Establishment and evaluation of a TaqMan RT-qPCR method for tissue quantification of oncolytic virus M1 in vivo / 中国病理生理杂志

AIM:To establish a TaqMan RT-qPCR method for surveiling the spread of oncolytic virus M1 in tissue,helping control the dosage and assessing the safety of virus. METHODS:A TaqMan-based one-step RT-qPCR method for the detection and quantification of oncolytic virus M1 in the tissues was established. The virus load and distri-bution in the tissues of SD rats,cynomolgus monkeys and nude mice were also investigated. RESULTS:A pair of specific primers(Q3)and the standard viral RNA for SYBR Green RT-qPCR were screened and selected with the best specificity and amplification efficiency. By optimizing the experiment conditions,we found that the annealing temperature above 62℃reduced matrix effect but affected the amplification efficiency. So we established a one-step TaqMan RT-qPCR method and redesigned a pair of Q3 short primers(Q3S). Using the one-step TaqMan RT-qPCR and Q3S primer,the stan-dard RNA with low copy numbers was specifically detected under the background of mixed matrix RNA of SD rats or cyno-molgus monkeys. Furthermore,the method was verified to be suitable for detecting tissue distribution of M1 virus in the mice,SD rats and cynomolgus monkeys. CONCLUSION:The TaqMan-based one-step RT-qPCR constructed with Q3S primer can be used for M1 virus quantification in various tissue samples of different animals with better specificity and sen-sitivity,and may be further applied to the detection of clinical samples.

Safety of protease inhibitors and Arbidol for SARS-CoV-2 pneumonia in Zhejiang Province, China / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The aim of this study was to evaluate the safety of an antiviral regimen of protease inhibitors combined with Arbidol (umifenovir) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pneumonia patients. The genomic sequence of SARS-CoV-2 is highly homologous to that of SARS-CoV (Zhou et al., 2020). Previously published basic and clinical research on anti-SARS-CoV treatment found that lopinavir/ritonavir (LPV/r) could improve the prognosis of SARS patients (Chan et al., 2003; Chu et al., 2004). Darunavir (DRV) is another protease inhibitor that blocks the binding of SARS-CoV-2 to human angiotensin-converting enzyme 2 (Omotuyi et al., 2020). The broad-spectrum antiviral drug Arbidol (umifenovir) also shows in vitro anti-SARS-CoV activity (Khamitov et al., 2008).

Retrospective Analysis of the Impact of the Adjuvant Huoxuehuayu Drugs on Upper Gastrointestinal Haemorrhage in Patients with Acute Coronary Syndrome / 中国药学杂志

OBJECTIVE: To analyze the usage of the adjuvant huoxuehuayu drugs in patients with acute coronary syndrome(ACS), observe the impact of the drugs on upper gastrointestinal haemorrhage(UGH), and provide a reference for the clinical rational use of the adjuvant huoxuehuayu drugs. METHODS: The ACS patients were enrolled in our hospital during May to July 2016. And the patients were divided into four groups according to whether using the adjuvant huoxuehuayu drugs: not use, used one kind, used two kinds, used ≥three kinds of huoxuehuayu drugs. Then we collected medical history, therapeutic measures and other baseline data, gathered the data of the usage of the adjuvant huoxuehuayu drugs, evaluated CRUSADE bleeding risk and analyzed frequency of occurrence of UGH. RESULTS: Overall 503 ACS patients were enrolled, there was no significant differences among the four groups when the medical history, essential medicines and intervene frequency were compared respectively. On the other hand, the incidence rate of UGH increased significantly with CRUSADE bleeding risk rank increasing incrementally.For very low and low bleeding risk ACS patients, there were no significant differences in UGH incidence rate among the four groups; For moderate and high bleeding risk ACS patients, UGH incidence rate increased significantly in ACS patients using more than one kind of huoxuehuayu drugs compared with other two groups(P<0.05). For very high bleeding risk ACS patients, UGH incidence rate increased significantly in ACS patients using 2 and ≥3 kinds of huoxuehuayu drugs compared with patients using one kind drug, P=0.009, 0.025 respectively. CONCLUSION: For moderate and high bleeding risk ACS patients, UGH incidence rate increase significantly in ACS patients using ≥2 kinds of huoxuehuayu drugs. A significant increase in the incidence rate of UGH with CRUSADE bleeding risk rank increasing, the use of huoxuehuayu drugs shoud be controled strictly, especially for very high bleeding risk ACS patients.

Research advance on Brucella Ⅳ secretion system proteins and its immune response / 中国人兽共患病学报

Brucella is a facultative intracellular pathogen that causes infection in domestic animal and humans,mainly parasitizing in cells and macrophages of hosts.Brucella can lead to abortion and sterility in animals.Meanwhile,it also causes arthralgia,weakness,undulat fever,hepatosplenomegaly and other symptoms in humans.Brucella relies on immune escape mechanism in the confrontation with the host.It can help Brucella "camouflage" to evade the identification of the immune system of the hosts and replicate within cells to persist to establish a long-term infection in the host.As a result of the existence of this mechanism,the treatment of Brucella infection is quite difficult.Type Ⅳ secretion system (T4SS) is a key virulence factor and it is essential for Brucella to survive in host cells.The effector proteins secreted by T4SS can help regulate the immune response against Brucella.In this article,we reviewed the studies on related proteins of the type Ⅳ secretion system of Brucella and its immune response,especially the relationship between the secretions of effector proteins mediated by VirB operon and immunity of the host.

Preventive effect of senecio cannabifolius less.II on perfluoroisobutylene inhalation-induced acute lung injury in rats / 国际药学研究杂志

Objective To preliminarily investigate the effect and possible mechanisms of Senecio cannabifolius Less. II (FHC- n) on perfluoroisobutylene (PFIB) inhalation-induced acute lung injury. Methods Totally 156 rats were randomly assigned to three groups: the control group， the PFIB group and the FHC- H prevention group， with 32， 62 and 62 rats in each group respectively. The FHC- II prevention group were given FHC- II three times per day at the dosage of 340 mg/kg before PFIB exposure. 1 h after the last time of FHC- h administration，the FHC- i prevention group were exposured to gaseous PFIB (0.2 mg/L) for 10 minutes in a static whole-body exposure inhalation system. The survival rate of the rats were recorded at 1，2，4，8，16，24，48 and 72 h post PFIB exposure; the lung index and total protein content in bronchoalveolar lavage fluid (BALF) were measured at 1 h，2 h，4 h，8 h，16 h and 24 h; IL-1β and IL-8 in sera were assayed by enzyme-linked immunosorbent assay (ELISA) at 1，2，4，8 and 16 h post PFIB exposure and the histopathological examination of the lung tissue was performed at 8 h post PFIB exposure. Results FHC-II significantly reduced the content of the total protein in BALF，lung index and the levels of IL-1β and IL-8 in aera as well， and dramatically alleviated the histopathological changes in the lung tissue. Conclusion FHC- D demonstrates some preventive effect on PFIB inhalation-induced acute lung injury in rats.

Efficacy and safety of the treatment: combination of benazepril/lercanidipine vs. benazepril alone in patients with mild-to-moderate hypertension / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Combination therapy is an effective method to reduce the blood pressure (BP) for patients with hypertension. This study was performed to evaluate the efficacy and safety of benazepril/lercanidipine compared with benazepril alone in patients with mild-to-moderate hypertension.</p><p><b>METHODS</b>One hundred and eighty-one patients with mild-to-moderate primary hypertension were assigned in this randomized, single-blind, parallel-group study and were randomly divided into group A (benazepril 10 mg/lercanidipine 10 mg) and group B (benazepril 10 mg) for 8 weeks. At 4 weeks, the dosage of Benazepril was titrated up to 20 mg if the diastolic blood pressure (DBP) remained ≥ 90 mmHg. BP control and side effects were evaluated at the end of 1, 4 and 8 weeks.</p><p><b>RESULTS</b>The baseline characteristics of the two groups were similar. The BP in both groups decreased from the baseline (P < 0.05). At the end of 4 and 8 weeks, Benazepril/Lercanidipine produced greater BP reduction than Benazepril alone (P < 0.05). The comparison of the rate of BP control for the benazepril/lercanidipine and benazepril groups at the end of 1, 4, and 8 weeks were 41.2% vs. 37.6% (P > 0.05), 67.1% vs. 44.7% (P < 0.05), and 71.8% vs. 45.9% (P < 0.05). There was no significant difference of side effects between the two groups.</p><p><b>CONCLUSION</b>The benazepril/lercanidipine combination is more effective in reducing BP than benazepril alone, while it does not increase the incidence of side effects.</p>