Active Ingredients of Reduning Injection Maintain High Potency against SARS-CoV-2 Variants / 中国结合医学杂志

OBJECTIVE@#To investigate the anti-coronavirus potential and the corresponding mechanisms of the two ingredients of Reduning Injection: quercetin and luteolin.@*METHODS@#A pseudovirus system was designed to test the efficacy of quercetin and luteolin to inhibit SARS-CoV-2 infection and the corresponding cellular toxicity. Luteolin was tested for its activities against the pseudoviruses of SARS-CoV-2 and its variants. Virtual screening was performed to predict the binding sites by Autodock Vina 1.1.230 and PyMol. To validate docking results, surface plasmon resonance (SPR) was used to measure the binding affinity of the compounds with various proteins of the coronaviruses. Quercetin and luteolin were further tested for their inhibitory effects on other coronaviruses by indirect immunofluorescence assay on rhabdomyosarcoma cells infected with HCoV-OC43.@*RESULTS@#The inhibition of SARS-CoV-2 pseudovirus by luteolin and quercetin were strongly dose-dependent, with concentration for 50% of maximal effect (EC50) of 8.817 and 52.98 µmol/L, respectively. Their cytotoxicity to BHK21-hACE2 were 177.6 and 405.1 µmol/L, respectively. In addition, luetolin significantly blocked the entry of 4 pseudoviruses of SARS-CoV-2 variants, with EC50 lower than 7 µmol/L. Virtual screening and SPR confirmed that luteolin binds to the S-proteins and quercetin binds to the active center of the 3CLpro, PLpro, and helicase proteins. Quercetin and luteolin showed over 99% inhibition against HCoV-OC43.@*CONCLUSIONS@#The mechanisms were revealed of quercetin and luteolin inhibiting the infection of SARS-CoV-2 and its variants. Reduning Injection is a promising drug for COVID-19.

Positive Association of Human SHC3 Gene with Schizophrenia in a Northeast Chinese Han Population

Objective@#Schizophrenia is one of the most devastating neuropsychiatric disorders. Genetic epidemiological studies have confirmed that schizophrenia is a genetic disease. Genes promoting neurodevelopment may be potential candidates for schizophrenia. As an adaptor linking a number of tyrosine kinase receptors in multiple intracellular signaling cascades, Src homology 2 domain containing transforming protein 3 (SHC3) is a member of the Shc-like adaptor protein family, and expressed predominantly in the mature neurons of the central nervous system (CNS). In the present study, we aimed to investigate the association of SHC3 and schizophrenia. @*Methods@#An independent case-control association study was performed in a sample including 710 schizophrenia patients and 1314 healthy controls from a Northeast Chinese Han population. @*Results@#The allelic and genotypic association analyses showed that four SNPs in SHC3 significantly associated with schizophrenia (rs2316280, rs4877041, rs944485 and rs7021743). The haplotype composing of these four SNPs also showed significantly individual and global association with schizophrenia. @*Conclusion@#Our present results suggest SHC3 as a susceptibility gene for schizophrenia.

In Silico Screening of Potential Spike Glycoprotein Inhibitors of SARS-CoV-2 with Drug Repurposing Strategy / 中国结合医学杂志

OBJECTIVE@#To select potential molecules that can target viral spike proteins, which may potentially interrupt the interaction between the human angiotension-converting enzyme 2 (ACE2) receptor and viral spike protein by virtual screening.@*METHODS@#The three-dimensional (3D)-coordinate file of the receptor-binding domain (RBD)-ACE2 complex for searching a suitable docking pocket was firstly downloaded and prepared. Secondly, approximately 15,000 molecular candidates were prepared, including US Food and Drug Administration (FDA)-approved drugs from DrugBank and natural compounds from Traditional Chinese Medicine Systems Pharmacology (TCMSP), for the docking process. Then, virtual screening was performed and the binding energy in Autodock Vina was calculated. Finally, the top 20 molecules with high binding energy and their Chinese medicine (CM) herb sources were listed in this paper.@*RESULTS@#It was found that digitoxin, a cardiac glycoside in DrugBank and bisindigotin in TCMSP had the highest docking scores. Interestingly, two of the CM herbs containing the natural compounds that had relatively high binding scores, Forsythiae fructus and Isatidis radix, are components of Lianhua Qingwen (), a CM formula reportedly exerting activity against severe acute respiratory syndrome (SARS)-Cov-2. Moreover, raltegravir, an HIV integrase inhibitor, was found to have a relatively high binding score.@*CONCLUSIONS@#A class of compounds, which are from FDA-approved drugs and CM natural compounds, that had high binding energy with RBD of the viral spike protein. Our work provides potential candidates for other researchers to identify inhibitors to prevent SARS-CoV-2 infection, and highlights the importance of CM and integrative application of CM and Western medicine on treating COVID-19.

In Silico Screening of Potential Spike Glycoprotein Inhibitors of SARS-CoV-2 with Drug Repurposing Strategy / 中国结合医学杂志

OBJECTIVE@#To select potential molecules that can target viral spike proteins, which may potentially interrupt the interaction between the human angiotension-converting enzyme 2 (ACE2) receptor and viral spike protein by virtual screening.@*METHODS@#The three-dimensional (3D)-coordinate file of the receptor-binding domain (RBD)-ACE2 complex for searching a suitable docking pocket was firstly downloaded and prepared. Secondly, approximately 15,000 molecular candidates were prepared, including US Food and Drug Administration (FDA)-approved drugs from DrugBank and natural compounds from Traditional Chinese Medicine Systems Pharmacology (TCMSP), for the docking process. Then, virtual screening was performed and the binding energy in Autodock Vina was calculated. Finally, the top 20 molecules with high binding energy and their Chinese medicine (CM) herb sources were listed in this paper.@*RESULTS@#It was found that digitoxin, a cardiac glycoside in DrugBank and bisindigotin in TCMSP had the highest docking scores. Interestingly, two of the CM herbs containing the natural compounds that had relatively high binding scores, Forsythiae fructus and Isatidis radix, are components of Lianhua Qingwen (), a CM formula reportedly exerting activity against severe acute respiratory syndrome (SARS)-Cov-2. Moreover, raltegravir, an HIV integrase inhibitor, was found to have a relatively high binding score.@*CONCLUSIONS@#A class of compounds, which are from FDA-approved drugs and CM natural compounds, that had high binding energy with RBD of the viral spike protein. Our work provides potential candidates for other researchers to identify inhibitors to prevent SARS-CoV-2 infection, and highlights the importance of CM and integrative application of CM and Western medicine on treating COVID-19.

Effect of Norvasc and Tanatril on circadian rhythm of blood pressure in hypertensive patients / 中国地方病学杂志

Objective To observe the circadian rhythm of blood pressure and investigate the impact of Norvasc and Tanatril administrated at different time points based on the theory of time therapeutics in hypertensive patients. Methods The 24-hour ambulatory blood pressure was monitored in 130 inpatients and outpatients with grade 3 or 2 hypertension from May 2008 to November 2009. A total of 34 dipper hypertensive patients were grouped into dipper blood pressure group and 5 mg/d of Norvasc and Tanatril were taken by them at 6:00 AM. A total of 96 nondipper hypertensive patients were further divided into three subgroups after adjustment for age and gender: 5 mg/d of Norvasc and Tanatril were taken by group I (n = 30) at 6:00 AM; 5 mg/d of Tanatril at 6:00 AM and 5 mg/d of Norvasc at 18:00 PM by group Ⅱ (n = 32); 5 mg/d of Norvasc at 6:00 AM and 5 mg/d of Tanatrilat 18:00 PM by group Ⅲ(n=34).The 24-hour ambulatory blood pressure monitoring was performed again after four weeks treatment and 24-hour mean systolic blood pressure(24 hSBP),24-hour mean diastolic blood pressure(24 hDBP),daytime and nighttime mean systolic blood pressure(dSBP,nSBP)and daytime and nighttime mean diastolic blood pressure(dDBP,nDBP),were analyzed.Results The 24 hSBP,24 hDBP and dSBP,and dDBP were reduced from(154.3±5.6),(95.4±3.1),(158.7±6.5),(99.6±3.7)mmHg to(137.2±3.9),(82.5±2.7),(139.7±3.8),(85.2±3.5)mmHg,respectively,in dipper blood pressure group(t=2.124,2.356,2.278,2.449,all P<0.05).The 24 hSBP and 24 hDBP of the three groups in nondipper blood pressure were decreased from(154.4±6.1),(156.7±6.7),(156.6±5.2),(95.8±2.8),(94.9±3.8),(95.7±3.2)mmHg to (139.6±4.1),(134.5±4.6),(133.4±3.5),(83.5±4.2),(80.8±5.6),(81.6±4.7)mmHg,respectively(t=2.038,2.040,2.135,2.142,2.213,2.216,all P<0.05).dSBP and dDBP were decreased from(158.6±3.50),(158.4±5.6),(159.5±4.),(98.4±3.7),(99.6±3.7),(83.9±5.2)mmHg to(138.9±5.4),(136.7±4.1),(137.4±6.4),(85.8±5.3),(83.6±5.1),(83.9±5.2)mmHg,respectively(t=2.021,2.252,2.261,2.217,2.167,2.076,all P<0.05).nSBP and nDBP were decreased from(146.7±6.9),(149.8±3.9),(150.2±4.1),(93.7±4.2),(95.7±4.3),(93.4±3.3)mmHg to(133.7±4.6),(129.8±5.7),(127.6±2.8),(87.8±2.9),(78.5±6.4),(77.8±4.8)mmHg,respectively(t=1.798,2.032,2.014,1.864,2.157,2.166,all P<0.05).There were significant differences in nSBP and nDBP among all groups after treatment(F=2.32,2.17,all P<0.05),and the effect of the drugs was better in groug Ⅱ and Ⅲ than in group Ⅰ(q=3.17,4.03,3.32,4.19,all P<0.05),but there were no significant differences between group Ⅱ and Ⅲ(P>0.05).Conclusions Blood pressure can be controlled effectively by taking two antihypertensive medictions in the morning in hypertensive dipper patient but the blood pressure of nondipper hypertensive patients were able to be controlled more efficiently by taking the two antihypertensive medictions separately in the morning and at the evening compared with that taking the two drugs together in the morning.

Research of repairing rabbit knee joint cartilage defect by compound material of fibrin glue and decalcified bone matrix (DBM) and chondrocytes / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the feasibility and effectivity of using compound material of fibrin glue and DBM as scaffolds for cartilage tissue engineering.</p><p><b>METHODS</b>Chondrocytes isolated from articular cartilage were seeded into prepared scaffolds, after incubation for 4 weeks in vitro. Chondrocytes and fibrin glue and DBM constructs were implanted in the joint cave of rabbit. The specimens were excised at the 4th, 8th, 12th week, examined grossly analyzed by haematoxylin cosine, toluidine blues staining and type II collagen immunohistochemistry reaction. Wakitani score was counted to evaluate the repairing effect.</p><p><b>RESULTS</b>Grossly analysis showed some ivory tissue filled the caves after 4 weeks and the caves were full filled with smooth surface after 12 weeks. The microscope showed a good deal of chondrocytes appeared after 8 weeks and more type II collagen than 4 weeks. Twelve weeks later, cartilage lacuna could be observed. The cells arrangement and the amount of type II collagen both showed the same as the natural one.</p><p><b>CONCLUSION</b>Complicated material of fibrin glue and DBM as scaffolds can be used as scaffolds for cartilage tissue engineering.</p>

Berbamine induces apoptosis in human hepatoma cell line SMMC7721 by loss in mitochondrial transmembrane potential and caspase activation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To investigate the effect of berbamine on human hepatoma cell line SMMC7721.</p><p><b>METHODS</b>The effects of 24 h and 48 h incubation with different concentrations (0 to approximately 64 microg/ml) of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI (propidium iodide) staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential (psi(m)); the expression of activated caspase3 and caspase9 was analyzed by Western-blot.</p><p><b>RESULTS</b>The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential (psi(m)) and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk.</p><p><b>CONCLUSION</b>Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.</p>

Laparoscopic distal pancreatectomy with preservation of the spleen / 中华外科杂志

<p><b>OBJECTIVE</b>To summarize the experience of laparoscopic distal pancreatectomy with preservation of the spleen.</p><p><b>METHODS</b>From Nov 2003 to Dec 2004, 2 patients with cystic lesions in the body and tail of the pancreas underwent laparoscopic distal pancreatectomy with preservation of the spleen.</p><p><b>RESULTS</b>Both of the operations were successful with the operative time of 220 min and 190 min respectively, and the blood loss were 450 ml and 350 ml. There was no postoperative complication and the pathological diagnosis was pancreatic serous cystadenoma. Both patients' symptom disappeared after operation without recurrence during the follow-up of 18 and 5 months.</p><p><b>CONCLUSION</b>Laparoscopic distal pancreatectomy with preservation of the spleen is safe and feasible for the management of benign tumor in the body and tail of pancreas with the advantages of reduced injury, earlier recovery and less complication.</p>