OBJECTIVE To standardize the main processes and related technical links of the clinical comprehensive evaluation of drugs， and provide guidance and reference for improving the quality of comprehensive evaluation evidence and its transformation and application value. METHODS The construction of Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs was based on the standard guideline formulation method of the World Health Organization （WHO）， strictly followed the latest definition of guidelines by the Institute of Medicine of the National Academy of Sciences of the United States， and conformed to the six major areas of the Guideline Research and Evaluation Tool Ⅱ. Delphi method was adopted to construct the research questions； research evidence was established by applying the research methods of evidence-based medicine. The evidence quality classification system of the Chinese Evidence-Based Medicine Center was adopted for evidence classification and evaluation. The recommendation strength was determined by the recommendation strength classification standard formulated by the Oxford University Evidence-Based Medicine Center， and the recommendation opinions were formed through the expert consensus method. RESULTS & CONCLUSIONS The Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs covers 4 major categories of research questions， including topic selection， evaluation implementation， evidence evaluation， and application and transformation of results. The formulation of this guideline has standardized the technical links of the entire process of clinical comprehensive evaluation of drugs， which can effectively guide the high-quality and high-efficient development of this work， enhance the standardized output and transformation application value of evaluation evidence， and provide high-quality evidence support for the scientific decision-making of health and the rationalization of clinical medication.

Currently,human health due to corona virus disease 2019(COVID-19)pandemic has been seriously threatened.The coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein plays a crucial role in virus transmission and several S-based therapeutic approaches have been approved for the treatment of COVID-19.However,the efficacy is compromised by the SARS-CoV-2 evolvement and mutation.Here we report the SARS-CoV-2 S protein receptor-binding domain(RBD)inhibitor licorice-saponin A3(A3)could widely inhibit RBD of SARS-CoV-2 variants,including Beta,Delta,and Omicron BA.1,XBB and BQ1.1.Furthermore,A3 could potently inhibit SARS-CoV-2 Omicron virus in Vero E6 cells,with EC50 of 1.016 pM.The mechanism was related to binding with Y453 of RBD deter-mined by hydrogen-deuterium exchange mass spectrometry(HDX-MS)analysis combined with quan-tum mechanics/molecular mechanics(QM/MM)simulations.Interestingly,phosphoproteomics analysis and multi fluorescent immunohistochemistry(mIHC)respectively indicated that A3 also inhibits host inflammation by directly modulating the JNK and p38 mitogen-activated protein kinase(MAPK)path-ways and rebalancing the corresponding immune dysregulation.This work supports A3 as a promising broad-spectrum small molecule drug candidate for COVID-19.

It is an urgent demand worldwide to control the coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The 3-chymotrypsin-like protease (3CL^pro) and papain-like protease (PL^pro) are key targets to discover SARS-CoV-2 inhibitors. After screening 12 Chinese herbal medicines and 125 compounds from licorice, we found that a popular natural product schaftoside inhibited 3CL^pro and PL^pro with IC₅₀ values of 1.73 ± 0.22 and 3.91 ± 0.19 μmol/L, respectively, and inhibited SARS-CoV-2 virus in Vero E6 cells with EC₅₀ of 11.83 ± 3.23 μmol/L. Hydrogen-deuterium exchange mass spectrometry analysis, quantum mechanics/molecular mechanics calculations, together with site-directed mutagenesis indicated the antiviral activities of schaftoside were related with non-covalent interactions with H41, G143 and R188 of 3CL^pro, and K157, E167 and A246 of PL^pro. Moreover, proteomics analysis and cytokine assay revealed that schaftoside also regulated immune response and inflammation of the host cells. The anti-inflammatory activities of schaftoside were confirmed on lipopolysaccharide-induced acute lung injury mice. Schaftoside showed good safety and pharmacokinetic property, and could be a promising drug candidate for the prevention and treatment of COVID-19.

Consecutively hospitalized patients with confirmed coronavirus disease 2019 (COVID-19) in Wuhan, China were retrospectively enrolled from January 2020 to March 2020 to investigate the association between the use of renin-angiotensin system inhibitor (RAS-I) and the outcome of this disease. Associations between the use of RAS-I (angiotensin-converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB)), ACEI, and ARB and in-hospital mortality were analyzed using multivariate Cox proportional hazards regression models in overall and subgroup of hypertension status. A total of 2771 patients with COVID-19 were included, with moderate and severe cases accounting for 45.0% and 36.5%, respectively. A total of 195 (7.0%) patients died. RAS-I (hazard ratio (HR)= 0.499, 95% confidence interval (CI) 0.325-0.767) and ARB (HR = 0.410, 95% CI 0.240-0.700) use was associated with a reduced risk of all-cause mortality among patients with COVID-19. For patients with hypertension, RAS-I and ARB applications were also associated with a reduced risk of mortality with HR of 0.352 (95% CI 0.162-0.764) and 0.279 (95% CI 0.115-0.677), respectively. RAS-I exhibited protective effects on the survival outcome of COVID-19. ARB use was associated with a reduced risk of all-cause mortality among patients with COVID-19.
Angiotensin Receptor Antagonists/therapeutic use* ; Angiotensin-Converting Enzyme Inhibitors/therapeutic use* ; COVID-19 ; Humans ; Hypertension/drug therapy* ; Renin-Angiotensin System ; Retrospective Studies

Objective:To investigate the current status of care activities and supportive behaviors for family caregivers of patients with diabetic kidney disease and to analyze the influencing factors.Methods:A total of 280 family caregivers for diabetic kidney disease were chosen by convenience sampling in a Nephrology Ward of a ClassⅢ Grade A general hospital and a Diabetic Nephropathy Department of a diabetes specialist hospital in Tianjin from February to October 2019. The general information questionnaire, Barthel Index, Diabetes Caregiver Activity and Support Scale, Social Support Rating Scale, General Self-Efficacy Scale, Zarit Caregiver Burden Interview Scale were applied to conduct an investigation, and to explore the factors affecting the caring activities and supportive behaviors of family caregivers of patients with diabetic kidney disease.Results:The total score of caring activities and supporting behaviors of 280 family caregivers of patients with diabetic kidney disease was (61.23±8.07) . Multiple stepwise linear regression analysis showed that patients' self-care ability, family caregivers' self-efficacy, social support, caring burden, gender, education level were the factors influencing the caring activities and supportive behaviors of family caregivers (adjusted R2=0.558, F=59.769, P<0.001) . Conclusions:Family caregivers need to improve their caring activities and supportive behaviors further, and clinical staff should pay more attention to patients with following features: poor self-care abilities, low self-efficacy, low levels of social support, heavy caring burdens, female, and low levels of education, as well as to provide health education and training from the perspective of family caregivers to assist them in improving their caring activities and supportive behaviors.

Lycopene, as a high value-added terpene compound, has been widely concerned by researchers at home and abroad. Firstly, the ability of lycopene synthesis of Saccharomyces cerevisiae model strains S288c and YPH499 was analyzed and compared. The results showed that YPH499 was more suitable for lycopene synthesis as yeast chassis. Subsequently, the effects of constitutive promoters GPDpr, TEF1pr and inducible promoters GAL1pr, GAL10pr on Lycopene synthesis were compared. The results showed that when GPDpr and TEF1pr were used as promoters of crtE, crtB and crtI in lycopene synthesis pathway, the production of lycopene was 15.31 mg/L after 60 h fermentation in shaking flask. When GAL1pr and GAL10pr were used as promoters, the production was 123.89 mg/L, which was 8.09 times higher. In addition, the methylvaleric acid (MVA) pathway was further modified to overexpress the key enzyme gene of N-terminal truncation, tHMG1 (3-hydroxy-3-methylglutaryl coenzyme A reductase). The lycopene production was 265.68 mg/L, and the yield per cell was 72.79 mg/g. The Saccharomyces cerevisiae strain designed and constructed in this study can express lycopene in high yield per cell, thus could be used in the industrial production of lycopene after further construction and optimization.
Biosynthetic Pathways ; genetics ; Fermentation ; Industrial Microbiology ; Lycopene ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Species Specificity

Objective:To investigate the protective effect of ulinastatin combined with adipose-derived stem cells (ADSCs) transplantation on renal tissue in rats with endotoxic shock.Methods:20 Sprague Dawley rats were randomly selected as normal group from 108 Sprague Dawley rats. The remaining 88 rats were treated with 2 ml lipopolysaccharide (10 mg/kg) via tail intravenous injection to establish endotoxic shock model. The established 80 model rats were randomly divided into model group, ADSCs group, ulinastatin group and combination group (ulinastatin combined with ADSCs). All the rats were treated once a day for 3 days. Three days after transplantation, the renal tissues of each group were stained with hematoxylin-eosin staining to observe the pathological changes. The distribution of CM-Dil labeled ADSCs in the kidney of rats was observed by fluorescence microscope. The levels of nitric oxide synthase (NOS), nitric oxide (NO), creatinine and urea nitrogen in rat serum were measured. The reverse transcription PCR and Western Blot were used to detect the level of Bax and Caspase-3 in rat kidney tissue. Three days after transplantation, inflammatory cell infiltration and necrosis were occasionally seen in model group. Compared with the model group, the kidney damage in the ADSCs group and the ulinastatin group was significantly reduced, and kidney damage in the combined group was the least.Results:CM-Dil-labeled positive cells were found by microscope in the ADSCs group and the combined group, while CM-Dil-labeled ADSCs were not found in the kidney tissues of the normal group, model group and ulinastatin group. Compared with the normal group, the levels of NOS, NO, creatinine and urea nitrogen in the model group were significantly increased (all P<0.05). Compared with the model group, the levels of NOS, NO, creatinine and urea nitrogen in the ADSCs group, the ulinastatin group and the combination group were significantly increased (all P<0.05), in which the combined group has a further reduction in these related protein levels than the ADSCs group and the Ulinastatin group (all P<0.05). The mRNA and protein expressions of Bax and Caspase-3 in the kidney tissues of the model group were significantly higher than those in the normal group (all P<0.05). The Bax and Caspase levels in the kidney tissues of the ADSCs group, Ulinastatin group and combination group -3 mRNA and protein expression were significantly lower than those of the model group (all P<0.05), in which the combined group has a further reduction in these related protein levels than the ADSCs group and the Ulinastatin group (all P<0.05). Conclusions:Ulinastatin combined with ADSCs transplantation has a protective effect on kidney damage caused by endotoxin shock, which may be related to alleviating renal cell injury.

Objective: To explore the value of chest CT features and clinical indexes in the differential diagnosis between suspected COVID-19 with two or more negative nucleic acid tests and confirmed COVID-19. Methods: The clinical data and chest CT images of 105 cases withconfirmedCOVID-19 (55 males and 50 females, aged from 2 month to 88 years) and 97 cases with suspected COVID-19(59 males and 38 females, aged from 1 month to 93 years) were analyzed retrospectively in Shiyan Taihe Hospital from January 21 to February 10, 2020. χ²test and two independent sample t test were used to analyze the clinical data and CT signs of the two cases, with P<0.05 for the difference statistically significant. Results: Compared with the suspected patients, the average age of diagnosis of covid-19 was higher (t = 2.460, P = 0.01). The main pathological changes were pure ground glass (68 cases) and mixed ground glass density (53 cases) (χ² = 50.016, P< 0.01). Interstitial thickening (83 cases) (χ² = 55.395, P< 0.01), vascular thickening (73 cases) (χ² = 57.527, P< 0.01), air bronchoscopic sign or bronchiectasis Zhang (67 cases) (χ² = 17.899, P< 0.01), cord focus (54 cases) (χ² = 5.500, P = 0.02), easily distributed under the pleura and the long axis of the lesion was parallel to the pleura (89 cases) (χ² = 23.597, P< 0.01), most of them had no pleural effusion (χ² = 7.017, P< 0.01); both lesions were mainly distributed in patches (89 cases were confirmed, 87 suspected) (χ² = 19.573, P< 0.01). In addition, the lesions of patients with confirmed covid-19 showed progress in short term (72 / 87, 82.76%), and those with suspected covid-19 showed remission in short term (63 / 89, 70.78%). The difference was statistically significant (χ² = 51.114, P< 0.01). There was no significant difference in gender and distribution of pulmonary lobes (χ² = 1.462, P= 0.23; χ² = 7.381, P= 0.19). The number of white blood cells (χ² = 17.891, P< 0.01) and the percentage of lymphocytes (χ² = 11.151,P< 0.01) of covid-19 were mostly normal or decreased, creatine kinase (χ² = 9.589, P< 0.01) were mostly normal, and erythrocyte sedimentation rate was mostly normal or increased (χ² = 4.240, P= 0.04). Conclusions The imaging features and biochemical indexes of diagnosed COVID-19 are different from those of suspected COVID-19. The comparative analysis of imaging features, clinical indexes and reexamination are helpful for the differential diagnosis of COVID-19 and suspected COVID-19.

Objective To investigate the effects of tea-polyphenols on diabetic nephropathy (DN) mice by regulating nuclear factor E2-related factor 2/antioxidant response element (Nrf-2/ARE) signaling pathway. Methods A total of ten male 9-week-old normal (db/m) mice were randomly and equally divided into blank control group and tea-polyphenol control group, and ten male 9-week-old homologous type 2 diabetes (db/db) mice were randomly divided into model group and tea polyphenol treatment group. The animals in the tea-polyphenol control group and the treatment group were given 50 mg/(kg·d) tea-polyphenols by oral gavage, and the animals in the blank control group and model group were given same volume of double distilled water. The administration was once a day for 8 weeks. The blood glucose and 24-hour urine protein quantization (24 h-UP) were measured and recorded at 0, 4, and 8 weeks. After 8 weeks of the treatment, the mice were sacrificed. The intraocular blood stasis samples were collected for renal function indicators (serum creatinine and urea nitrogen), and kidney tissue samples were also collected for the tests of superoxide dismutase (SOD), reactive oxygen species, and malondialdehyde. Periodic acid Schiff reaction (PAS) staining was used to observe glomerular injury and scored. Western blot was used to detect the expression of Nrf-2 and hemeoxygenase-1 (HO-1) protein. Results Compared with the blank control group, the blood glucose and 24 h-UP of the mice in the model group and the tea-polyphenol treatment group increased after 4 and 8 weeks of the treatment (all P<0.01). Compared with the blank control group, after 8 weeks of the treatment, the serum creatinine and blood urea nitrogen of the model group and the tea-polyphenol treatment group increased (all P<0.01), the content of SOD in the renal tissue decreased (all P<0.01), the content of active oxygen and malondialdehyde, the relative expression of Nrf-2 and HO-1 protein increased (all P<0.01), and the glomerular injury aggravated (all P<0.01). However, there were no significant differences in all the indexes between the tea-polyphenol control group and the blank control group (all P>0.05). Conclusions Renal tissue of DN mice will undergo significant oxidative stress injury. Tea-polyphenols may reduce the oxidative stress injury in DN mice by regulating the Nrf-2/ARE signaling pathway, and play a protective role in the kidney.

Animals ; Cells, Cultured ; Gastrointestinal Tract ; metabolism ; Neuroglia ; metabolism ; Neurons ; metabolism ; Peptide Fragments ; metabolism ; Peptides ; metabolism ; Proteins ; metabolism ; Rats