Mechanism of action of formononetin in alleviating allergic asthma through DRP1-NLRP3 signaling pathway / 中国药理学通报

Aim To investigate the mechanism by which formononetin (FN) inhibits mitochondrial dynamic-related protein 1 (DRP1) -NLRP3 axis via intervening the generation of ROS to reduce allergic airway inflammation. Methods In order to establish allergic asthma mouse model, 50 BALB/c mice aged 8 weeks were divided into the control group, model group, FN treatment group and dexamethasone group after ovalbumin (OVA) induction. Airway inflammation and collagen deposition were detected by HampE and Masson staining. Th2 cytokines and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and IgE levels in bronchoalveolar lavage fluid (BALF) were measured by ELISA, ROS in BEAS-2B cells was assessed by DCFH-DA staining, DRP1 expression in lung tissue and BEAS-2B cells was detected by immunohistochemistry and immunofluorescence, and the DRP1-NLRP3 pathway was analyzed by immunoblotting. Results FN treatment could effectively ameliorate the symptoms of asthmatic mouse model, including reducing eosinophil accumulation, airway collagen deposition, decreasing Th2 cytokine and IgE levels, reducing ROS and MDA production, increasing SOD and CAT activities, and regulating DRP1-NLRP3 pathway-related protein expression, thereby relieving inflammation. Conclusion FN ameliorates airway inflammation in asthma by regulating DRP1-NLRP3 pathway.

Summary of best evidence for the management of thirst in ICU patients / 中国实用护理杂志

Objective:To summarize the best evidence of thirst management in ICU patients and provide evidence-based basis for dinical practice.Method:According to the "6S" evidence pyramid model, the literature on thirst management of ICU patients was systematically retrieved from relevant guidelines websites, evidence-based databases, association websites and original literature databases at home and abroad. The retrieval time was from the establishment of the database to June 31, 2022. Two researchers with evidence-based nursing training independently completed literature quality evaluation. To extract and summarize the evidence of the literature that meets the quality standard.Results:A total of 17 articles were included, including 8 randomized controlled trials, 5 quasi-experimental studies and 4 cross-sectional studies. The 18 pieces of best evidence were formed, including 5 aspects: basic requirements of thirst management, intervention evaluation, intervention methods, matters needing attention and health education.Conclusions:This study summarized the best evidence of thirst management in ICU patients. Nurses should translate and apply the best evidence in combination with the clinical situation and specific policies of the department to relieve the thirst symptoms of ICU patients.

Panax notoginseng saponin Rl attenuates allergic rhinitis through AMPK/DRP1 mediated mitochondrial fission / 中国药理学通报

Aim To investigate whether notoginsenoside Rl (PNS-R1) alleviates allergic rhinitis (AR) through AMP-activated protein kinase (AMPK)/mitochondrial fission critical protein (DRP1) -mediated mitochondrial fission. Methods Different doses of PNSRl were used to treat ovalbumin (OVA) -induced AR model mice,and the inhibitory effect of PNS-R1 on AR was investigated by observing allergic symptoms such as nasal rubbing and sneezing, as well as HE staining of nasal tissues. Serum IgE levels and nasal lavage fluid (NLF) inflammatory cytokine levels were detected by enzyme-linked immunosorbent assay (ELISA) and apoptosis-related proteins were detected by Western blot. In vitro human nasal epithelial cells (HNEpC) were stimulated with IL-13 to observe apoptosis, mitochondrial membrane potential, cellular ROS and mitochondrial ROS production, as well as the expression levels of AMPK/DRP1, expression levels of the TXNIP/NLRP3 inflammasomes and the translocation of DRP1. Results PNS-R1 attenuated allergic symptoms in AR mice, HE staining reduced inflammatory cells and reduced the levels of OVA-specific IgE in serum, and the levels of IL-4, IL-6, and IL-8 in NLF. PNS-R1 attenuated the apoptosis and ROS production of nasal epithelial cells in AR. In vitro PNS-R1 could up-regulate mitochondrial membrane potential after IL-13 stimulation, reduce ROS and mtROS production, the proportion of apoptotic positive cells, and reduce cleaved caspase-3, Bax, and up-regulate Bcl-2 expression, down-regulate DRP1 phosphorylation (Ser 616) and DRP1 translocation at the mitochondrial membrane in an AMPK-dependent manner, reducing TXNIP/NLRP3 expression. Conclusions PNS-R1 can protect mitochondrial integrity by inhibiting the AMPK/DRP1 signaling axis and its subsequent TXNIP/NLRP3 signaling axis,thereby alleviating rhinitis in AR mice.

JTE-013 mediates RhoA/ROCKl/Drpl signaling axis regulating mitochondrial damage and apoptosis to alleviate allergic rhinitis / 中国药理学通报

Aim To investigate the protective effect and mechanism of JTE-013 on allergic rhinitis (AR) by regulating mitochondrial injury and apoptosis through RhoA/ROCKl/Drpl pathway. Methods AR model was established by ovalbumin (OVA) in mice. Nasal tissue sections were then stained with HE, TUNEL and DHE. Western blot assay. In vitro, human nasal epithelial cells (HNEpCs) were stimulated with human recombinant interleukin-13 (IL-13), and the effects of JTE-013 and Y27632-related protein expression were detected by Western blot. Immunofluorescence was used to observe the effects of JTE-013 and Y 27632 on total ROS, mitochondrial membrane potential and mitochondrial ROS generation, Drpl translocation and Cyt-c expression in cells. Results JTE-013 reduced the frequency of nose rubbing and sneezing, reduced nasal mucosal thickening and decreased eosinophil infiltration in AR mice. TUNEL and DHE staining results suggested that JTE-013 could inhibit apoptosis and reduce ROS expression in mouse nasal epithelial cells. Western blot showed that both JTE-013 and Y 27632 could significantly reduce RhoA, ROCK1, Drpl and p-Drpl(616), inhibit the expression of apoptotic proteins Bax, cleaved-caspase-3, Cyt-c, cleavedcaspase-9 and up-regulate the expression of p-Drpl (637) and Bcl-2. Immunofluorescence showed that inhibitors of JTE-013 or ROCK1 almost blocked IL-13mediated increase in ROS and mtROS production, inhibited decrease in mitochondrial membrane potential, and blocked Cyt-c expression and Drpl translocation in nasal mucosal epithelial cells. Conclusion JTE-013 can regulate the morphology and function of mitochondria by inhibiting RhoA/ROCKl/Drpl signaling axis, thereby alleviating nasal epithelial cell inflammation in mice with allergic rhinitis.

Polydatin alleviates allergic rhinitis by regulating PINK1-Parkin-mediated mitophagy / 中国药理学通报

Objective To investigate the protective effect and mechanism of polydatin ( PD) on allergic rhinitis (AH) by regulating mitophagy through PINK1- Parkin signaling pathway, and to provide a new target for clinical treatment of AH.Methods Thirty-two BALB/c murine were randomly divided into 4 groups: control group, OVA group, PD low-dose (30 mg • kg 1 ) and high-dose ( 45 mg • kg 1 ) treatment groups.At the end of modeling, the total number of sneezing and nasal nibbing of murine was recorded.HE staining was used to observe the morphology of na- sal mucosal epithelium and eosinophil infiltration.Western blot was used to detect the expression of P1NK1 , Parkin, TOM20 and mitochondrial apoptosis- related proteins Bax, Bcl-2, caspase-3, cleaved- caspase-3 and Cytochrome C.Hie expression of P1NK1 and cleaved-caspase-3 in nasal epithelial cells (HNEpC) was observed by immunofluorescence.Re¬sults The frequency of sneezing and nasal rubbing movements was significantly increased, the nasal mu¬cosa epithelium was thickened, and eosinophils were accumulated in AH murine , these results were reversed after PD treatment.Western blot results shower] that signaling proteins PINK1/Parkin anrl pro-apoptotie pro¬teins, including Bax, caspase-3, cleaved-caspase-3 and Cytochrome C were significantly overexpressed, the expression of TOM20 and Bcl-2 was decreased in OVA group, and PD up-regulated the levels of P1NK1 , Parkin, as well as Bcl-2 and inhibited the expression of TDM20 and pro-apoptotic proteins, while after pre- treatment with mitochondrial division inhibitor 1 ( Mdi- vi-1 ) , the expression of P1NK1 and Parkin was re¬duced , the expression of TOM20 was increased, while PD treatment did not significantly affect this effect.From the immunofluorescence results, it can be seen that the level of P1NK1 was increased after IL-13 stim¬ulation of HNEpCs compared with the control group, and PD further up-regulated the expression of P1NK1 , which was suppressed after pretreatment with Mdivi-1 , while PD did not change this phenomenon.Western blot results for P1NK1 and cleaved-caspase-3 were con¬firmed by immunofluorescence.Conclusion PD may activate mitophagy through the P1NK1 -Parkin signaling pathway, thereby protecting against AR.

Identification and analysis of the R2R3-MYB transcription factor based on a transcriptome database in Morinda officinalis / 药学学报

In order to explore MYB transcription factors related to developmental processes and secondary metabolism in Morinda officinalis, we analyzed MoMYB expression based on transcriptome data from three tissues (root, stem and leaf). We used this analysis to provide a theoretical foundation for regulating the metabolism of M. officinalis. RNA-seq data along with the five databases including PFAM and plantTFDB and others were used to screen and classify MoMYB, including GO functional annotation and classification, subcellular localization, signal peptide prediction, conserved motif discovery, and comparative phylogenetic analysis. RT-qPCR was carried out to detect tissue-specific expression differences of MoMYB genes. According to transcriptome data, 109 MoMYB sequences were identified and divided into four classes, containing 51 sequences related to R2R3-MYB. Subcellular localization analysis indicated that a majority of sequences were located in nucleus. Blast2GO analysis showed that 109 MoMYB sequences were classified into three major functional ontologies including molecular function (112), biological processes (76) and cellular components (239). The R2-MYB conserved motif of 51 R2R3-MYB sequences possessed three significantly conserved tryptophan residues, whereas a phenylalanine replaced the first tryptophan in R3-MYB. The results of multiple sequence alignment and phylogenetic analysis revealed that the R2R3-MYB was distributed in all subgroups, apart from the S10, S19 and S21 subgroups. RT-qPCR indicated that several R2R3-MYB genes were differentially expressed among the three tissues, and this finding was consistent with transcriptome data. The 109 MoMYB sequences were annotated and divided into different classes, which lays the foundation for further study on MYB transcriptional factors in M. officinalis.

Cloning and analysis of the DXR gene and its promoter in Morinda officinalis / 药学学报

The objective of this research was to clone 1-deoxy-D-xylulose 5-phosphate reductoisomerase gene (MoDXR) and its promoter sequence from Morinda officinalis and carry out bioinformatic analysis, cis-acting elements analysis, and prokaryotic expression. On the basis of the MoDXR gene sequence obtained from the M. officinalis transcriptome and with NCBI-ORFfinder analysis, a pair of specific primers were designed, and used for RT-PCR amplification. The promoter region sequence at the 5′ end of MoDXR gene was isolated by the genome walking technique. Localization of MoDXR was carried out by subcellular analysis. The prokaryotic expression plasmid pET-28a-MoDXR was constructed and transfected into Escherichia coli BL21(DE3) chemically-competent cells; the recombiant plasmid expressed fusion protein after the induction by IPTG. The full-length cDNA of MoDXR was 2 015 bp,and open reading frame (ORF) size was 1 425 bp, and it encoded 474 amino acid residues and had a molecular mass of 51.27 kD. Sequence comparison with BlastP to the NCBI database revealed that MoDXR had high sequence similarity with many other DXRs, such as Coffea arabica DXR (CaDXR) and Rauvolfia verticillata DXR (RvDXR). A phylogenetic tree revealed that MoDXR had its closest relationship with DXR from Coffea arabica and Gardenia jasminoides. The subcellular localization revealed that MoDXR protein was located on the chloroplast. Plantcare analysis indicated that the promoter region sequence of MoDXR was 1 493 bp, covering multiple light, stress, and hormone-responsive cis-regulatory elements; protein electrophoresis showed that the expressed protein was the anticipated size. This research lays the foundation for further purification and structural and functional characterization of the MoDXR protein.

Nasality Changes With Age in Normal Korean-Speaking Adults

OBJECTIVES: This study was performed to investigate the effects of aging on nasality and the influence of age-related changes in nasal cavity volume and nasal patency on nasality. METHODS: A total of 180 healthy Korean-speaking adult volunteers, who had no nasal or voice-related complaints, were enrolled in this study. Nasometry, acoustic rhinometry, and rhinomanometry were performed to obtain the nasalance score, nasal cavity volume, and nasal resistance, respectively. Changes in these parameters with age were analyzed. RESULTS: Nasal cavity volume increased significantly, and nasal resistance decreased significantly, with age. The nasalance scores for the nasal passage and oronasal passage decreased significantly with age, while there were no age-related changes in nasalance scores for the oral passage. CONCLUSION: Nasalance scores for the passages containing nasal consonants decreased with age although significant increases were observed in nasal cavity volume and nasal patency with age. Therefore, the age-related decreases in nasalance scores may result from factors other than changes in the nasal cavity.

Effect and mechanism of human dental pulp stem cells-derived exosomes on lipopolysaccharide induced acute lung injury / 军事医学

Objective To investigate the protective role and underlying mechanism of human dental pulp stem cells (DPSCs)-derived exosomes against lipopolysaccharide ( LPS) induced acute lung injury ( ALI) in pulmonary alveolar macrophage(PAM) cells of rats.Methods DPSCs were cultured in the complete culture medium , and their supernatants at passage 6 were collected after serum-free medium treatment for 24 hours.Exosomes were extracted and purified with ultracentrifugation .Rat PAM NR8383 was cultured in 12-well plate and treated with LPS of 1μg/ml alone or together with exosomes.The supernatants were then collected at 0, 6, 12 and 24 h respectively after treatment .Inflammatory cytokine levels of tumor necrosis factor-α(TNF-α)and interleukins (IL-1βand IL-6) in the supernatant were measured by ELISA assay and the expression and phosphorylation level of MAPK (p44/42), NF-κB and IκBαin cell lysates were detected with Western-blotting.Results Compared with control group , the content of TNF-α,IL-1βand IL-6 increased significantly in LPS group (P<0.05), which indicated that the inflammatory cell model was induced successfully .The levels of TNF-αand IL-1βwere obviously attenuated after a high doses of exosomes treatment (P<0.05), and the expression of IL-6 was markedly suppressed after low and high doses of exosomes treatment (P<0.05), compared with the group of LPS treatment alone.The phosphorylation of NF-κB, IκBαand p44/42 was significantly inhibited after treatment with the DPSCs-derived exosomes.Conclusion DPSCs-derived exosomes may have a potential protective effect on LPS-induced ALI, and the underlying mechanism is that the activity of MAPK (p44/42) and NF-κB/IκBαpathways are eliminated by DPSCs-derived exosomes.

Construction of full-length cDNA library of Sarcandra glabra and sequences analysis on EST / 中草药

Objective: Sarcandra glabra was recognized as an important research material attributing to its high medicinal value and economic value. However, little information was known about its genomics and regulatory pathway participating in reproductive development. For the first step to understand the molecular basis and further explore genes which related to metabolism and resistance in S. glabra. Methods: A SMART full-length complementary DNA library from the leaves tissue was constructed and characterized to providing the experimental basis for discovery of functional genes of S. glabra. The assembly expressed sequence tag (EST) data were completed by ABI3730 DNA program. A high quality full-length cDNA library was constructed successfully from S. glabra leaves. Results: The titer of library was 1.14×107 pfu/mL and the average length of inserted fragments was 1 000 bp. A total of 221 clones were sequenced from the cDNA library and obtained 177 EST sequences. The EST sequences were assembled into 151 unigenes including 12 contigs and 119 singletons (79%). EST exhibited significant similarity with known putative functional nucleotide sequences in the GenBank database. These genes were mostly involved in cell development, signal transduction, protein synthesis, transcription, stress tolerance response, energy metabolism based on molecular function of GO annotation. Conclusion: This report constructs a full-length-cDNA library and analyzes the bioinformatics of the related EST sequences, and then offers a reference to genomic research of S. glabra.

HIS-based analysis of clinical characteristics and combined treatment of Chinese and western medicine in diabetes deaths / 中国中药杂志

To understand the clinical characteristics and distribution of combined treatment of Chinese and western medicine in diabetes deaths, the hospitalization information of diabetes deaths from HIS system of 20 national 3A-grade general hospitals. Then the frequency statistics and association rules analysis were used to analyze the general information, complications, combined treatment, death time and other information of the patients died from diabetes. The results showed that most of the diabetes deaths were of middle aged and elderly people, more often in males than females. The complications with higher incidence included hypertension, pulmonary infection, coronary heart disease, cerebral infarction and renal inadequacy. In combined treatment rules, western medicines insulin, cefuroxime, furosemide, dopamine, nikethamide and sodium bicarbonate were used in combination at highest frequencies, followed by the combinations of traditional Chinese medicines panax notoginseng, radix bupleuri and western medicines, and the combinations between Chinese medicines had the lowest use frequency. Most of the diabetes deaths were of middle aged and elderly people, more often in males than females. They mainly died from 3 pm to 5 pm and from 5 pm to 7 pm. Therefore, the diabetes deaths often had complications of cardiovascular and cerebrovascular diseases, and early prevention shall be noted in clinics; the clinical treatment plan was basically in accordance with the guidelines for clinical treatment of diabetes; the drugs with promoting blood circulation to remove blood stasis and soothing liver-qi stagnation effects were the common Chinese medicines in treatment of diabetes.

Two Different Total Hip Arthroplasties for Hartofilakidis Type C1 Developmental Dysplasia of Hip in Adults / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Total hip arthroplasty (THA) in developmental dysplasia of the hip (DDH) is more complex than the normal hip, with large replacement risks and many complications. Although nonosteotomy THA is convenient to perform, femoral osteotomy shortening can avoid blood vessel and nerve traction injuries. This study aimed to compare osteotomy THA with nonosteotomy to determine reasonable options for operative management of DDH.</p><p><b>METHODS</b>Data on 48 DDH patients who underwent THA were analyzed retrospectively. The patients were divided into two groups: Group A 29 cases (nonosteotomy), and group B 19 cases (osteotomy). Harris and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores, limb length discrepancy (LLD), radiological data on the hip, and claudication were evaluated. Data were analyzed by using paired-sample Student's t-test, independent-sample Student's t-test, and Pearson's Chi-square test; the test level was α =0.05.</p><p><b>RESULTS</b>Postoperative Harris (90.7 ± 5.1) and WOMAC scores (88.0 ± 10.6) were significantly improved compared with preoperative Harris (44.8 ± 5.7) and WOMAC scores (42.0 ± 5.3) in group A (P < 0.05). Postoperative Harris (90.4 ± 2.8) and WOMAC scores (88.2 ± 5.9) were significantly improved compared with preoperative Harris (44.4 ± 4.2) and WOMAC scores (43.2 ± 4.3) in group B (P < 0.05). One case of dislocation occurred in group A; after closed reduction, dislocation did not recur. In group A, 2 patients developed cutaneous branch injury of the femoral nerve, which spontaneously recovered without treatment. Postoperative LLD >2 cm was seen in one case in group A and five cases in group B. Postoperative claudication showed no significant difference between the two groups (P > 0.05). No patients developed infection; postoperative X-rays showed that the location of the prosthesis was satisfactory, and the surrounding bone was not dissolved.</p><p><b>CONCLUSIONS</b>THA is effective and safe for DDH. For unilateral high dislocation DDH patients with limb lengthening ≤4 cm and good tissue conditions, THA without femoral osteotomy may be considered.</p>

Lower-limb valgus deformity associated with developmental hip dysplasia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Treating developmental dysplasia of the hip is often challenging. The difficulties include not only the hip surgery itself but also the treatment of the associated lower-limb valgus deformity. However, there have been very few studies on such deformity in patients with developmental hip dysplasia. In this study, we investigated the prevalence and severity of lower-limb valgus deformity, along with the relationship between the severity of valgus deformity and mechanical alterations of the hip or the ipsilateral knee.</p><p><b>METHODS</b>Two hundred and six affected lower limbs of 116 adult patients with untreated developmental dysplasia of the hip were included in the study, grouped according to the severity of hip dysplasia. Each study participant's radiographs were measured to quantitatively evaluate the mechanical axis deviation of the lower limb, and further to evaluate the prevalence and severity of the lower-limb valgus deformity. Some mechanical alterations of the hip and the ipsilateral knee were also measured on the radiographs.</p><p><b>RESULTS</b>Of the affected lower limbs, 14.1% had valgus deformities. Study participants with Crowe type III hip dysplasia had the most severe deformity and the highest prevalence of deformity. Severity of valgus deformity had a strong positive correlation with the lateral migration of the femoral head but not with the superior migration. A decreased lateral distal femoral angle contributed to the lower-limb valgus deformity, and the lateral distal femoral angle had a strong negative correlation with the severity of valgus deformity.</p><p><b>CONCLUSIONS</b>Hip dysplasia is commonly associated with lower-limb valgus deformity, and the severity of the lower-limb valgus deformity is mostly affected by lateral migration but not superior migration of the femoral head. The valgus deformity may originate mainly in the distal femur, in addition to the hip joint itself. These findings can be taken into account when planning to treat the patients with hip dysplasia.</p>

Study of cognitive function in patients with lacunar infarction associated with leukoaraiosis / 中国脑血管病杂志

Objective: To investigate the effect of different degrees of ischemic white matter lesions on the cognitive function in patients with lacunar infarction. Method: One hundred twelve consecutive patients with lacunar infarction were collected. Age-related white matter changes rating scale (ARWMCRs) was used, and according to the results of ARWMCRs scores, the patients were divided into mild (n = 34, 0-3 scores), moderate (n = 43, 4-7 scores), and severe (n = 35, 8-24 scores) groups. The national health institutes of stroke scale (NIHSS) and the modified Rankin scale (mRS) scores were used to assess the degree of neurological deficit. The mini-mental status scale (MMSE) and the Montreal Cognitive Assessment (MoCA) were used for neuropsychological tests, and event-related potential two-tone sequence auditory P300 (2t-P300) wave was used for neural electrophysiological examination. The cognitive function was evaluated. The cognitive function among all groups was compared. Results: Circled digit oneThere were significant differences in comparison of the MoCA scores among the three groups. Only the mRS score and the MMSE score in the severe group were lower than those in the mild and moderate groups. There were significant differences (P < 0.05). Circled digit twoThe latency of P300 wave in the severe group was higher than that in the moderate and mild groups, and the latency of P300 wave in the moderate group was higher than that in the mild group. There was significant difference (P <0.05). The amplitude of P300 wave in the severe group was lower than that in the moderate group and mild group. There were significant differences (P < 0.05). Circled digit threeThe ARWMCRs score in patients with white matter lesions was negatively correlated with the MMSE score, MoCA score, and amplitude of P300 wave. It was positively correlated with the latency of P300 wave and mRS score. Conclusion: White matter lesions impact the cognitive function in patients with lacunar infarction. The more severe the white matter lesions, the more significant decline in cognitive function.

Human mesenchymal stem cells modified by hepatocyte growth factor gene promote chicken embryonic angiogenesis / 中国实验血液学杂志

This study was purposed to investigate the angiogenesis-promoting activities of human mesenchymal stem cells (hMSCs) modified by hepatocyte growth factor (HGF) and the underlying mechanisms. The hMSCs were transfected by recombinant adenoviral vector carrying human HGF gene and seeded onto the chicken chorioallantoic membrane. Three days later, the number of blood vessels was counted and their angiogenic response was compared with those of hMSCs of same generation, recombinant basic fibroblast growth factor (bFGF) and alpha-MEM as control. The expression levels of bFGF, VEGF, angiopoietin-1 and angiopoietin-2 were evaluated by RT-PCR assay. The results showed that gene-modified hMSCs exhibited greatest activity to promote angiogenesis while the angiogenic response was nearly same between groups treated by hMSCs and bFGF, all of which were significantly higher than that observed in control (p < 0.01). RT-PCR analysis revealed that hMSCs constitutively expressed multiple angiogenesis-associated growth factors and their levels seemed up-regulated by HGF gene transfer. It is concluded that HGF gene-modified hMSCs show a potent angiogenesis-promoting function and may be useful in the treatment of ischemic disorders.

A study of CXCR4/SDF-1 in hepatocellular carcinoma and liver cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expressions of stromal cell-derived factor-1 (SDF-1) and CX chemokine receptor-4 (CXCR-4) in patients with hepatocellular carcinoma (HC) and liver cirrhosis.</p><p><b>METHODS</b>Peripheral blood and/or ascites fluid were collected from 39 hepatocellular carcinoma patients, 16 patients with liver cirrhosis, 12 with hepatitis and 12 healthy donors. The SDF-1 expression was assayed by ELISA and CXCR-4 was measured by immunohistochemical methods.</p><p><b>RESULTS</b>The level of SDF-1 expression in the carcinoma patients was higher than that of the liver cirrhosis, hepatitis patients and healthy donors, but there was no significant difference between those of the healthy donors and hepatitis patients or liver cirrhosis patients. The levels of CXCR-4 expression were closely related to the tumor differentiation.</p><p><b>CONCLUSION</b>The expression of SDF-1 in the peripheral blood and the CXCR4 expression in the HCC tissues of the HC patients may be regarded as markers of HC and they may have a positive relationship with the differentiation and metastasis of HC.</p>

Research progress in hirudin fusion protein--review / 中国实验血液学杂志

Natural hirudin extracted from the secretion of medical leech salivary gland is a single-chain peptide containing 65 aminoacid residues with molecular weight of 7000 D, and exists in three isomers of HV1, HV2 and HV3. Hirudin possesses three disulfide bridges forming the structure of core cyclic peptides, which binds to the catalytic site of thrombin so as to inhibit the catalysis of thrombin. Its c-terminus rich in acidic aminoacid residues possesses hydrophilicity, and is free on the molecular surface, and can bind with fibrin recognition site of hirudin. The minimal segment of 12 - 16 C-terminal acidic residues keeps the minimal activity of anti-thrombosis. Thus, hirudin, as a potent and specific inhibitor of thrombin, can be used to protect from and to treat clinically thrombosis. As it has some disadvantages such as short half-life, bleeding side-effect and mono-function, and so on, hirudin has been fused with some other functional proteins in recent years. The obtained fusion proteins can prolong the half life of hirudin, or relieve it bleeding side effect, or bring new functions, such as thrombolysis, inhibiting the platelet aggregation, targeting specifically. The research progress in hirudin fusion protein was summarized in this review.

Transsphenoidal microsurgery of non-adenomas disease intrasellar region:a report of 23 cases / 中华显微外科杂志

Objective To evaluate the overall effect of transsphenoidal microsuegery for 23 patients with non-adenomas disease intrasellar region.Methods A toal of 23 patients with non-adenomas disease in- trasellar region,7 cases of Rathke's cleft cyst,3 cases of craniopharyngiomas,2 cases of meningiomas,2 ca- ses of pituitary tuberculous granuloma,5 cases of pituitary abscess,2 cases of empty sella,2 cases of chord- mas,were treated via sublabio-septo-sphenoidal microsurgery.Results There were among the 23 patients, groos total removal of the disease were achieved in 14 cases,subtotal removal in 7 cases,and partial removal in remain 2 cases.Diminished visual activity and visual field defects were improved in 13 cases.Menstrual disorder in all female cases and sexual disturbance in male were improved.There was no death in group. Conclusion Microsurgical technique via transsphenoidal approach is a safe and effective one for the treatment of patients with non-adenomas disease intrasellar region.

Purification of Recombinant Fusion Protein Staphylokinase-Hirudin Expressed by Escherichia coli and Analysis of its Dimer / 中国生物工程杂志

The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

Design and Functional Investigation of a Novel Anti-coagulative Fusion Protein by Hirudin With a Recognizing Sequence of FXa / 中国生物工程杂志

Hirudin (HV) is known as the most potent and specific inhibitor of thrombin. Although hirudin has many advantages , it has the bleeding side effect and this is the great shortage of hiudin for clinical application. In order to alleviate bleeding side effect of hirudin, fusion protein, named as FHV (fusion hirudin linked with FXa recognition peptide) was designed. The fusion protein gene ( fhv) was cloned into plasmid pPIC9K. FHV engineered Pichia pastoris containing high copies was chosen for fermentation and purification at 30 L fermentor scale, finally, FHV with purity of above 97% was obtained. To investigate the function of FHV in vivo, mouse tail thrombosis model was used. In the mice thrombus tail model induced by carrageenan, FHV decreased the length of tail thrombus significantly, similar to that of HV control, and had no obvious effects on the TT, PT and APTT. In conclusion, FHV is constructed and expressed in yeast. FHV fusion proteins is obtained by fermentation and purification. FHV has antithrombotic effects not influencing IT, PT and APTT after administration immediately in animal models. Therefore, FHV is a promising anticoagulant and antithrombotic drug.