir dipivoxil (ADV) on the rebound of duck hepatitis B virus ( DHBV) multiplication after withdrawal of ADV treatment.Methods Peking ducks were infected with DHBV positive serum samples for 7 days and then screened by SYBR Green real-time PCR.The ducks positive for DHBV were randomly divided into five groups including the model control group, the ADV treat-ment group, the herb treatment group, the high-dose combination therapy group and the low-dose combina-tion therapy group.The ducks in the ADV treatment and the herb treatment groups were respectively treated with distilled water and YiGanQingDuKeLi (1.2 g/ml) for 14 days after the treatment of ADV (0.25 mg/ml) for 21 days.The ducks in the high-dose group were treated with YiGanQingDuKeLi (1.2 g/ml) for 14 days after the combined treatment with high-dose YiGanQingDuKeLi (1.2 g/ml) and ADV (0.25 mg/ml) for 21 days.The ducks in the low-dose group were treated with YiGanQingDuKeLi (0.6 g/ml) for 14 days after the combined treatment with YiGanQingDuKeLi (0.6 g/ml) and ADV (0.125 mg/ml) for 21 days.Blood samples were collected from each duck via leg vein after 0, 7, 14 and 21 days of drug adminis-tration and after 7 and 14 days of drug withdrawal.The levels of DHBV-DNA, alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) in blood serum samples were detected.Results Compared with the model group, the levels of DHBV-DNA, ALT and AST in ducks from the herb treatment group and combined treatment groups were decreased before the discontinuation of ADV treatment ( P<0.05 or P<0.01).Moreover, the titers of DHBV-DNA in ducks treated with high doses of drugs were much lower than those from ADV treatment group.The levels of DHBV-DNA, ALT and AST in ducks treated with herb medi-cine and high doses of drugs remained at relatively low levels after the cessation of ADV treatment, but re-bounded significantly in ducks with ADV treatment.The levels of DHBV-DNA and ALT rebounded slightly in ducks treated with low doses of drugs as compared with those of ADV treatment group ( P<0.01 or P<0.05).Conclusion The treatment of YiGanQingDuKeLi in combination with ADV could inhibit not only the in vivo replication of DHBV, but also the rebound of DHBV multiplication after ADV withdrawal.

Objective To investigate the correlation between ACE gene polymorphisms and stroke of Han nationality people in Fangshan district of Beijing. Methods The Insertion/Deletion(ID) polymorphisms of ACE gene were detected in 63 patients with cerebral hemorrhage,and 713 patients with cerebral infarction and 235 health control by polymerase chain reaction(PCR). We observed the frequencies of genotype of deletion homozygote(DD),insertion homozygote(II) and insertion/deletion heterozygote (ID) and the alleles of D and I. Also we analyzed the association among I/D polymorphisms of ACE gene with serum glucose(GLU),triglyceride(TG),cholesterol(TC) levels. Results There was no significant difference in the frequencies of both genotypes of DD,ID,II and alleles of D and I in three groups. The serum GLU levels in patients carrying ID,II genotype were higher than those in healthy control(P

Objective The aims of the study were to investigate the effects of human islet amyloid polypeptide (hIAPP) on autophagy in INS-1 cells and its underlying mechanism,and to explore the role of autophagy in hIAPP-induced cytotoxicity and oxidative stress.Methods INS-1 cells were treated with hIAPP (10 μmol/L) for 24 h in the presence or absence of N-acetyl-L-cysteine (NAC),compound C,5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) and 3-methyladenine (3-MA),respectively.Transmission electron microscopy was used to observe the number of autophagosome in cells.Cell viability was determined by methyl thiazolyl tetrazolium (MTT) test.2',7'-dichlorofluorescin diacetate (DCFH-DA) assay was used to measure the relative levels of reactive oxygen species (ROS).Western blot was used to detect expression of adenosine monophosphate-activated protein kinase (AMPK) and autophagic markers p62 and microtubule associated protein 1 light chain3 (LC3).Results Treatment of INS-1 cells with hIAPP resulted in a significant increase in the number of autophagosomes and the expression of LC3-Ⅱ/LC3-Ⅰ (both P＜0.05).Meanwhile,treatment of INS-1 cells with hIAPP enhanced the level of ROS to 1.76 times of control cells (P＜0.01).Co-treatment with NAC,an antioxidant,inhibited hIAPP-induced ROS generation,and the expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK in the INS-1 cells (all P＜0.05).Pretreatment of INS-1 cells with AMPK inhibitor compound C suppressed hIAPP and AICAR,an activator of AMPK,induced expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK (all P＜0.05).Autophagic inhibitor 3-MA and compound C aggravated the hIAPP-induced cell death and ROS generation in INS-1 cells (All P＜0.05).The cytotoxic effects of hIAPP were significantly attenuated by co-treatment with AICAR (P＜0.05).Conclusion Autophagy may act as an adaptive mechanism to alleviate hIAPP-induced oxidative damage and toxicity in INS-1 cells.

Objective The DNA methylation microhaplotype(DMH)refers to the combination of multiple methylation sites within a very short range,and these haplotypes show wide diversity.We carried out screening and validation of age-related DMHs in mouse blood.Methods We initially constructed a theoretical dataset of DMHs based on the mouse reference genome.We then screened age-related DMHs by Spearman's rank correlation analysis,using high-throughput sequencing information for DNA methylation in mouse blood from a network database.Finally,cross-validation was performed using a validation dataset.Results A total of 6787 142 DMH sites were identified within 50 bp in the mouse genome,including 98.64%of single-digit CpG sites.A total of 5835 age-associated DMHs were screened in 58 mouse blood samples(|rho|＞0.5,P＜0.01),accounting for 0.086%of DMHs.Finally,we validated the top 100 age-associated DMHs with high correlation in 95 independent samples,Resultsing in 44 loci.Conclusions The age-associated DMHs screened in this study may be useful in future studies of apparent age prediction using mouse blood and in aging studies.

Objective To explore the effect of low-level laser therapy (LLLT)applied to the quadriceps muscle on the recovery of exhausting-cycling-exercise-induced fatigue.Methods According to a randomised,double-blind and crossover design,16 healthy male students were randomly assigned to an LLLT-1,LLLT-3,LLLT-5 and a placebo group,and received LLLT for 300 s at the dosage of 0.06 J· cm-2,0.18 J·cm-2,0.3 J·cm-2 and 0 to the bilateral rectus femoris after the exhausting-cycling-exercise-induced fatigue.The blood lactate(BL),heart rate(HR),rated perceived exertion(RPE)and visual analogue scale(VAS)were assessed before the exercise,immediately after exercise,10 and 20 min after exercise,as well as immediately after the first Wingate(WG)test,5 and 30 min after the WG test.Meanwhile,the second WG test was performed 40 min after the first WG test.Results The average HR value of LLLT-1 group was significantly lower than the placebo group at 10 min after exercise(P＜ 0.05)and immediately after the WG test(P＜0.01),while that of LLLT-3 and LLLT-5 groups was significantly lower than the placebo group immediately and 5 min after the WG test(P＜0.01).Compared to the placebo group,the average BL of LLLT-1,LLLT-3 and LLLT-5 groups was significantly lower 10 min after exercise(P＜0.05 for all)and that of LLLT-5 group was also significantly lower 30 min after the first WG test(P＜0.05).However,the average blood glucose of LLLT-5 group was significantly higher than the placebo group right after the first WG test(P＜0.05).Moreover,significant increase was observed in the mean(P=0.002)and peak power(P=0.006)at the second WG test and the mean(P=0.048) power at the first WG test of LLLT-3 group,compared to the placebo group.Conclusion LLLT applied to quadriceps muscles after exhausting exercise may enhance recovery,and LLLT at the dose of 0.18 J·cm-2 is of the best effect.

Objective: To explore an effective long non-coding RNA (lncRNA) signature in predicting the prognosis of hepatocellular carcinoma through the analysis on RNA sequencing data of hepatocellular carcinoma patients and peritumoral tissues in the Cancer Genome Atlas (TCGA) database. Methods: The clinical characteristics and RNA sequencing data of 377 hepatocellular carcinoma patients were obtained from TCGA database by the end of February 2018. Then, differentially expressed lncRNAs between 50 pairs of tumor and peritumoral tissues were explored using student’s t-test. Next, a lncRNA signature was established through LASSO Cox regression analysis. All the patients were divided into four groups (<P₂₅, P₂₅-, P₅₀-, ≥P₇₅) based on the cut-off quartiles signature. Finally, compared with the control group (<P₂₅), the hazard ratios (HRs) of three groups (P₂₅-, P₅₀-, ≥P₇₅) were calculated by using Cox regression. The survival outcomes of patients in the four groups were compared to evaluate the capacity of the lncRNA signature model. Results: A total of 951 differentially expressed lncRNAs were identified between tumor and peritumoral tissues. A three-lncRNA signature, including LNCSRLR, MKLN1-AS and ZFPM2-AS1, was established to predict the prognosis of hepatocellular carcinoma patients. The outcome suggested that the death risk of the ≥P₇₅ group was 1.57 times larger than that of the <P₂₅ group (95%CI: 1.06-2.31, P<0.05). Conclusion The three-lncRNA signature, which established by LNCSRLR, MKLN1-AS and ZFPM2-AS1, was significantly associated with the prognosis of hepatocellular carcinoma patients based on TCGA database data.

Objective:To explore the efficacy and safety of sivelestat, a neutrophil elastase (NE) inhibitor, in the treatment of acute lung injury (ALI) in the intensive care unit (ICU).Methods:A retrospective analysis was performed on 171 patients with ALI in the ICU of the First Affiliated Hospital of Zhengzhou University from June 2020 to June 2021, including 77 patients in the sivelestat group and 94 patients in the conventional treatment group. Acute physiology and chronic health evaluation (APACHE) Ⅱ score, Murray lung injury score, oxygenation index (PaO 2/FiO 2 ratio), inflammatory cytokines (IL-6, IL-10, TNF-α), ventilator-free days (VFD), the length of ICU stay, and the 28-day mortality were collected to assess the efficacy of sivelestat. At the same time, adverse reactions and laboratory test results within 30 days after the use of sivelestat were recorded to assess the safety. Results:Compared with conventional treatment, oxygenation index, Murray lung injury scores, IL-6, IL-10, and TNF-α were significantly improved after 7 days of sivelestat treatment. Compared with the conventional treatment group, the VFD was significantly longer ( P = 0.0119) and the length of ICU stay was significantly shorter ( P = 0.0269) in the sivelestat group. The mortality was 14.29% in the sivelestat group and 22.34% in the conventional treatment group and, with no statistically significant. In the meantime, sivelestat did not increase adverse reactions within 30 days after treatment. Conclusions:Sivelestat treatment is safe and more effective than conventional treatment for ALI patients in the ICU.

Objective:To investigate the effect and mechanism of isorhynchophylline (IRN) on angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy.Methods:H9c2 cells were co-cultured with Ang Ⅱ and different concentrations of IRN (0, 5, 10, 25, 50 μmol/L). The cell surface area and mRNA levels of cardiac hypertrophy markers atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) were detected to elucidate the effect of IRN on myocardial hypertrophy and the most effective concentration. H9c2 cells were co-cultured with Ang Ⅱ and IRN (25 μmol/L) at different times (0, 6, 12, 24 h) to elucidate the most effective time of inhibition. The phosphorylation levels of the signaling pathway were detected, and the effects of IRN and Akt inhibitor MK2206 on the phosphorylation levels of the signaling pathway were further explored to elucidate the underlying mechanisms.Results:Compared with the control group, the surface area of H9c2 cells, and the mRNA expression of myocardial hypertrophy markers ANP, BNP and β-MHC were significantly increased (all P<0.05). Pretreated with different concentrations of IRN (5, 10, 25, 50 μmol/L) could inhibit the increase in cell surface area induced by AngⅡ (all P<0.05), especially at the concentration of 25 μmol/ L ( P<0.01). IRN could time-dependently inhibit AngⅡ-induced activation of ANP, BNP, β-MHC mRNA (all P<0.05). AngⅡ caused increased phosphorylation levels of Akt, GSK3β, mTOR and FOXO3a. IRN could block AngⅡ-induced phosphorylation of the Akt signaling pathway. Conclusion:IRN attenuates AngⅡ-induced cardiomyocyte hypertrophy by inhibiting the Akt signaling pathway.

Objective:To investigate the clinicopathological characteristics of the T cell lymphomas with CD20 expression, and to better understand this rare entity.Methods:Two-hundred cases of T-cell lymphoma diagnosed in the Department of Pathology of the Affiliated Hospital of Qingdao University from November 2016 to February 2020 were examined, and 5 cases of CD20-positive T-cell lymphomas were identified and included. Combined with clinical data and review of the literature, the clinicopathological characteristics of the disease were analyzed.Results:The five patients were all male, and had an average age of 56 years (range, 47 to 64 years). There were 2 cases of monomorphic epitheliotropic intestinal T-cell lymphoma, 2 cases of mycosis fungoides (1 case was plaque stage and the other was tumor stage) and 1 case of indolent T-cell lymphoproliferative disorder of the gastrointestinal tract. Immunohistochemistry showed that all 5 cases expressed multiple T cell markers (CD3/CD4/CD5/CD7/CD8) and only one of B cell markers (CD20). Three of the 5 cases were negative for CD20 at the first diagnosis, while CD20 was diffusely positive on the second biopsy from the recurrence or progression of the disease, without expression of CD79a or PAX5. Epstein-Barr encoding region (EBER) in situ hybridization was negative in all 5 cases. T-cell receptor gene analysis showed monoclonal rearrangement of β or/and δ chains;Ig rearrangements were all polyclonal. None of the five patients were treated with rituximab, and 4 patients survived with disease and 1 patient survived without disease at the end of follow-up. Among them, the patient with mycosis fungoides at the cancerous stage has progressed rapidly and had poor quality of life.Conclusions:CD20-positive T-cell lymphoma is extremely rare. Its prognosis is closely related to the type of T-cell lymphoma, clinical stage and initial therapeutic effect. However, the expression of CD20 indicates the recurrence or progression of the disease, and the prognosis is relatively poor. When CD3 expression is absent in T-cell lymphoma, it is easy to be misdiagnosed as B-cell lymphoma. The combination of multiple immunohistochemical antibodies and molecular detection can improve the accuracy of diagnosis.