Real-world pharmacological treatment of pregnant patients with rheumatic diseases from China: a retrospective analysis from 2016 to 2021.

Introduction: We investigated trends in the use of therapeutic drugs for pregnant patients with rheumatic diseases in nine Chinese cities (Beijing, Chengdu, Guangzhou, Harbin, Hangzhou, Shanghai, Shenyang, Tianjin, and Zhengzhou) to provide a reference for drug use in clinic. Methods: Outpatient prescription data for pregnant patients diagnosed with rheumatic diseases in nine cities across China in 2016-2021 were extracted from the Hospital Prescription Cooperation Project of the Hospital Pharmacy Professional Committee of the Chinese Pharmaceutical Association. A retrospective analysis was then performed, incorporating data on patient age, defined daily doses (DDDs), defined daily cost (DDC), and other metrics. Results: In 2016-2020, more than 70% of the pregnant patients diagnosed with rheumatic diseases in these nine cities were 25 to < 35 years of age. The most common rheumatic diseases during pregnancy were antiphospholipid antibody syndrome (APS) and systemic lupus erythematosus (SLE). In terms of the routine use of daily therapeutic drugs, the DDDs of low molecular weight heparins (LMWHs), glucocorticoids, and immunosuppressive agents dominated the top three. Intravenous immunoglobulin (IVIG) and tumor necrosis factor inhibitors (TNFi) have been used since 2019 and had been in the forefront of the DDC. Conclusion: The number and total cost of prescriptions for therapeutic drugs of pregnancy complicated by rheumatic diseases, have increased significantly over the study interval. Conventional therapeutic drugs, especially glucocorticoids, LMWHs, and hydroxychloroquine were the most widely used drugs in pregnant patients with rheumatic diseases. However, IVIG and TNFi, relatively high cost, have shown gradual increases in clinical use since 2019.

Toxic effects and transcriptional responses in zebrafish liver cells following perfluorooctanoic acid exposure.

As a typical type of persistent organic pollutant, perfluorooctanoic acid (PFOA) is pervasive in the environment. Multiple studies have found that PFOA has hepatotoxicity, but the mechanism remains poorly understood. In this study, the toxic effects of different concentrations of PFOA on zebrafish liver cells were systematically assessed by recording cell survival, ultrastructural observations, and transcriptome analyses. The results showed that the inhibition of cell viability and the massive accumulation of autophagic vacuoles were observed at 400 µM PFOA, while transcriptomic changes occurred with treatments of 1 and 400 µM PFOA. The transcription levels of 1055 (977 up- and 78 down-regulated genes) and 520 (446 up- and 74 down-regulated genes) genes were significantly changed after treatment with 1 and 400 µM PFOA, respectively. Based on Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, significant expression changes were observed in autophagy, tight junction, signal transduction, immune system, endocrine system, and metabolism-related pathways, indicating that such processes were greatly affected by PFOA exposure. The findings of this study will provide a scientific basis for the toxic effects and potential toxic mechanisms of PFOA on zebrafish, and provide information for ecological risk assessments.

Sodium hydrosulfide prevents myocardial dysfunction through modulation of extracellular matrix accumulation and vascular density.

The aim was to examine the role of exogenous hydrogen sulfide (H2S) on cardiac remodeling in post-myocardial infarction (MI) rats. MI was induced in rats by ligation of coronary artery. After treatment with sodium hydrosulfide (NaHS, an exogenous H2S donor, 56 µM/kg·day) for 42 days, the effects of NaHS on left ventricular morphometric features, echocardiographic parameters, heme oxygenase-1 (HO-1), matrix metalloproteinases-9 (MMP-9), type I and type III collagen, vascular endothelial growth factor (VEGF), CD34, and α-smooth muscle actin (α-SMA) in the border zone of infarct area were analyzed to elucidate the protective mechanisms of exogenous H2S on cardiac function and fibrosis. Forty-two days post MI, NaHS-treatment resulted in a decrease in myocardial fibrotic area in association with decreased levels of type I, type III collagen and MMP-9 and improved cardiac function. Meanwhile, NaHS administration significantly increased cystathionine γ-lyase (CSE), HO-1, α-SMA, and VEGF expression. This effect was accompanied by an increase in vascular density in the border zone of infarcted myocardium. Our results provided the strong evidences that exogenous H2S prevented cardiac remodeling, at least in part, through inhibition of extracellular matrix accumulation and increase in vascular density.

[Treatment of sludge liquor produced in deep dehydration by photoelectro-Fenton process].

Photoelectro-Fenton process was developed to treat the sludge liquor produced in deep dehydration. The results indicated that pollutants could be removed efficiently from the sludge liquor. Under the optimum condition of pH 3.0, H2O2 concentration of 65.3 mmol x L(-1), FeSO4 concentration of 6.53 mmol x L(-1) [ n(Fe2+): n (H2O2) = 1:10], 7.5V, the COD removal efficiency reached 59.0 % after 20 min treatment. The removal efficiency of TOC, TN, NH: -N and TP could reach 49. 3% , 20. 6% , 73.6% and 96.5% , respectively. This study would provide the reference for photoelectro-Fenton process in real wastewater treatment.

[Effects of the rich selenium-banqiao-Codonopsis pilosula on the aged rats' immune functions and its underlying mechanism].

OBJECTIVE: To study the effects of the Rich Selenium-Banqiao-Codonopsis Pilosula (BCPA) injecta on the aged rats' immune functions and its underlying mechanism. METHODS: Totally 60 rats, composed of 2, 12 and 22 month age old (half male and half female), were served as a young group, middle-age group and aged group respectively. Each group rats were randomly divided into the control and the BCPA subgroup (n = 10). The BCPA group was injected with BCPA at 7.2 g/kg intraperitoneally every day and the control group was injected the same volume of normal saline. All rats were conventionally fed for 45 days. An immune injection was performed after 15 days of BCPA injection. On the 22nd day, late-onset immune response would be induced. The caudal vein blood was collected and the antigen specific IgG, IgG1 and IgG2a antibody was detected on the 15th, 30th and 45th day. On the 45th day, the major T cell subgroups of splenic cells were analyzed and splenic cells were proliferated. RESULTS: No significant difference in the delayed-type hypersensivity (DTH) reaction was found between the control and the BCPA subgroups in the young and middle-aged rats while the aged BCPA subgroup had a stronger DTH reaction. There was no significant difference in the blood content of specific IgG, IgG1 and IgG2a antibody between the young and middle-age BCPA group while the aged BCPA group rats had an obvious enhancing reaction to the three antibodies mentioned above (P < 0.05). There was no obvious difference in the number of the CD3+ lymphocytes and the CD4+ T helper lymphocytes between the control and the BCPA subgroup in the young aged rats while a significant increase was spotted between the middle-aged and the aged group (P < 0.05). The splenic cells from young BCPA group rats had a strong proliferation response (P < 0.05). CONCLUSION: BCPA can enhance DTH reaction, potentiate the production of specific IgG, IgG1 and IgG2a antibody to resist KLH, improve the reaction to antigen, increase the amount of CD4+ cell, promote the immune response and had an important role in anti-immunosenescence and antioxidant capacity improvement in the aged rats.

Daphnoretin-induced apoptosis in HeLa cells: a possible mitochondria-dependent pathway.

Daphnoretin is a bicoumarin compound isolated from a natural product, Wikstroemia indica, which has been used to treat many diseases. It has strong antiviral and anti-tumor activities. Taking the anti-tumor activity of daphnoretin as a starting point, the present study aimed to test the pro-apoptotic effect of daphnoretin and its underlying mechanism in HeLa cells. The inhibitory effects of daphnoretin on viability and proliferation of HeLa cells were determined by the MTT assay. Daphnoretin-induced apoptotic morphological changes were analyzed by mitochondrial membrane potential and Hoechst staining. The number and stage of apoptotic HeLa cells were determined by flow cytometry. Gene expression was determined by reverse-transcription polymerase chain reaction. Protein expression was determined by western blot. The caspase activity of HeLa cells was detected by a caspase-3 and caspase-9 colorimetric assay kit. We found that daphnoretin significantly inhibited HeLa cells' viability by the MTT assay and flow cytometry. The nuclei of the apoptotic cells exhibited strong, blue fluorescence in Hoechst staining. Bax mRNA and protein levels were increased while bcl-2 mRNA levels were decreased after daphnoretin treatment. Daphnoretin also activated both caspase-3 and caspase-9. These findings suggest that daphnoretin promotes apoptosis of HeLa cells in a mitochondria-mediated way. Daphnoretin therefore has potential to be a promising drug to treat uterine cervix cancer.

Molecular cloning, subcelluar location and expression profile of signal transducer and activator of transcription 2 (STAT2) from turbot, Scophthalmus maximus.

Signal transducer and activator of transcription 2 (STAT2) is an important molecule involved in the type I interferon signalling pathway. To date, little STAT2 homologue is available in fish except Atlantic salmon and goldfish. In this paper, STAT2 was firstly cloned and characterized from turbot, a marine flatfish with high economic value. Briefly, turbot STAT2 cDNA is 3206 bp in length encoding a predicted protein of 793 amino acids. The phylogenetic tree shows that turbot STAT2 protein shared the closest relationship with Atlantic salmon. Analysis of subcellular distribution indicates that STAT2 is mainly present in the cytoplasm of TK cells. Stat2 mRNA is constitutively expressed in widespread tissues and induced by several folds in turbot tissues and TK cells after stimulation with Vibrio anguillarum and lymphocystis disease virus (LCDV). Unlike the higher vertebrate STAT2, turbot STAT2 nuclear export signal (NES) exists not in the C-terminal 79 amino acids but in N-terminal 137-312 amino acids (STAT_alpha domain). The nuclear translocation of turbot STAT2 after Poly(I:C) treatment proved its transcription activity in TK cells. All these results suggested that STAT2 may be involved in the immune response in turbot as a transcription factor.

Evaluation of reference genes for quantitative real-time RT-PCR analysis of gene expression in Nile tilapia (Oreochromis niloticus).

Quantitative real-time reverse-transcriptase polymerase chain reaction (RT-qPCR) has been used frequently to study gene expression related to fish immunology. In such studies, a stable reference gene should be selected to correct the expression of the target gene. In this study, seven candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase (GADPH), ubiquitin-conjugating enzyme (UBCE), 18S ribosomal RNA (18S rRNA), beta-2-microglobulin (B2M), elongation factor 1 alpha (EF1A), tubulin alpha chain-like (TUBA) and beta actin (ACTB)), were selected to analyze their stability and normalization in seven tissues (liver, spleen, kidney, brain, heart, muscle and intestine) of Nile tilapia (Oreochromis niloticus) challenged with Streptococcus agalactiae or Streptococcus iniae, respectively. The results showed that all the candidate reference genes exhibited tissue-dependent transcriptional variations. With PBS injection as a control, UBCE was the most stable and suitable single reference gene in the intestine, liver, brain, kidney, and spleen after S. iniae infection, and in the liver, kidney, and spleen after S. agalactiae infection. EF1A was the most suitable in heart and muscle after S. iniae or S. agalactiae infection. GADPH was the most suitable gene in intestine and brain after S. agalactiae infection. In normal conditions, UBCE and 18S rRNA were the most stably expressed genes across the various tissues. These results showed that for RT-qPCR analysis of tilapia, selecting two or more reference genes may be more suitable for cross-tissue analysis of gene expression.

Screening and analysis of PoAkirin1 and two related genes in response to immunological stimulants in the Japanese flounder (Paralichthys olivaceus).

A member of the NF-κB signaling pathway, PoAkirin1, was cloned from a full-length cDNA library of Japanese flounder (Paralichthys olivaceus). The full-length cDNA comprises a 5'UTR of 202 bp, an open reading frame of 564 bp encoding a 187-amino-acid polypeptide and a 521-bp 3'UTR with a poly (A) tail. The putative protein has a predicted molecular mass of 21 kDa and an isoelectric point (pI) of 9.22. Amino acid sequence alignments showed that PoAkirin1 was 99% identical to the Scophthalmus maximus Akirin protein (ADK27484). Yeast two-hybrid assays identified two proteins that interact with PoAkirin1: PoHEPN and PoC1q. The cDNA sequences of PoHEPN and PoC1q are 672 bp and 528 bp, respectively. Real-time quantitative reverse-transcriptase polymerase chain reaction analysis showed that bacteria could induce the expressions of PoAkirin1, PoHEPN and PoC1q. However, the responses of PoHEPN and PoC1q to the bacterial challenge were slower than that of PoAkirin1. To further study the function of PoAkirin1, recombinant PoAkirin1 and PoHEPN were expressed in Escherichia coli and would be used to verify the PoAkirin1-PoHEPN binding activity. These results identified two proteins that potentially interact with PoAkirin1 and that bacteria could induce their expression.

Iron-metabolic function and potential antibacterial role of Hepcidin and its correlated genes (Ferroportin 1 and Transferrin Receptor) in turbot (Scophthalmus maximus).

Antimicrobial peptide plays an important role in fish immunity. The small molecular antimicrobial peptide Hepcidin in turbot was studied and reported in this paper. The Ferroportin 1 (FPN1) and Transferrin Receptor (TFR) genes, which are related to Hepcidin, were cloned in turbot. The characteristics of Hepcidin and its related genes were studied, including an analysis of the expression patterns and cloning of the Hepcidin promoter, the relationship between Hepcidin and NF-κB and the regulation of iron-metabolism. The results showed that the promoter of SmHepcidin contains the binding sites of NF-κB, and NF-κB may directly or indirectly receive feedback signals from SmHepcidin. In the liver, spleen and kidney, in which there was an increased SmHepcidin expression level, SmFPN1 dramatically decreased and SmTFR was also either decreased or exhibited no obvious change after bacterial/viral infection and an injection of exogenous Hepcidin protein. RNAi experiments in turbot kidney cells confirmed the expression changes of these gene patterns. Furthermore, the administration of exogenous Hepcidin protein, which regulates the level of chelatable iron in cells, further confirmed the function of Hepcidin in iron metabolism. It is speculated that the rapidly increased expression of SmHepcidin may induce changes in the expression of related genes, and that the in vivo chelatable iron concentration which participates in the antibacterial process was also changed when exogenous pathogens are present in turbot. It is suggested that SmHepcidin plays a defensive role against pathogenic infection.

[Corrosion resistance of casted titanium by compound treatments in the artificial saliva with different fluoride concentrations].

OBJECTIVE: To study the corrosion resistance of casted titanium by plasma nitriding and TiN-coated compound treatments in the artificial saliva with different fluoride concentrations and to investigate whether compound treatments can increase the corrosion resistance of casted titanium. METHODS: Potentiodynamic polarization technique was used to depict polarization curve and to measured the current density of corrosion (Icorr) and the electric potential of corrosion (Ecorr) of casted titanium (Group A) and casted titanium by compound treatments (Group B) in the artificial saliva with different fluoride concentrations. After electrochemical experiment, the microstructure was observed by scanning electron microscope (SEM). RESULTS: The Icorrs of Group A and B in the artificial saliva of different fluoride concentrations were (1530.23 ± 340.12), (2290.36 ± 320.10), (4130.52 ± 230.17) nA and (2.62 ± 0.64), (7.37 ± 3.59), (10.76 ± 6.05) nA, respectively. The Ecorrs were (-0.93 ± 0.10), (-0.89 ± 0.21), (-0.57 ± 0.09) V and (-0.21 ± 0.04), (-0.17 ± 0.03), (-0.22 ± 0.03) V, respectively.The Icorrs of Group B were significantly lower (P < 0.01)than that of Group A. The Icorrs increased significantly with the increasing of fluoride concentrations (P < 0.01). The Ecorrs of Group B were significantly higher than that of Group A (P < 0.01). The SEM confirmed the microstructure in the casted titanium was much severely than that in Group B, the microstructure in Group A and B corroded more and more heavily with increasing of fluoride concentrations. CONCLUSIONS: The increase of fluoride concentrations influence the corrosion resistance of both treated and untreated casted titanium negatively, but plasma nitriding and TiN-coated compound treatments can significantly increase the corrosion resistance of casted titanium.

Cloning, characterization and expression analysis of a cold shock domain family member YB-1 in turbot Scophthalmus maximus.

The Y-box proteins are a family of highly conserved nucleic acid binding proteins. In this report we have identified a new member, YB-1 from turbot (Scophthalmus maximus) spleen cDNA library. The full-length cDNA sequence of turbot YB-1 was obtained and then the expression at transcriptional level was researched by qRT-PCR. In normal organs, the expression of YB-1 was higher in liver, brain, gill and heart, respectively. YB-1 had the highest expression level at gastrula stage during the early stages of embryo development. In the liver, kidney and spleen, the turbot YB-1 expression level was the highest at 72 h after challenge with lymphocystis disease virus (LCDV) and the highest at 12 h after challenge with Vibrio anguillarum (V. anguillarum). Furthermore, the expression of turbot YB-1 also distinctly increased in turbot kidney cells (TK) at 24 h after challenge with V. anguillarum and LCDV. These results indicated that the turbot YB-1 protein may play a significant role in the immune response of turbot.

A new Akirin1 gene in turbot (Scophthalmus maximus): molecular cloning, characterization and expression analysis in response to bacterial and viral immunological challenge.

SmAkirin1, a member of the NF-κB signaling pathway, was isolated from turbot by RACE. Its cDNA was 564 bp and encoded a putative protein of 187 amino acids with a predicted molecular mass of 21 kDa and an isoelectric point (pI) of 9.05. Amino acid sequence alignments showed that SmAkirin1 was 91% identical to the Salvelinus alpinus Akirin1 protein ACV49694. Transient expression of SmAkirin1-GFP in the turbot kidney cell line SMKC revealed a nuclear localization of the protein, and a typical NLS signal was found at the N-terminal region of the SmAkirin1 protein. Trans-activation assay in yeast demonstrated that SmAkirin1 has no transcriptional activation. Transcriptional analysis showed that SmAkirin1 was expressed in all of the tissues examined, with the highest expression in the spleen and brain. Real-time quantitative reverse-transcriptase polymerase chain reaction analysis showed that the SmAkirin1 transcript was induced by bacterial and viral infection.

Signal transducer and activator of transcription 3 (STAT3) homologue in turbot (Scophthalmus maximus): molecular characterization and expression analysis.

Signal transducer and activator of transcription 3 (STAT3) acts as an important mediator in multiple biological processes induced by different cytokines. So far, little information is available in fish STAT3. In this study, turbot (Scophthalmus maximus) STAT3 gene was cloned and characterized for the first time. The turbot STAT3 full-length cDNA consists of 2355 nucleotides encoding a polypeptide of 784 amino acids with four conserved domains including STAT_int, STAT_alpha, STAT_bind and SH2 domain. The phylogenetic tree showed that turbot STAT3 shared the closest relationship with mandarin fish (Siniperca chuatsi) STAT3. The autoactivation experiment in yeast proved that turbot STAT3 was a strong transcription factor. The quantitative RT-PCR experiment indicated that Stat3 mRNA was expressed in widespread tissues with the highest expression levels in the liver. And the further expression patterns analysis revealed that turbot Stat3 expression levels were increased in liver, spleen, kidney of fish infected with Vibrio anguillarum and liver of fish infected with LCDV. Meantime, hepcidin, one of STAT3 target gene, was also up-regulated in liver of fish infected with two pathogens. These results suggested that turbot Stat3 may involved in the immune defense process as a transcription factor.

Comparison of chromosome preparation methods for the different developmental stages of the half-smooth tongue sole, Cynoglossus semilaevis.

Here we present a comparison of the different chromosome preparation methods, including the method of the single embryo or larva, the method of juvenile-swimming and the method of phytohemagglutinin injection, used for the different developmental stages of the half-smooth tongue sole, Cynoglossus semilaevis. The mean index of mitosis for the three methods was 0.79%, 0.09% and 0.15%, respectively. From the well-spread metaphase chromosomes obtained, it was determined that C. semilaevis has a diploid chromosome number of 42 and heterotypic sex chromosomes. Also, the metaphase chromosomes produced could be used for karyotype preparation and banding studies. The three methods are also successfully used for chromosome preparation in Japanese flounder (Paralichthys olivaceus) and summer flounder (Paralichthys dentatus).

Protective effects of cysteine analogues on acute myocardial ischemia: novel modulators of endogenous H(2)S production.

The current study was designed to evaluate the pharmacologic effects of three novel cysteine-containing compounds: S-propyl-l-cysteine (SPC), S-allyl-l-cysteine (SAC), and S-propargyl-l-cysteine (SPRC) on H(2)S production and antioxidant defenses in an acute myocardial infarction (MI) rat model. The enzymatic activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), as well as glutathione redox status and malonaldehyde (MDA) content, also were determined. All three compounds were found to preserve SOD and GPx activities and also tissue GSH levels while reducing the formation of the lipid peroxidation product MDA in ventricular tissues. With immunfluorescence assays, we observed the expression of CSE and Mn-SOD. The morphologic changes of the cardiac cells are seen with both light and electron microscopy. The corresponding pathologic alterations were characterized mainly as loss of adherence between cardiac myocytes and swollen or ruptured mitochondria at the ultrastructural level. Propargylglycine, a selective inhibitor of CSE, abolished the protective effects of each compound used in the current model. Our study provides novel evidence that SPC, SAC, and SPRC have cardioprotective effects in MI by reducing the deleterious effects of oxidative stress by modulating the endogenous levels of H(2)S and preserving the activities of antioxidant defensive enzymes like SOD.

Hydrological adjustment and flooding control of wetlands in the Liaohe Delta.

The function of estuary wetland on hydrological adjustment and flooding control is studied in this paper. It is estimated that the evapotranspiration in the reed field during growth season (June to October) is 722.9 mm, which is 37.5% higher than large water body (E601: 525.9 mm). The water replacement rate in the reed field can reach 95% only when the rains continuously for 11 days and the precipitation reached 912 mm. For the water balance in the paddy field, the total water requirement ranges between 1920 and 1860 mm, among which, 31% is from precipitation, and the left is provided by reservoirs. The water usage efficiency is 0.35 at present productivity. Based on the landscape characteristics and functionalities on flooding control, 5 functional zones are designed for the Liaohe Delta; key protected area; underground storage area; flooding discharge area; flood diversion area in emergency; and flood control drainage area.