Zhizichi Tang （栀子豉汤）， first recorded in Treatise on Febrile and Miscellaneous Diseases （《伤寒杂病论》） by ZHANG Zhongjing， a medical sage during the Han dynasty， is one of the classical prescriptions in traditional Chinese medicine （TCM）. It plays an important role in the clinical practice of TCM because of its dispersing and transparent characteristics. It is one of the representative parts of “dispersing fire stagnation” and is used mainly for the treatment of various symptoms caused by heat depression in the chest and diaphragm. Pharmacological research has found that it has multiple effects， such as sedative hypnosis and anti-depression， inhibiting oxidative stress and inflammatory responses， regulating the intestinal flora， improving insulin resistance and endocrine metabolism disorders， reducing liver toxicity， and protecting the nerve and heart. Clinical studies have confirmed that its treatment of anxiety， depression， insomnia， and other diseases has few side effects and high safety. Combined with the analysis of TCM syndrome and pharmacological effects， Zhizichi Tang also shows potential in treating other diseases such as heart， lung system， spleen and stomach， liver system， endocrine， and metabolic system diseases. Therefore， the authors， by searching Chinese and foreign literature， especially in recent five years， systematically reviewed and summarized the research progress on Zhizichi Tang in six aspects of TCM syndrome， dosage and administration， chemical composition， pharmacological effects， clinical application， and adverse reactions， aiming to provide a reference for further research and clinical application of Zhizichi Tang.

Objective To explore the clinical and imaging features of phosphaturic mesenchymal tumor(PMT).Methods The clinical presentations,laboratory examinations,and imaging manifestations of seven patients with PMT diagnosed by surgery and pathology were analyzed retrospectively.Results Among the 7 patients,four patients had clinical presentations of long-term fatigue and bone pain.All patients showed preoperative blood phosphorus reduction in varying degrees.X-ray examination showed systemic osteomalacia and osteoporosis,accompanied by multiple pathological fractures.On CT,the primary tumor appeared as a soft tissue density mass or a ground glass high-density nodule with irregular calcification and local bone destruction.MRI showed long T1,long T2 signal intensity,and irregular low signal foci were scattered in the T2WI fat-suppressed sequence.The enhanced scans showed moderate to significant inhomogeneous enhancement.One patient who underwent 18F-FDG PET/CT and two patients who underwent 18F-ALF-NOTA-Octreotide(18F-OC)PET/CT examinations showed varying degrees of radioactive concentration in the lesions.Conclusion The clinical presentations and laboratory examinations of patients with PMT have certain characteristics.Systemic osteomalacia with pseudofracture line,calcification matrix within the tumor,and significant inhomogeneous enhancement of the lesion are the key imaging features for diagnosing PMT.18F-OC PET/CT examination plays a crucial role in the systemic localization diagnosis of tumors.

Objective:A SARS-CoV-2 pseudovirus(PsV)system was established for neutralizing antibody evaluation and virus entry inhibitor screening.Methods:Lentiviral vector plasmids psPAX2,pCDH-Luc and SARS-CoV-2 Spike(S)protein expres-sion plasmids were co-transfected,and harvested pseudoviral supernatant was used to infect ACE2-293T cells.Protein content of p24 was determined to reflect titer of PsV,and expression of S protein in PsV was detected by Western blot.Neutralization capacity of an S protein monoclonal antibody was evaluated using original strain,D614G,Gamma,Delta,Omicron PsV.Two reported virus entry inhibitors,chloroquine and carrageenin,were used to detect effect on entry of Omicron PsV.Results:Lentiviral vector successfully incorporated S protein.Western blot results showed that S protein mutated at 665Y showed a different cleavage form(90 kD)than wild-type full-length S protein(180 kD).Titer of PsV packaged by three plasmids system was higher.Ratio of S protein expression plasmid,transfer plasmid and packaging plasmid at 1∶3∶3 was optimum condition for viral packaging.Titer of PsV packaged under this condi-tion was over 20 ng/ml.PsV could effectively infect ACE2-293T cells,and double reporter gene GFP and firefly luciferase were expressed obviously,whose chemiluminescence values reached 106.Monoclonal antibodies of S protein effectively neutralized four types of PsVs,but neutralization of original strain was 10～30 times greater than that of variant PsV.Virus entry inhibitors,chloroquine and ι-carrageenan significantly inhibited entry of Omicron PsV.Conclusion:SARS-CoV-2 PsV infection system we conducted can simu-late entry of SARS-CoV-2 successfully.Effective pharmacodynamic evaluation of neutralizing antibodies and virus entry inhibitors can be performed efficiently by the system,which can provide a technical platform for evaluation of neutralizing antibody of SARS-CoV-2 and screening of virus entry inhibitors,and would benefit R&D of anti-SARS-CoV-2 drugs.

Although the development of COVID-19 vaccines has been a remarkable success, the heterogeneous individual antibody generation and decline over time are unknown and still hard to predict. In this study, blood samples were collected from 163 participants who next received two doses of an inactivated COVID-19 vaccine (CoronaVac®) at a 28-day interval. Using TMT-based proteomics, we identified 1,715 serum and 7,342 peripheral blood mononuclear cells (PBMCs) proteins. We proposed two sets of potential biomarkers (seven from serum, five from PBMCs) at baseline using machine learning, and predicted the individual seropositivity 57 days after vaccination (AUC = 0.87). Based on the four PBMC's potential biomarkers, we predicted the antibody persistence until 180 days after vaccination (AUC = 0.79). Our data highlighted characteristic hematological host responses, including altered lymphocyte migration regulation, neutrophil degranulation, and humoral immune response. This study proposed potential blood-derived protein biomarkers before vaccination for predicting heterogeneous antibody generation and decline after COVID-19 vaccination, shedding light on immunization mechanisms and individual booster shot planning.
Humans ; COVID-19 Vaccines ; Leukocytes, Mononuclear ; Proteomics ; COVID-19/prevention & control* ; Vaccination ; Antibodies ; Antibodies, Viral ; Antibodies, Neutralizing

Objective:To investigate the relationship between serum vascular endothelial growth factor (VEGF), peripheral blood microRNA-126 (miR-126) and the number and distribution of cerebral microbleeds (CMBs).Methods:Consecutive patients with non-acute ischemic cerebrovascular disease admitted to the Department of Neurology, the First Affiliated Hospital of Baotou Medical College from June 2019 to June 2020 were enrolled. The clinical data were collected, 3.0 T MRI examination was performed, and susceptibility-weighted imaging was used to detect CMBs. The serum VEGF concentration was detected by enzyme-linked immunosorbent assay, and miR-126 was detected by fluorescence quantitative polymerase chain reaction. Multivariate logistic regression analysis was used to determine the independent influencing factors of CMBs. Multiple linear regression analysis was used to determine the correlation between serum VEGF concentration, miR-126 in peripheral blood and the number of CBMs. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of serum VEGF concentration and relative expression of miR-126 in peripheral blood for CMBs. Results:A total of 193 patients with non-acute ischemic cerebrovascular disease were enrolled, including 110 patients (57.0%) in the non-CMBs group, 20 (10.4%) in the strictly lobar CMBs group and 63 patients (32.6%) in non-strictly lobar CMBs group. The comparison among the three groups showed that age might be a risk factor for strictly lobar CMBs, while higher VEGF, higher cystatin C level, lower relative expression of miR-126 in peripheral blood, hypertension and previous stroke or transient ischemic attack might be the risk factors for non-strictly lobar CMBs. Multivariate logistic regression analysis showed that higher serum VEGF concentration was an independent risk factor for non-strictly lobar CMBs (odds ratio 1.186, 95% confidence interval 1.035-1.358; P=0.014), while the higher relative expression of miR-126 was an independent protective factor for non-strictly lobar CMBs (odds ratio 0.154, 95% confidence interval 0-0.269; P=0.026). Multiple linear regression analysis showed that higher serum VEGF concentration ( r=0.848, P<0.001) and the lower relative expression of miR-126 ( r=-0.043, P=0.035) significantly increased the number of CMBs. ROC curve analysis showed that the area under the curve of serum VEGF for predicting non-strictly lobar CMBs was 0.803 (95% confidence interval 0.741-0.865), the optimal cut-off value was 120.55 ng/L, the sensitivity was 70.7%, and the specificity was 75.5%. Conclusions:In patients with non-acute ischemic cerebrovascular disease, there is a significant correlation between serum VEGF concentration and the relative expression of miR-126 in peripheral blood and the number and distribution of CMBs. Serum VEGF can be used as a biomarker for predicting the presence of non-strictly lobar CMBs.

Objective    To systematically evaluate the expression of programmed cell death receptor 1 (PD-1) and programmed cell death ligand 1 (PD-L1) in esophageal squamous cell carcinoma and its relationship with prognosis. Methods    The literature from PubMed, EMbase, The Cochrane Library, Web of Science, CNKI and Wanfang data from inception to February 22, 2020 was searched by computer. Data were extracted and the quality of literature was evaluated using RevMan 5.3 software for meta-analysis. Egger's and Begg's tests were used to evaluate publication bias, and Stata 15.1 software was used for sensitivity analysis. Results     A total of 16 articles were included, and there were 3 378 patients with esophageal squamous cell carcinoma. The methodological index for nonrandomized studies (MINORS) scores were all 12 points and above. The meta-analysis results showed that the positive expression rates of PD-1 and PD-L1 in tumor cells were 37.8% (190/504) and 41.7% (1 407/3 378), respectively. The positive expression of PD-L1 in tumor immune infiltrating cells was 41.7% (412/987). The overall survival (OS) of the tumor cell with high PD-L1 expression was lower than that with low PD-LI expression (HR=1.30, 95%CI 1.01-1.69, P=0.04). The OS of the tumor immune infiltrating cell with high PD-L1 expression was significantly higher than that with low PD-LI expression (HR=0.65, 95%CI 0.53-0.80, P<0.000 1). Conclusion    PD-L1 has a high expression rate in esophageal squamous cell carcinoma and is an important factor for the prognosis of esophageal squamous cell carcinoma.

Objective: To investigate the expression of inhibitory receptor TIGIT gene in peripheral NK cells of patients with rheumatoid arthritis (RA) and its clinical significance. Methods: A case control study was conducted of 58 RA patients(30 patients with active RA disease, 28 patients with remission of RA) and 22 healthy controls (HC) in the department of rheumatology and immunology from Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine during December 2018 to June 2019, the related clinical data were collected. Flow cytometry was used to compare the expression of TIGIT gene on peripheral NK cells. The IFN-γ secretion level of cytokines in peripheral blood was detected by ELISA, and analyzed the correlations between TIGIT gene and disease activity and IFN-γ level. T-test or non-parametric test was used for comparison between the two groups, and Pearson correlation analysis was used for correlation between the two variables. Results: Compared with HC group, the number of NK cells in RA disease group was reduced, which was (13.88±4.56) ×10⁷ cells/L in RA disease group and (25.69± 2.48) ×10⁷ cells/L in HC group (t=-2.036, P=0.041). The percentage of TIGIT gene expression in peripheral blood NK cells was not statistically different between RA disease group (39.73±9.37)% and HC group (45.64±9.91)% (t=-1.241, P=0.218). However, the average fluorescence intensity (MFI) of TIGIT gene expression was decreased, which was (7.21±2.03) in RA group and (9.01±3.29) in HC group (t=-2.947, P=0.004).MFI of TIGIT gene in NK cells of the disease active subgroup was (6.72±2.01), lower than that of the disease remission subgroup (8.75±2.64), (t=-3.316, P=0.002), and MFI of TIGIT gene in NK cells of the disease active subgroup was negatively correlated with DAS28 score (r²=0.649 6, P<0.000 1).The secretion level of IFN-γ cytokine in the RA disease group was (67.13±14.84) pg/ml, higher than that in the HC group (57.21±14.23) pg/ml (t=2.757, P=0.017), and the secretion level of IFN-γ cytokine in the disease active subgroup was negatively correlated with the MFI of TIGIT gene on NK cells (r²=0.662 2, P<0.000 1). Experimental results of peripheral blood mononuclear cell stimulation in the RA disease activity subgroup showed that the secretion level of cytokines IFN-γ was reduced after stimulation compared with that before stimulation (t=11.38, P<0.000 1). Conclusion The abnormal expression of TIGIT gene on peripheral NK cells are observed in patients with RA, which correlate with disease activity and IFN-γ secretion level.

Since 2014, the United States and Europe has approved all oral, interferon free- regimens that combine with direct-acting antiviral agents. Hence, the sustained virological response rate of patients with chronic HCV genotype 1 infection has improved over 90%, and the treatment modalities has introduced a new era. These drugs, ombitasvir and dasabuvir, received customary authorization of Food and Drug Administration in 2015 and are the first combined direct-acting antiviral agents for treating HCV genotype 1 infection. It has superior application prospects in China because of its high-sustained virological response rate and safety profile. This article reviews the pharmacokinetics, drug interactions, efficacy and safety of this therapeutic regimen.

Objective To assess the impact of postnatal exposure to antibiotics on intestinal microbiome in preterm infants with 16S rDNA sequencing technology.Methods This study was conducted on 19 preterm infants admitted to the neonatal intensive care unit (NICU) at Tongji Hospital immediately after birth from September 2015 to February 2016.Two groups were set up according to the duration of antibiotic exposure (＜3 d,n=10;＞7 d,n=9).Fecal samples were collected from each infant within the first day and 2 or 3 weeks after bitrth.High-throughput sequencer (Hiseq 2500) was used for sequencing,from which information on composition and abundance of species,phylogenetic evolution and bacterial community diversity was obtained.Intergroup differences was analyzed with independent samples t-test or Fisher's exact test.Results (1) No statistically significant difference was found in general information about the infants between the two groups.(2) The intestinal flora in preterm infants was mainly composed of Lactococcus,Enterococcus and Bacillus for both groups before antibiotic treatment (36.41％,23.40％ and 14.98％).The proportions of Lactococcus and Bacillus were decreased significantly (1.73％ and 1.25％,P＜0.01) with Enterococcus becoming the predomiant genus (16.73％) after antibiotic treatment,while the relative proportions of Staphylococcus,Clostridium and Bifidobacterium were raised.(3) The Shannon index was decreased after antibiotic exposure [(2.34±0.84) vs (1.06±0.96) in ＜3 d group,and (2.64± 1.04) vs (0.35±0.36) in ＞7 d group,both P＜0.05],and the other three Alpha diversity indexes,including observed species,Chaol and PD whole tree indexes,were also decreased within each group (all P＜0.05).(4) Bacterial assemblages showed high beta diversity in both groups before the usage of antibiotics,but antibiotic therapy reduced the diversity.(5) Anoism analysis showed significant differences in the composition of intestinal flora within each group before and after antibiotic exposure (R=0.555and 0.733,both P=0.001),but no difference was found between the two groups after antibiotic exposure (R=0.060,P=0.138).Conclusions Antibiotic exposure,even short-term (＜3 d) administration,may significantly change the distribution of intestinal microbiota in preterm infants.Prolonged usage of antibiotics could have detrimental influence on intestinal flora.Therefore,for preterm babies,prescription of antibiotics should be cautious,even short-term empirical usage.

Objective To explore the efficacy and safety of daclatasvir (DCV ) combined with asunprevir (ASV) for chronic genotype 1b (GT1b) hepatitis C .Methods Twenty-nine GT1b hepatitis C patients who were treated with DCV combined ASV in Henan Provincial People′s Hospital from September 2017 to November 2017 were included .Hepatitis C virus (HCV ) RNA levels were tested before treatment ,1 week ,2 weeks ,3 weeks ,4 weeks ,8 weeks ,12 weeks and 24 weeks after treatment , and 12 weeks after the end of the treatment .The comorbidities ,combined use of drugs and adverse clinical events were registered .T test was used to compare the measurement data with normal distribution and M (P25,P75) was used for measurement data with non-normal distribution .Results A total of 29 patients with GT1b were included ,with 4 cirrhosis cases and 25 non cirrhotic cases .Seven patients had history of previous interferon and ribavirin combination treatment .There were 9 patients with comorbidity and 7 patients with combined medication . Finally , 25 patients completed a 24-week course of antiviral treatment ;3 patients were lost to follow-up ,and 1 patient withdrew after 16weeks of antiviral treatment because of a virus rebound .Of the 26 followed up patients ,25 achieved sustained virological response at 12-week (SVR12 ) , and one patient failed .And the HCV RNA NS5A resistance-associated variants (RAV) were detected in the patients with treatment failure .No severe adverse clinical events occurred in 26 patients .Conclusions DCV combined with ASV is effective and safe in the treatment of GT 1b chronic hepatitis C .However , the effect of RAV on therapeutic efficacy should be concerned during the treatment .