The PA-PB1 interface of the influenza polymerase is an attractive site for antiviral drug design. In this study, we designed and synthesized a mini-library of indazole-containing compounds based on rational structure-based design to target the PB1-binding interface on PA. Biological evaluation of these compounds through a viral yield reduction assay revealed that compounds 27 and 31 both had a low micromolar range of the half maximal effective concentration (EC₅₀) values against A/WSN/33 (H1N1) (8.03 μmol/L for 27; 14.6 μmol/L for 31), while the most potent candidate 24 had an EC₅₀ value of 690 nM. Compound 24 was effective against different influenza strains including a pandemic H1N1 strain and an influenza B strain. Mechanistic studies confirmed that compound 24 bound PA with a K _d which equals to 1.88 μmol/L and disrupted the binding of PB1 to PA. The compound also decreased the lung viral titre in mice. In summary, we have identified a potent anti-influenza candidate with potency comparable to existing drugs and is effective against different viral strains. The therapeutic options for influenza infection have been limited by the occurrence of antiviral resistance, owing to the high mutation rate of viral proteins targeted by available drugs. To alleviate the public health burden of this issue, novel anti-influenza drugs are desired. In this study, we present our discovery of a novel class of indazole-containing compounds which exhibited favourable potency against both influenza A and B viruses. The EC₅₀ of the most potent compounds were within low micromolar to nanomolar concentrations. Furthermore, we show that the mouse lung viral titre decreased due to treatment with compound 24. Thus our findings identify promising candidates for further development of anti-influenza drugs suitable for clinical use.

Background Staff in biosafety laboratories (BSL) are more likely to experience occupational burnout and other psychological issues due to their unique working environment and high job demands. However, current research in this field tends to focus on overall analyses, overlooking the internal differences within this group. Objective To explore latent profiles of occupational burnout among BSL workers and their influencing factors, providing a reference for targeted burnout interventions. Methods In 2022, cluster random sampling was used to select 8023 BSL workers in Xinjiang. Data were collected via a questionnaire survey, including a self-designed general information questionnaire, the Maslach Burnout Inventory-General Survey (MBI-GS), and the Effort-Reward Imbalance (ERI) scale, to analyze demographic characteristics, occupational burnout, and job stress status. After data cleaning, latent profile analysis (LPA) was applied to identify burnout profiles, followed by K-means clustering for verification. Logistic regression was used to analyze influencing factors with burnout profiles as the dependent variable. Results After excluding invalid questionnaires, 7924 valid questionnaires were retained, with an recovery rate of 98.77%. The LPA results indicated that occupational burnout among BSL staff was categorized into three latent profiles: a low burnout profile (2208 individuals, 27.86%), a moderate burnout profile (4636 individuals, 58.51%), and a high burnout profile (1080 individuals, 13.63%). A similar grouping was derived from K-means clustering analysis (28.71%, 53.37%, and 17.92%), and a Sankey diagram demonstrated significant overlap between the two classification results. Statistically significant differences were observed across the three latent profiles in all three dimensions of burnout: emotional exhaustion, cynicism, and reduced personal accomplishment. The final logistic regression model revealed that the influencing factors exhibited characteristics of partial overlap and partial differentiation. Among the common influencing factors, the shared protective factors included: age ≥50 years (OR_{moderate burnout}=0.42, OR_{high burnout}=0.45), monthly income of 4000-6000 RMB (OR_{moderate burnout}=0.83, OR_{high burnout}=0.71), good sleep quality (OR_{moderate burnout}=0.70, OR_{high burnout}=0.13) and very good sleep quality (OR_{moderate burnout}=0.51, OR_{high burnout}=0.12), and engaging in physical exercise ≥5 times per week (OR_{moderate burnout}=0.74, OR_{high burnout}=0.48). The protective effects were more pronounced for the high burnout profile. The common risk factors included: 21–30 years of service (OR_{moderate burnout}=1.38, OR_{high burnout}=3.00), ≥31 years of service (OR_{moderate burnout}=1.47, OR_{high burnout}=3.39), working in "other positions" (OR_{moderate burnout}=1.46, OR_{high burnout}=1.56), and occupational stress (ERI>1) (OR_{moderate burnout}=2.62, OR_{high burnout}=3.25), with the risk effects being more significant for the high burnout profile. Regarding differentiated influencing factors, protective factors exclusively associated with the moderate burnout profile included having an associate degree (OR=0.70) and engaging in physical exercise 3-4 times per week (OR=0.74). For the high burnout profile, exclusive protective factors included age 40-50 years (OR=0.56), while exclusive risk factors included having a senior professional title (OR=1.85), holding a managerial role (OR=1.78), a high number of night shifts per month (OR=2.66), and a very high number of night shifts per month (OR=3.93). Conclusion BSL workers exhibit three distinct latent burnout profiles. Occupational burnout requires greater attention, and targeted prevention and intervention strategies should be developed based on worker subtypes.

From October 2021 to February 2023, we retrospectively analyzed the clinical and laboratory data of six patients (three male and three female, median age: 54 years, age range: 29-73 years) with mast cell leukemia (MCL) diagnosed in the First Affiliated Hospital of Soochow University (The Mastocytosis Collaborative Network of China). All patients had acute MCL, with at least one C-finding present. The main clinical presentations were hypoalbuminemia ( n=4), fatigue ( n=3), fever ( n=2), abdominal discomfort ( n=2), osteolytic lesions ( n=2), dizziness ( n=1), skin flushing ( n=1), and weight loss ( n=1). Splenomegaly and lymphadenopathy were noted in six and three patients, respectively. Six patients were strongly positive for CD117, five were positive for CD30 and CD25, and four were positive for CD2. Four patients had a normal karyotype and two patients had an abnormal karyotype. Gene mutations were detected in 4/6 cases. The median serum tryptase level was 24.9 (range: 20.1-171.9) μg/L. Two patients were treated with venetoclax and azacitidine for induction (one patient achieved partial remission by combination with afatinib, while there was no remission after combination with dasatinib in the other patient). Two patients did not achieve complete remission despite treatment with cladribine and imatinib, respectively. One patient treated with interferon combined with glucocorticoids was lost to follow-up, and one patient abandoned treatment. The follow-up time ranged from 1.1 to 21.7 months. Three patients died and two survived. Overall, MCL is a rare subtype of systemic mastocytosis with heterogeneous clinical course, and these patients have poor outcome. A better understanding of the clinical characteristics, treatment, and prognosis of MCL is urgently needed.

Objective:To investigate the role of complement C3a receptor in the diabetic nephropathy pathogenesis of db/db mice, and to provide a new target for prevention and treatment of diabetic nephropathy.Methods:Twelve 8-week-old male mice with type 2 diabetes mellitus (db/db mice) and 6 wild-type (db/m) mice were reared in the special pathogen free environment. The mice were grouped into db/m group, db/db group and C3a receptor antagonist group, with 6 mice in each group. db/db model mice were intraperitoneally injected with C3a receptor antagonist (SB290157, 10 mg/kg) once every two days for 8 weeks in C3a receptor antagonist group. Blood and urine samples were collected, and body weight of mice, fasting blood glucose, serum creatinine, blood urea nitrogen, urinary microalbumin/urinary creatinine and urinary N-acetyl-β- D-glucosaminidase (NAG) were detected. Renal tissues were collected, and HE, PAS and Masson stainings were used to observe the pathological changes. Immunohistochemistry, immunofluorescence and Western blotting were used to detect the protein expression levels of C3 and C3a receptor. Western blotting was used to analyze the protein expression levels of kidney injury molecule-1 (Kim-1), α-smooth muscle actin (α-SMA), zonula occluden-1 (ZO-1), vimentin and E-cadherin in renal tissues. Immunofluorescence was used to analyze the protein expression levels and distribution of α-SMA, ZO-1 and Kim-1, and immunohistochemistry was used to analyze the protein expression levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α). TUNEL assay was used to detect apoptotic cells in renal tissues. Results:Compared with db/m group, body weight, fasting blood glucose, urinary microalbumin/urinary creatinine and urinary NAG in db/db group were significantly higher, while these indicators in C3a receptor antagonist group were slightly lower than those in db/db group (all P<0.01). There were no significant differences in serum creatinine and blood urea nitrogen among the three groups (all P>0.01). Compared with db/m group, db/db group had glomerular hypertrophy, necrosis and exfoliation of renal tubular epithelial cells, and dilation of renal tubules, and C3 and C3a receptor protein expression levels were higher (both P<0.01). Compared with db/db group, C3a receptor antagonist group had less glomerular lesions, mild necrosis of renal tubular epithelial cells and less tubular dilation. Compared with db/m group, the protein expression levels of Kim-1, IL-1 and TNF-α in kidney tissues of db/db group were significantly higher, while Kim-1, IL-1 and TNF-α in C3a receptor antagonist group were significantly lower than those in db/db group (all P<0.01). Compared with db/m group, the protein expression levels of α-SMA and vimentin of renal tubular epithelial cells in db/db group were significantly higher, while the protein expression levels of ZO-1 and E-cadherin were significantly lower (all P<0.01). Compared with db/db group, the protein expression levels of α-SMA and vimentin of renal tubular epithelial cells in C3a receptor antagonist group were significantly lower, and the protein expression levels of ZO-1 and E-cadherin were significantly higher (all P<0.01). Compared with db/m group, the number of apoptotic cells of kidney tissues in db/db group was increased, while the number of apoptotic cells in C3a receptor antagonist group was reduced compared with db/db group. Conclusions:The expression levels of C3 and C3a receptor of kidney tissues in db/db mice are significantly increased. Antagonistic C3a receptor can reduce the body weight, blood glucose, urinary microalbumin/urinary creatinine and urinary NAG, alleviate renal pathological injury, inhibit renal tissue inflammation, apoptosis and renal tubule epithelial-mesenchymal transition in db/db mice.

Objective:To study the effects of Jianpi Qinghua Decoction on hepatocyte apoptosis under non-alcoholic fatty liver disease (NAFLD), and to discuss its mechanism.Methods:Totally 29 C57 mice were randomly selected and fed a 60% high fat diet for 16 weeks, while the remaining 6 mice were given regular feed as the normal group. After successful modeling, 12 mice with larger body weight were divided into TCM group and model group using a random number table method, while continuing to receive high-fat feed. The TCM group was orally administered Jianpi Qinghua Decoction 20.961 g/kg. The normal group and model group were orally administered an equal volume of distilled water once a day, with continuous intervention for 4 weeks. The levels of GOT and GPT in blood were detected by ELISA, the deposition of triglyceride in liver was detected by oil red O, and the apoptosis of liver cells was detected by TUNEL fluorescence staining. Western blot was used to detect the expressions of cleaved Caspase-3, Caspase-3, Bax, Bcl-2, JNK and p-JNK.Results:Compared with the model group, the body weight and GPT in the TCM group significantly decreased ( P<0.05), TG deposition was significantly reduced, apoptosis range of liver cells was significantly reduced, and cleaved Caspase-3/Caspase-3, p-JNK/JNK and the expression of Bax significantly decreased ( P<0.05). The expression of Bcl-2 increased ( P<0.05). Conclusion:Jianpi Qinghua Decoction can inhibit JNK protein phosphorylation and effectively reduce liver cell apoptosis in NAFLD mice, which may delay the progression of NAFLD towards cirrhosis and liver cancer.

Objective:This multicenter clinical evaluation analyzed the clinical performance of five fast nucleic acid detection systems for 2019-nCoV.Methods:Clinical performance of the five fast nucleic acid detection reagents approved in China was evaluated in the present study. Fifty-seven throat swabs samples from COVID-19 patients and fifteen throat swabs samples from healthy people were collected from the First Affiliated Hospital of Zhejiang University school of Medicine, Tongji Hospital of Tongji Medical College of HUST, and National Institute of Viral Disease Control and Prevention of CDC to evaluate the positive coincidence rate, negative coincidence rate, total coincidence rate, the detection time and retest rate as well as the relation between positive intensity and positive coincidence rate of the five fast nucleic acid detection systems in November 2020.Results:The positive coincidence rates of the five kits were 92.59% (50/54), 83.64% (46/55), 98.25% (56/57), 94.44% (51/54) and 98.18% (54/55); and the negative coincidence rates were 93.33% (14/15), 93.33% (14/15), 86.67% (13/15), 100% (14/14) and 93.33% (14/15); and the total coincidence rates were 92.75% (64/69), 85.71% (60/70), 95.83% (69/72), 94.20% (65/69) and 97.14% (68/70), respectively. The positive coincidence rate of the five kits reached 100% for the strong-positive (90/90) and medium-positive samples (84/84), but only 82.18% (83/101) for weak-positive samples (cycle threshold value>33), and the retest rate of two kits were 15.28% (11/72) and 12.50% (9/72), which were both higher than 10%. Total time from sample extraction to amplification was between 32.33-65.33 minutes for these five kits.Conclusion:The five fast nucleic acid detection reagents have good performance and can be used as a supplement to routine nucleic acid detection reagents.

Objective To establish an analysis method for quantitatively determining phenolic character-istic components and correlation analyzing their quality representation of specific chromatograms in Shaji (Seabuckthorn Fruit,Hippophae rhamnoides),and to review the quality of Shaji effectively and accurate-ly by applying association analysis-reviewing mode.Methods HPLC-PDA method was used to quantita-tively determine the content of phenolic characteristic components in 11 batches of Shaji(protocatechuic acid,ellagic acid,narcissin,quercetin,isorhamnetin),and to establish phenolic specific chromatograms of Shaji.The quality of 11 batches of Shaji was characterized based on teasing characteristic peaks and chemical types.The quantity of 11 batches of Shaji was characterized based on quantity and peak areas of protocatechuic acid, ellagic acid, narcissin, quercetin, isorhamnetin, phenolic acids(represented by protocatechuic acid)and flavonoids(represented by narcissin)in the specific chromatograms.The characterized results of quality and quantity of 11 batches of Shaji were given association analysis based on baseline material of Shaji.Results The characteristic components of protocatechuic acid, ellagic acid,narcissin,quercetin,isorhamnetin all had good linear correlation,and the results of methodological investigation were in accordance with the quantitative determination requirements.Taken batch 7 of Shaji as baseline material, there were totally 18 characteristic peaks in phenolic specific chromatograms of Shaji,including 3 peaks of phenolic acids and 15 peaks of flavonoids, and all 18 characteristic peaks appeared in the chromatograms of 11 batches of Shaji.The quantity of characteristic components were higher in batch 8, 7, 10, 4, 5, 11 and 1 after analyzed and reviewed by using association analysis-reviewing mode.The relevance of batch 6, 2, 3, 1 and 11 was the highest with baseline material of Shaji.Comprehensive reviewing showed that the excellent extents of batch 1,11,8,6 and 7 were prior. Conclusion The quantitative determination method of phenolic characteristic components in Shaji estab-lished in this study is easy and accurate.The association analysis-reviewing mode for quality characteriza-tion of phenolic specific chromatograms can be used for analyzing the quality and application validity of Shaji and reviewing quality of Shaji effectively and accurately.

In recent years, the number of ICU survivor is ever-growing with the increase of cure rate. The survivors′ outcome has aroused more and more attentions from medical personnel. ICU survivors tend to experience lasting physical, psychological and cognitive injuries, the post-traumatic stress disorder is one kind of significant psychological injury associated with patients′ critical experience. This review aims to summarize relevant literatures, introduces the prevalence, risk factors, complications, and interventions of ICU survivors′ post-traumatic stress disorder symptom, in order to prevent ICU survivors from post-ICU psychological injury and improve their long-term outcome.

This study was aimed to develop an HPLC method for simultaneous determination of five active components in Baihu and Guizhi decoction.Simultaneous determination of mangiferin,neomangiferin,glycyrrhizic acid,liquiritin,and cinnamic acid in Baihu and Guizhi decoction were conducted by HPLC-PDA under multiple UV wavelengths.An Waters xBridge BEH C18 Column (4.6 mm × 250 μm,5 μm) was used.The mobile phase was acetonitrile-0.01％ formic acid solution.The flow rate was 1 mL·min-1.The column temperature was kept at 24℃.The detection wavelength was set at 255 nm and 280 nm.The results showed that the linear range of mangiferin,neomangiferin,glycyrrhizic acid,liquiritin,and cinnamic acid was 66-1 976 μg (r =0.999 3),70-3487 μg (r =0.999 7),30-913 μg (r =0.999 5),35-1 734 μg (r =0.999 5),0.6-187 μg (r =0.999 8) in 70 min,respectively.The average recovery was between 95.29％ and 100.17％.It was concluded that the method was convenient,stable,reliable and accurate for simultaneous determination of the five components in Baihu and Guizhi decoction.It provided an evaluation method for the quality control of Baihu and Guizhi decoction and the research o its granules.

Objective To optimize the extraction process of Radix Paeoniae Alba in Baijin Capsule by orthogonal experiment.Methods The study employed the extraction rate of paeoniflorin and total glucosides of paeony as evaluation indexes. The orthogonal design was used to investigate effects of solvent volume, extraction time and extraction frequency on extraction results.Results The optimal extracting condition was extracted 3 times, with 14 fold 70% alcohol, 1.5 h for each time. Conclusion The method is simple and steady, which will provide instruction and reference to the production of Baijin Capsule.