This study was aimed at comparing performance differences among three metagenomic sequencing platforms,MGISEQ-2000,Illumina NextSeq 2000,and Ion GeneStudio S5 Plus,to optimize the sequencing process for trace samples.The three sequencing platforms were used to perform high-throughput sequencing on DNA standards and simulated samples.Through analysis of the quality of raw data and microbial detection capabilities,systematic differences among platforms were compared.The sequencing results were optimized for trace samples by incorporation of exogenous nucleic acids during the li-brary preparation process.In terms of data output per batch and base quality,MGISEQ-2000 surpassed the other two plat-forms.Illumina NextSeq 2000 had the lowest proportion of duplicate reads,whereas Ion GeneStudio S5 Plus had the highest proportion,and significant differences were observed across platforms(P＜0.001).In sequencing uniformity,MGISEQ-2000 and Illumina NextSeq 2000 were superior to Ion GeneStudio S5 Plus.MGISEQ-2000 provided a substantial advantage in microbial detection capability(P＜0.001),but the advantage diminished with decreasing bacterial fluid concentration.Ion GeneStudio S5 Plus had the shortest duration for single-batch sequencing.Moreo-ver,for trace samples with DNA content ≤0.05 ng,the experi-mental group(with added exogenous nucleic acids)achieved a higher number of reads than the control group(without exogenous nucleic acids),with a 11.09±8.03 fold increase.In conclu-sion,the different sequencing platforms each had advantages and disadvantages,thus allowing researchers to choose the appro-priate platform according to specific needs.Furthermore,the addition of exogenous nucleic acids improved the microorganism detection efficiency,and provided better support for subsequent diagnosis and evaluation of results.

Objective To establish the fingerprints of QiangliDingxuan tablets(QLDX),determine their pharmacody-namic indexes of vasodilation and study the spectrum-effect relationships between the chemical components of QLDX and the property of vasodilation.Methods The rate of vascular relaxation was used as an index to evaluate the extent to which QLDX relaxed isolated superior mesenteric artery ring.The Grey correlation degree and partial least square regression(PLSR)were used to analyze"spectrum-effect"correlations before components with greater contribution to drug efficacy were screened out.Results There were 21 common peaks in the HPLC fingerprint of QLDX,and the similarity exceeded 0.88.A comparison with the chromatogram of the reference substance revealed 14 characteristic peaks.Vasodilative experiments showed that all the 10 batches of samples had vasodilatory effects.The correlation between the 21 chromato-graphic peaks was greater than 0.73.PLSR showed that 11 components were positively correlated with the vasodilatory effect.Six known compounds included parishin A,parishin C,5-hydroxymethylfurfural,luteolin,Linarin and ligustrazine.Conclusion The vasodilatory effect of QLDX results from the combined action of multiple components.Parishin A,parishin C,5-hydroxymethylfurfural,luteolin,Linarin and ligustrazine are positively correlated with this effect,which may be the main pharmacodynamic substance basis of vascular relaxation.

This study aims to separate and characterize self-assembled nanoparticles(SAN) from Shaoyao Gancao Decoction(SGD) and determine the content of active compounds. Further, we aimed to observe the therapeutic effect of SGD-SAN on imiquimod-induced psoriasis in mice. The separation of SGD was performed by dialysis, and the separation process was optimized by single factor experiment. The SGD-SAN isolated under the optimal process was characterized, and the content of gallic acid, albiflorin, paeoniflorin, liquiritin, isoliquiritin apioside, isoliquiritin, and glycyrrhizic acid in each part of SGD was determined by HPLC. In the animal experiment, mice were assigned into a normal group, a model group, a methotrexate group(0.001 g·kg~(-1)), and SGD, SGD sediment, SGD dialysate, and SGD-SAN groups of different doses(1, 2, and 4 g·kg~(-1)) respectively. The psoriasis grade of mice was evaluated based on the pathological changes of skin lesions, the content of inflammatory cytokines, organ index and other indicators. The results showed that SAN obtained by centrifugation at 13 000 r·min~(-1) for 30 min was stable after dialysis for 4 times, which were uniform spherical nanoparticles with the particle size of(164.43±1.34) nm, the polydispersity index of(0.28±0.05), and the Zeta potential of(-12.35±0.80) mV. The active compound content accounted for more than 70% of SGD. Compared with the model group, SAN and SGD decreased the skin lesion score, spleen index, and inflammatory cytokine levels(P<0.05 or P<0.01) and alleviated the skin thickening and infiltration of inflammatory cells. However, the sediment group and the dialysate group had no obvious effect. SGD showed a good therapeutic effect on imiquimod-induced psoriasis in mice, and SAN demonstrated the effect equivalent to SGD in a dose-dependent manner. Therefore, we conclude that the SAN formed during decocting is the main active form of SGD, which can lower the levels of inflammatory cytokines, promote the normal differentiation of keratinocytes, and reduce the infiltration of inflammatory cells in the treatment of psoriasis lesions in mice.
Mice ; Animals ; Imiquimod ; Drugs, Chinese Herbal/pharmacology* ; Glycyrrhizic Acid ; Chromatography, High Pressure Liquid/methods*

Objective: To compare the efficacy of intravascular ultrasound (IVUS) and coronary angiography guided drug eluting stent (DES) implantation for the treatment of left main coronary artery (LMCA) lesions. Methods: Randomized controlled trials (RCT) and observational studies, which compared IVUS with coronary angiography guided DES implantation for the treatment of LMCA lesions published before August 2021 were searched in PubMed, Embase and Cochrane Library databases. Baseline data, interventional procedures and endpoint events of each study were collected. The primary endpoint was major cardiovascular adverse events (MACE), and the secondary endpoints were all-cause death, cardiac death, myocardial infarction (MI), target lesion revascularization (TLR) and target vessel revascularization (TVR). The Newcastle-Ottawa Scale (NOS) and the Cochrane Collaboration Risk of Bias tool were used to evaluate the quality of the included studies. Results: Nine studies were included, including 3 RCT and 6 observational studies, with a total of 5 527 cases of LMCA. All the 6 observational studies had NOS scores≥6, and the 3 RCT had a low risk of overall bias. The results of meta-analysis showed that compared with coronary angiography guided group, MACE rate (OR=0.55, 95%CI 0.47-0.66, P<0.001), all-cause death (OR=0.56, 95%CI 0.43-0.74, P<0.001), cardiac death (OR=0.43, 95%CI 0.30-0.61, P<0.001), MI (OR=0.64, 95%CI 0.52-0.79, P<0.001), TLR (OR=0.49, 95%CI 0.28-0.86, P=0.013) and TVR (OR=0.77, 95%CI 0.60-0.98, P=0.037) were all significantly lower in the IVUS guided group. Conclusions: Compared with angiography guided, IVUS guided PCI with DES implantation in LMCA lesions could significantly reduce the risk of MACE, death, MI, TLR and TVR. IVUS is thus superior to coronary angiography for guiding PCI treatment among patients with LMCA.
Humans ; Coronary Artery Disease/complications* ; Coronary Angiography ; Drug-Eluting Stents/adverse effects* ; Treatment Outcome ; Percutaneous Coronary Intervention/methods* ; Ultrasonography, Interventional/methods* ; Risk Factors ; Myocardial Infarction/etiology*

Objective:To investigate the relationship between serum free triiodothyronine (FT 3) and the severity and prognosis of elderly patients with coronavirus disease 2019 (COVID-19). Methods:Clinical data of the elderly patients aged≥65 years old with COVID-19 who were admitted to the Ninth People′s Hospital of Wuxi from December 27, 2022 to January 18, 2023 were collected. The laboratory examinations of the non-severe and the severe/critical groups were compared. Patients were divided into group T1 (FT 3<2.51 pmol/L), group T2 (FT 3=2.51 to 2.95 pmol/L, ), and group T3 (FT 3>2.95 pmol/L) according to the FT 3 level. Independent sample t test was used for comparison between groups. Logistic regression analysis was performed to evaluate the risk factors for severe/critical disease, and to analyze the risk of severe/critical COVID-19 in elderly patients with different FT 3 levels. The receiver operator characteristic (ROC) curve was drawn to analyze the predictive value of FT 3 on the occurrence of severe/critical disease in patients. Kaplan-Meier survival curve and log-rank test were used to analyze the 30-day survival rate of elderly COVID-19 patients. Results:Among the 190 patients, the FT 3 level in the severe/critical group was (2.54±0.30) pmol/L, which was significantly lower than that in the non-severe group ((2.91±0.69) pmol/L), and the difference was statistically significant ( t=-3.42, P=0.001). Elevated serum FT 3 level was a protective factor for severe/critical disease in elderly COVID-19 patients (odds ratio ( OR)=0.139, 95% confidence interval ( CI) 0.034 to 0.577, P=0.007). There were 66 patients in the T1 group, 61 in the T2 group and 63 in the T3 group, respectively. After adjusted for sex, age, history of lung diseases, history of cardiac diseases, history of hypertension, history of diabetes mellitus, history of cerebral infarction, white blood cell count, lactate dehydrogenase, D-dimer, free thyroxine and thyrotropin, the risk of developing severe/critical disease in group T1 and group T2 were 10.982 folds and 3.695 folds of that in group T3, respectively (both P<0.05). The area under ROC curve of FT 3 of predicting severe/critical COVID-19 in the elderly patients was 0.731. The sensitivity and specificity were 0.733 and 0.659, respectively. The cut-off value was 2.53 pmol/L. The 30-day survival rate in patients with FT 3<2.53 pmol/L was significantly lower than that in patients with FT 3≥2.53 pmol/L ( χ2=13.49, P<0.001). Conclusions:The elevated level of FT 3 is a protective factor for progression to severe/critical disease in elderly patients with COVID-19. The evaluation of serum FT 3 could predict the severity and prognosis of elderly COVID-19 patients.

Silk fibroin is a natural polymer with certain water solubility, structural modification, good biocompatibility and biodegradability, which can be used as a drug delivery carrier material. As a promising drug delivery system, drug-loaded silk fibroin nanoparticles can control drug release, reduce toxicity and improve therapeutic effects. In this paper, the basic characteristics of silk fibroin, the preparation methods of drug-loaded silk fibroin nanoparticles and the application of silk fibroin in nanoparticulate drug delivery systems are reviewed, and on this basis, the further development of drug-loaded silk fibroin nanoparticles is prospected.

Objective: To evaluate the efficacy within the first 24 h post extracorporeal membrane pulmonary oxygenation (ECMO) and the impact of early efficacy on the prognosis of adult patients with fulminant myocarditis (FM). Methods: This retrospective case analysis study included hospitalized patients (age≥18 years) who were diagnosed with fulminant myocarditis from November 2016 to May 2021 in the First Affiliated Hospital of Zhengzhou University. Patients were divided into survival or non-survival groups according to treatment outcomes. The age, sex, treatments, drug use, ECMO use, clinical and laboratory data (before and 24 h after the use of ECMO) were analyzed. The change rate of clinical and laboratory data after 24 h use of ECMO was calculated to find differences between two groups. Multivariate logistic regression was used to analyze the related factors with in-hospital death and complication between the two groups. Results: A total of 38 FM patients treated with ECMO were included. There were 23 cases (60.5%) in the survival group, aged (39.6±13.7) years, and 17 (73.9%) cases were female. The total ECMO time was (134.4±71.3)h. There were 15 cases (39.5%) in non-survival group, aged (40.0±15.8) years, and there were 12(80.0%) female, the ECMO time was (120.1±72.4) h in this group. The proportion of tracheal intubation and continuous renal replacement therapy in the survivor group and dosage of norepinephrine within 24 h after ECMO implantation were significantly less than in non-survival group (all P<0.05). There was no significant difference in all efficacy related biochemical indexes between two groups before ECMO use. The levels of lactic acid, procalcitonin, creatinine, alanine aminotransferase, aspartate aminotransferase, creatine kinase-MB, cardiac troponin I and N-terminal B-type natriuretic peptide prosoma were significantly less in survival group than in non-survival group at 24 h after the use of ECMO (all P<0.05). Results of multivariate logistic regression analysis showed that the higher 24 h change rate of creatinine (OR=0.587, 95%CI 0.349-0.986, P=0.044) and creatine kinase-MB (OR=0.177, 95%CI 0.037-0.841, P=0.029) were positively correlated with reduced risk of in-hospital mortality. The central hemorrhage and acute kidney injury in survival group were less than in non-survivor group (P<0.05). Conclusions: After 24 h early use of ECMO in FM patients, the improvement of various efficacy related biochemical test indexes in the survival group was better than that in the non-survival group. Faster reduction of creatine kinase-MB and creatinine values within 24 h ECMO use is positively correlated with reduced risk of in-hospital mortality in adult patients with FM.
Adolescent ; Adult ; Extracorporeal Membrane Oxygenation/methods* ; Female ; Hospital Mortality ; Humans ; Middle Aged ; Myocarditis/therapy* ; Retrospective Studies ; Treatment Outcome ; Young Adult

AIM: To study the effect of leech extract on the expression of vascular endothelial growth factor(VEGF)in retinoblastoma(WERI-RB-1 cells)and the related molecular mechanism.METHODS: WERI-RB-1 cells cultured in vitro were divided into control group, 0.04U/mL leech extract group and 0.08U/mL leech extract group. The control group was cultured with complete medium for 48h, and the leech extract group was cultured with 0.04 and 0.08U/mL drug-containing medium for 48h, respectively. The expression level of VEGF in conditioned medium of cell culture was detected by ELISA. The mRNA levels of hypoxia-inducible factor-1a(HIF-1a)and matrix metalloproteinase-9(MMP-9)were detected by RT-PCR. The expression levels of HIF-1a, MMP-9, phosphatidylinositol 3-kinase(PI3K)and human phosphorylated AKT(p-AKT)were detected by Western Blot.RESULTS: Compared with the control group, the expression of VEGF in the conditioned medium with the concentration of 0.04U/ mL and 0.08U/mL of leech extract was decreased(P<0.05), and the inhibition rate was 32.43% and 38.92%, respectively. The expression levels of HIF-1a and MMP-9 mRNA were significantly decreased(P<0.05). The inhibition rates of HIF-1a mRNA expression of the two leech extracts were 27.64% and 24.75%, respectively. The inhibition rate of MMP-9 mRNA expression was 43.97% and 51.48%, respectively. The protein expression levels of HIF-1a, MMP-9, PI3K and p-AKT were significantly decreased compared with the control group(P<0.05). The inhibition rates of 0.04 and 0.08U/mL leech extracts on the protein expression of HIF-1a, MMP-9, PI3K and p-AKT were 55.81% and 43.85%, 39.49% and 47.23%, 33.27% and 29.83%, 52.07% and 30.21%, respectively.CONCLUSION:Leech extract may inhibit the expression of VEGF in WERI-RB-1 cells via the VEGF/PI3K/AKT pathway or HIF-1a and MMP-9 factors.

OBJECTIVE: To evaluate the diagnostic performances of salivary gland ultrasonography(SGUS)in Sjögren's syndrome(SS). METHODS: A total of 246 patients with dry mouth and/or eyes who were treated in the outpatient department and inpatient department of Rheumatology and Immunology Department of the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine from December 2019 to January 2022 were collected. All patients received SGUS examination and scored by 2019 outcome measures in rheumatology clinical trial (OMERACT)ultrasonic scoring system.Their general information, unstimulated saliva flow rate(USFR), Schirmer test and serological test results were recorded. In the study, 193 cases had lip gland biopsy. The 2016 American College of Rheumatology(ACR)/ European League Against Rheumatism(EULAR)classification criteria were adopted as the diagnostic standard of SS. χ² test was used to compare the difference of salivary gland ultrasonic scores between the two groups. The receiver operating characteristic(ROC) curve was used to evaluate the accuracy of SGUS in diagnosing SS, and the disease characteristics of SGUS positive group and negative group in the SS patients were compared. RESULTS: A total of 175 patients were SS group according to the ACR/EULAR classification, and the remaining 71 patients were non-SS group.There was no significant difference in age [(54.2±11.8) years vs. (53.4±14.9) years, P=0.705] and female (94.4% vs.93.1%, P=1.000) between SS and non-pSS groups. A total of 109 patients were SGUS positive (≥ 2 points), of whom 104 patients met the SS diagnosis and 5 patients did not meet the SS diagnosis. The positive rate of SGUS in SS group was significantly higher than that in non-SS group (59.4% vs. 7.0%, P < 0.001). The accuracy of 2019 OMERACT ultrasonic scoring system to predict ACR/EULAR classification was good, with an area under the curve of 0.762 (95%CI 0.701-0.823). The absolute agreement between the SGUS outcome and ACR-EULAR classification was 69.1%(170/246), with a sensiti-vity of 59.4%(104/175), specificity of 93%(66/71), positive predictive value of 95.4%(104/109) and negative predictive value of 48.2% (66/137). A total of 81 patients were positive SGUS combined with anti-SSA antibody, 100% (81/81) fulfilled the ACR-EULAR criteria, 85 patients were negative SGUS and anti SSA antibody, and 60 patients(70.6%, 60/85) did not fulfil the ACR-EULAR criteria. SGUS positive group had higher antinuclear antibody(ANA) positive rate(83.1% vs. 98.1%, P < 0.001) in the patients with SS. CONCLUSION The OMERACT ultrasonic scoring system has high diagnostic value in SS. The combination of SGUS and anti-SSA antibody can improve the diagnostic value.
Humans ; Female ; Adult ; Middle Aged ; Aged ; Sjogren's Syndrome/diagnostic imaging* ; Sensitivity and Specificity ; China ; Salivary Glands/pathology* ; Ultrasonography/methods*

The gut microbiota takes part in many in vivo important physiological activities of host, such as the substance metabolism and energy exchange, etc. The interaction between the host and the intestinal microorganisms has attracted scholars' attention. Flavonoids are a group of polyphenol compounds widely found in natural plants, with the bioactive effect of regulation of glucose and lipid metabolism, anti-inflammation. However, their low bioavailability cause difficulty to clarify the effective substances and the mechanism of flavonoids. Apart from the metabolic effects of liver on flavonoids, recent studies have shown that the gut microbiota can interact with flavonoids. On the one hand, flavonoids can be metabolized by gut microbiota and subsequent metabolites can produce pharmacological activities different from the parent components. On the other hand, flavonoids and their metabolites can in turn regulate the composition and physiological activities of the intestinal flora, which seems to provide a new insight for the research on the effective substances of flavonoids. In this review, we introduced the metabolic characteristics of flavonoids under the actions of intestinal bacteria, and the regulation effects of flavonoids on gut microbiota was also summarized. Meanwhile, the therapeutic effect of flavonoids under the action of intestinal bacteria was discussed.