Qingzao Jiufeitang is a famous classical formula for treating lung injury caused by warm and dryness， included in the Catalogue of Ancient Famous Classical Formulas（The First Batch）. By systematically organizing ancient and modern literature on this formula， this study analyzed and verified the origin， medicinal composition， original plants and processing， dosage and decoction method， efficacy and application of this formula. According to the research， Qingzao Jiufeitang was first recorded in Yimen Falyu in the Qing dynasty， and its creation was mainly inspired by the Ming dynasty physician MIAO Xiyong's idea of the moisturizing drugs with sweet flavour and cold nature. Based on the 2020 edition of the Pharmacopoeia of the People's Republic of China（hereinafter referred to as the Chinese Pharmacopoeia） and the textual research results of modern scholars on traditional Chinese herbal medicines， the botanical sources and processing methods of the herbs in this formula are basically clarified. Among them， Mori Folium， Gypsum Fibrosum， Ginseng Radix et Rhizoma， Sesami Semen Nigrum， Asini Corii Colla， Ophiopogonis Radix and Eriobotryae Folium are consistent with the 2020 edition of the Chinese Pharmacopoeia. The primary source of Glycyrrhizae Radix et Rhizoma is the dried roots and rhizomes of Glycyrrhiza uralensis， family Leguminosae， while the primary source of Armeniacae Semen Amarum is the dried mature seeds of Prunus armeniaca， family Rosaceae. It is recommended to use Gypsum Ustum， stir-fried Sesami Semen Nigrum， stir-fried Armeniacae Semen Amarum， Asini Corii Colla bead， and honey-fried Eriobotryae Folium， and the rest of the raw products. According to the conversion of ancient and modern doses， the recommended dosages are 11.19 g for Mori Folium， 9.33 g for Gypsum Fibrosum， 3.73 g for Glycyrrhizae Radix et Rhizoma， 2.61 g for Ginseng Radix et Rhizoma， 3.73 g for Sesami Semen Nigrum， 4.48 g for Ophiopogonis Radix， 2.61 g for Armeniacae Semen Amarum， 3.73 g for Eriobotryae Folium. The decoction method is to add 300 mL of water， decoct it down to 180 mL， remove the residue， and then add 2.98 g of Asini Corii Colla into the decoction. Take it warm after meals， two to three times a day. Qingzao Jiufeitang has the effects of clearing dryness and moistening the lungs， nourishing Yin and invigorating Qi. In ancient times， it was mainly used to treat stagnation and depression of various Qi， as well as paralysis， asthma and vomiting. In modern clinical practice， it is mostly used to treat diseases in respiratory system， otolaryngology， skin system and digestive system caused by warm-dry impairing lung， deficiency of both Qi and Yin. The above research results can provide a reference for the later development of Qingzao Jiufeitang.

AIM:To evaluate the therapeutic effect of Shen-Xiankang formula on renal interstitial fibrosis in-duced by unilateral ureteral obstruction(UUO)in mice and to elucidate its underlying mechanisms.METHODS:Thirty-two C57BL/6 mice were randomly divided into sham group,UUO model group,and Shen-Xiankang formula intervention groups receiving either a low dose(150 mg·kg-1·d-1)or a high dose(450 mg·kg-1·d-1),with 8 mice in each group.All mice except those in sham group underwent UUO to establish chronic kidney disease(CKD)model.After modeling,cor-responding doses of Shen-Xiankang or an equivalent volume of saline were administered daily for 7 d.Upon completion of treatment,renal tissues were collected for hematoxylin-eosin(HE)and Masson staining to assess tissue damage and fibro-sis.Immunohistochemistry and Western blot analyses were used to detect the markers of fibrosis,oxidative stress,and fer-roptosis.The effect of Shen-Xiankang formula on the interaction between activating transcription factor 3(ATF3)and Smad3 was assessed using co-immunoprecipitation(Co-IP).RESULTS:The untreated UUO model group exhibited nota-ble pathological changes such as expanded renal tubules and collagen deposition.Shen-Xiankang treatment significantly alleviated these changes(P<0.05).It markedly reduced Smad3 phosphorylation,ATF3,4-hydroxynonenal(4-HNE),and NADPH oxidase 4(NOX4)aberrant expression,while increasing glutathione peroxidase 4(GPX4)and solute carrier family 7 member 11(SLC7A11)expression.Co-IP results indicated a significant modulation of the ATF3-Smad3 interac-tion by Shen-Xiankang.CONCLUSION:Shen-Xiankang formula effectively mitigates UUO-induced renal interstitial fi-brosis in mice.The mechanism may involve modulating the ATF3/Smad3 interaction,which in turn attenuates oxidative stress and ferroptosis,consequently leading to the amelioration of renal fibrosis.These findings provide important insights for further research and clinical application of Shen-Xiankang formula.

Objective:To analyze the diagnostic performance of contrast-enhanced ultrasound (CEUS) in the diagnosis of papillary thyroid carcinoma (PTC) and related parameters in the diagnosis of cervical lymph node metastasis.Methods:The clinical data of 130 patients who underwent ultrasonography in Ningbo Hospital of Traditional Chinese Medicine from Jan. 2019 to Jan. 2022 were retrospectively analyzed. All patients received contrast-enhanced ultrasonography and postoperative pathological examination. According to the pathological diagnosis of benign and malignant tumors, they were divided into PTC group and benign nodule group. In the PTC group, there were 46 males and 40 females, aging 51.79±5.01 years old, ranging from 32 to 63 years old; In the nodule group, there were 25 males and 19 females, aging 52.05±4.89 years old, ranging from 33 to 64 years old. According to the presence or absence of cervical LNM, they were divided into a metastasis group of 31 cases, 17 males and 14 females; age (51.69±6.14 years), ranging from 36 to 63 years; 55 cases in the non-transfer group, 29 males and 26 females, aging (51.75±6.18) years, ranging from 36 to 62 years. Comparative analysis of different nodule properties, presence or absence of LNM, different lesion diameters in imaging manifestations and time-intensity curve (time-intensity curve, TIC) parameters differences, measurement data between groups were conducted by independent sample t test, count data between groups were compared by χ2. The receiver operating characteristic curve (receiver operating characteristic, ROC) was drawn to evaluate the diagnostic performance of TIC parameters for cervical LNM. Results:The proportion of PTC nodules with low enhancement, irregular enhancement, heterogeneous enhancement, unclear lesion boundary, and perfusion defect (72.09%, 87.21%, 88.64%, 69.77%, 70.93%) was significantly higher than that of benign nodules (38.64%, 11.36%) %, 27.27%, 77.27%, 27.27%) ( χ2=13.67, 70.75, 49.69, 25.92, 18.24, P<0.05) ; PTC nodule peak intensity (peak intensity, PI), TIC area under the curve (area under curve, AUC) was significantly lower than that of benign nodules (14.86±2.11dB vs 23.94±3.51dB, 985.14±105.31dB·s vs 1621.14±182.61dB·s) ; time to peak (TTP) was significantly higher than that of benign nodules ( 44.82±5.01s vs 36.95±4.18s) ( t=18.39, 21.36, 8.94, P<0.05) ; there was no significant difference in mean transit time (MTT) ( P>0.05) ; AUC was significantly higher than that of the non-metastatic group (16.86±2.09) dB vs (13.73±1.42) dB, (1163.54±131.41) dB·s vs (884.59±93.25) dB·s ( t=8.25, 11.46, P<0.05) ; The PI and AUC of PTC patients with lesion diameter ≤1.5 cm were significantly lower than those of patients with lesion diameter > 1.5 cm (11.56±1.94) dB vs (15.93±2.46) dB, (876.97±100.21) dB·s vs (1020.09±125.41) dB·s ( t=8.39, 5.34, P< 0.05), there was no significant difference in terms of TTP or MTT ( P>0.05) ; the AUC of PI in the diagnosis of cervical lymph node metastasis in PTC patients was 0.888 (95% CI: 0.807-0.969), the sensitivity was 90.91%, and the specificity was 77.42%; The AUC for the diagnosis of cervical lymph node metastasis in PTC patients was 0.972 (95% CI: 0.943-1.000), with a sensitivity of 87.10% and a specificity of 96.36%. Conclusion:The CEUS manifestations of PTC nodules are mostly irregular and heterogeneous low-enhancement, and the TIC-related parameters of PTC nodules and benign nodules are significantly different, and TIC-related parameters have good diagnostic efficiency for patients with cervical lymph node metastasis.

Objective: To examine the effects of bisphenol A (BPA), bisphenol S ( BPS ), bisphenol F ( BPF ) and bisphenol AF ( BPAF ) on the proliferation and oxidative stress of BRL 3A rat liver cells, and to preliminarily evaluate their mutagenicities. Methods: In vitro cultured BRL 3A rat liver cells were treated with BPA, BPS, BPF and BPAF at concentrations of 0, 5, 10, 25, 50, 100, 150 and 200 μmol/L for 48 h, respectively. Then, the cell viability was determined using the CCK-8 assay, and the half maximal inhibitory concentration ( IC50 ) was calculated. The minimum inhibitory concentration for BRL 3A cell proliferation was screened, and the intracellular reactive oxygen species ( ROS ) was measured in BRL 3A cells using the 2',7'-dichlorodihydrofluorescein diacetate ( DCFH-DA ) assay. In addition, the effects of BPA, BPS, BPF and BPAF at concentrations of 1 000, 200, 40, 8 and 1.6 μg/plate on the mutant colonies of histidine-deficient Salmonella typhimurium ( TA1535, TA97a, TA98, TA100 and TA102 ) were tested using the Ames test. Results: Treatment with BPA and BPF at concentrations of 100 to 200 μmol/L and with BPAF at concentrations of 25 to 200 μmol/L inhibited BRL 3A cell survival at a concentration-dependent manner, while exposure to BPS at concentrations of 5 to 200 μmol/L resulted in no changes in BRL 3A cell survival. The IC50 values of BPA, BPS, BPF and BPAF were 131.7, >200, 187.5 and 21.6 μmol/L against BRL 3A cells, respectively. Treatment with BPS at 100 μmol/L or BPAF at 25 μmol/L caused no significant changes in the ROS level; however, exposure to BPA at 100 μmol/L and BPF at 100 μmol/L significantly increased the ROS level. Ames test showed that BPA, BPS, BPF and BPAF did not induce mutagenicity in TA1535, TA97a, TA98, TA100 or TA102 strains. Conclusions BPAF shows the highest cytotoxicity to BRL 3A cells, and low-concentration exposure to BPS has few effects on BRL 3A cells. The cytotoxicity of bisphenols against BRL 3A cells may be associated with the induction of oxidative stress. None of the four bisphenols show mutagenic effects under the present experimental conditions.

Objective:To establish a quality evaluation method for the simultaneous determination of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin in Danqi Xinmaikang boiled powders and pieces.Methods:Quantitative analysis of multi-components was performed to determine contents of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin with Calycosin-7-O-β-D-Glucopyranoside as the reference substance by single-maker (QAMS). The chromatogram conditions were established, with C18 column as solid phase, acetonitrile-water as flowing phase, 268 nm as detecting wavelength, 1.0 ml/min as flowing rate, 30 ℃ as column temperature, and 10 μl as injection volume.Results:The relative correction factor between Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin was 1.14. There was no significant difference of measured values between the external standard method and QAMS ( P>0.05). With Calycosin-7-O-β-D-Glucopyranoside retention time of 1.00, the relative retention time of Lobetyolin was 1.51 and RSD was less than 5%. Conclusion:It is feasible and accurate to evaluate the quality of Danqi Xinmaikang boiled powders and pieces by QAMS.

Objective:To study the effect of thioredoxin domain containing protein 5 (TXNDC5)-peroxiredoxin 2 (Prx2) on the drug resistance of prostate cancer cells.Methods:Prostate cancer PC3 cells were cultured in vitro, treated with the chemotherapy drug cyclophosphamide (5, 10, 15 μmol/L) for 24 hours, and PC3 cells without any treatment was served as the control group. The expression levels of TXNDC5 in PC3 cells were detected by real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting. PC3 cells with TXNDC5 knocking down were exposed by cyclophosphamide and CCK-8 was used to detect the cell viability of siTXNDC5 group and siNC group. The content of reactive oxygen free radicals was determined by reactive oxygen detection kit. PC3 cells and its parental cyclophosphamide-resistant ones with TXNDC5 knocking down were treated by 10 μmol/L cyclophosphamide and subjected for CCK8 assay. The expression of Prx2 in PC3 cells was detected by Western blotting after TXNDC5 was silenced. Prx2 expression was silenced in PC3 cells overexpressing TXNDC5, and cell viability and reactive oxygen free radical content were detected in Vec-Ctrl group, pcTXNDC5 group, siNC group, siPrx2 group and pcTXNDC5+ siPrx2 group. Results:Compared with the control group, cyclophosphamide treatment significantly increased the expression of TXNDC5 at mRNA and protein levels in PC3 cells. After PC3 cells were treated with cyclophosphamide (10, 15 μmol/L) for 12 h, compared with the siNC group, the cell viability in the siTXNDC5 group was significantly suppressed (0.44±0.08 vs. 0.74±0.10, t=3.647, P=0.031; 0.30±0.04 vs. 0.53±0.06, t=6.115, P=0.006). When PC3 cells were treated with 10 μmol/L cyclophosphamide for 6 and 12 h, compared with the siNC group, the production of reactive oxygen free radicals in the siTXNDC5 group was significantly increased (2.68±0.19 vs. 1.58±0.26, t=-6.027, P=0.005; 4.56±0.37 vs. 2.73±0.26, t=-6.995, P=0.003). When PC3 cells and its cyclophosphamide-resistant ones were treated with 10 μmol/L cyclophosphamide for 12 h, compared with the siNC group, the cell viability was significantly inhibited in the siTXNDC5 group. Western blotting analysis showed that the expression of Prx2 was significantly reduced when TXNDC5 was silenced. Silencing Prx2 could significantly attenuate the increase of cell viability and the decrease of reactive oxygen content resulting from TXNDC5 overexpression. PC3 cells were treated with 10 μmol/L cyclophosphamide for 12 h, and the cell viabilities of the Vec-Ctrl group, pcTXNDC5 group, siNC group, siPrx2 group and pcTXNDC5+ siPrx2 group were 0.52±0.07, 0.69±0.03, 0.56±0.05, 0.43±0.05, 0.58±0.07, respectively, and there was a statistically significant difference ( F=8.868, P=0.003). Furthermore, the cell viability in the pcTXNDC5+ siPrx2 group decreased significantly when compared to that of the pcTXNDC5 group ( P=0.045). The contents of reactive oxygen free radicals in the above 5 groups were 3.26±0.46, 2.09±0.49, 3.16±0.38, 4.62±0.26, 2.87±0.36, respectively, and there was a statistically significant difference ( F=16.037, P<0.001). The content of reactive oxygen radicals in the pcTXNDC5+ siPrx2 group was higher than that of the pcTXNDC5 group ( P=0.036). Conclusion:TXNDC5 can reduce the level of reactive oxygen free radicals in prostate cancer cells by regulating the expression of Prx2, so as to promote the drug resistance of prostate cancer cells.

Objective:To understand the occupational health status of a lead-acid battery enterprise in Jiangsu Province, to observe the results of blood lead and bone mineral density (BMD) of the workers exposed to occupational lead, and to explore the effect of occupational lead exposure on BMD, so as to provide basis for the prevention and treatment of occupational lead poisoning and osteoporosis.Methods:An occupational health survey was conducted in a lead-acid battery enterprise in Jiangsu Province in January 2019. Basic information and occupational health examination results of 402 persons exposed to occupational lead were collected, and BMD was measured. Spearman rank correlation test was used to analyze the relationship between blood lead and BMD, and logistic regression analysis was used to analyze the influencing factors of BMD.Results:The blood lead level M ( P25, P75) of 402 occupational lead exposure workers was 220.5 (118.0, 307.0) μg/L, 46 workers (11.4%) had blood lead value ≥400 μg/L, and 5 workers (1.2%) ≥600 μg/L. 124 workers (30.8%) had abnormal BMD. The concentrations of lead dust and lead smoke in the workplace were <0.004-0.027 and <0.021-0.045 mg/m 3, respectively. The positions exceeding the standard point were mainly concentrated in the casting and welding group (44.4%, 4/9) of lead smoke positions. There was a statistically significant difference in the overall distribution of blood lead levels among lead exposure workers with different BMD levels, and there was a positive correlation between blood lead and BMD ( P<0.01) . The results of univariate analysis showed that there were statistically significant differences in the distribution of abnormal BMD among workers exposed to different genders, positions and blood lead levels ( P<0.01) . The results of multivariate logistic regression analysis showed that the risk of abnormal BMD in male workers was 5.069 times of that in female worker (95% CI: 2.906-8.840, P<0.01) . Conclusion:Occupational lead exposure personnel have a high blood lead level and a high abnormal BMD rate. Exposure to lead working environment is an influencing factor for the abnormal BMD of workers, so enterprise managers should pay attention to health protection, occupational health monitoring and supervision of working environment of front-line workers.

Objective:To understand the occupational health status of a lead-acid battery enterprise in Jiangsu Province, to observe the results of blood lead and bone mineral density (BMD) of the workers exposed to occupational lead, and to explore the effect of occupational lead exposure on BMD, so as to provide basis for the prevention and treatment of occupational lead poisoning and osteoporosis.Methods:An occupational health survey was conducted in a lead-acid battery enterprise in Jiangsu Province in January 2019. Basic information and occupational health examination results of 402 persons exposed to occupational lead were collected, and BMD was measured. Spearman rank correlation test was used to analyze the relationship between blood lead and BMD, and logistic regression analysis was used to analyze the influencing factors of BMD.Results:The blood lead level M ( P25, P75) of 402 occupational lead exposure workers was 220.5 (118.0, 307.0) μg/L, 46 workers (11.4%) had blood lead value ≥400 μg/L, and 5 workers (1.2%) ≥600 μg/L. 124 workers (30.8%) had abnormal BMD. The concentrations of lead dust and lead smoke in the workplace were <0.004-0.027 and <0.021-0.045 mg/m 3, respectively. The positions exceeding the standard point were mainly concentrated in the casting and welding group (44.4%, 4/9) of lead smoke positions. There was a statistically significant difference in the overall distribution of blood lead levels among lead exposure workers with different BMD levels, and there was a positive correlation between blood lead and BMD ( P<0.01) . The results of univariate analysis showed that there were statistically significant differences in the distribution of abnormal BMD among workers exposed to different genders, positions and blood lead levels ( P<0.01) . The results of multivariate logistic regression analysis showed that the risk of abnormal BMD in male workers was 5.069 times of that in female worker (95% CI: 2.906-8.840, P<0.01) . Conclusion:Occupational lead exposure personnel have a high blood lead level and a high abnormal BMD rate. Exposure to lead working environment is an influencing factor for the abnormal BMD of workers, so enterprise managers should pay attention to health protection, occupational health monitoring and supervision of working environment of front-line workers.

Objective:To investigate the effect of cocaine- and amphetamine-regulated transcript peptide (CART) on the synapse structure of mice cortical neuron subjected to oxygen-glucose deprivation (OGD).Methods:Primary neurons of the embryonic cerebral cortex obtained from healthy and clean Kunming mice at gestational age of 16-17 d were cultured. They were divided into control group, CART group, OGD group, and OGD+ CART group. 0.4 nmol/L CART 55-102 was added and cultured for 12 h after OGD treatment in the OGD+ CART group; the CART group was given the same dose of CART 55-102. The neuronal mortality was measured by the flow cytometry. The changes of synaptic structure were observed by immunofluorescence analysis, and the axon length and synapsin Ⅰ positive area were quantitatively analyzed. Real-time fluorescent quantitative polymerase chain reaction and Western blot analysis were used to identify the brain-derived neurotrophic factor (BDNF) mRNA and protein expression. Results:Compared with the control group, the mortality of neurons in the OGD group was significantly increased, the neuronal synapse growth was significantly inhibited, the positive area of synapsin Ⅰ was significantly reduced, and the expression levels of BDNF mRNA and protein were significantly down-regulated (all P<0.05). Compared with the OGD group, adding CART 55-102 significantly reduced the mortality of OGD neurons ( P<0.05), reversed the inhibitory effect of OGD on neuronal synapse growth, significantly increased the length of neuron axons and the positive area of synapsin Ⅰ (all P<0.05), and significantly up-regulated BDNF mRNA and protein expression levels (all P<0.05). Conclusion:CART can protect the synaptic structure of mice cortical neuron subjected to OGD, and its mechanism may be related to the up-regulation of BDNF expression.

T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
Chickens ; Clone Cells ; Cycloheximide ; Cytoplasmic Granules ; Humans ; Protein Biosynthesis ; RNA Precursors ; RNA-Binding Proteins ; T-Lymphocytes