Cloning, characterization, and heterologous expression of a candidate Hirudin gene from the salivary gland transcriptome of Hirudo nipponia.

Hirudin is a pharmacologically active substance in leeches with potent blood anticoagulation properties. Although recombinant hirudin production isolated from Hirudo medicinalis Linnaeus and Hirudinaria manillensis Lesson is known, to our knowledge, this study is the first to report recombinant hirudin expression and production from Hirudo nipponia Whitman. Thus, the present study aimed to clone and characterize the full-length cDNA of a candidate hirudin gene (c16237_g1), which is localized on the salivary gland transcriptome of H. nipponia, and further evaluate its recombinant production using a eukaryotic expression system. The 489-bp cDNA possessed several properties of the hirudin "core" motifs associated with binding to the thrombin catalytic pocket. A fusion expression vector (pPIC9K-hirudin) was constructed and successfully transformed into Pichia pastoris strain GS115 via electroporation. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blot analysis confirmed hirudin expression. The recombinant protein was expressed with a yield of 6.68 mg/L culture. Mass spectrometry analysis further confirmed target protein expression. The concentration and antithrombin activity of purified hirudin were 1.67 mg/mL and 14,000 ATU/mL, respectively. These findings provide a basis for further elucidating the molecular anticoagulation mechanism of hirudin, and address China's growing market demand for engineered H. nipponia-derived hirudin and hirudin-based drugs.

Metal-Polyphenol Network-Mediated Protein Encapsulation Strategy Facilitating the Separation of Proteins and Metabolites in Biospecimens.

Access to both protein and metabolite biomarker information in biospecimens from trace samples remains a significant challenge, and it is necessary to separate proteins and metabolites before analysis. In this work, the Fe3O4@SiO2@Proteins@Metal-polyphenol network (MPN) was successfully constructed and applied to separate metabolites and proteins. Tannic acid (TA) and Cu2+ were involved in the synthesis of MPN because of rapid degradation and maintaining the assay performance of proteins. There are a variety of interactions between TA and proteins, including hydrogen-bonding, hydrophobic, and ionic interactions. Moreover, benefiting from the small molecule permeability and surface adherence of MPN, proteins were encapsulated and immobilized on the surface of substrates with the growth of MPN. At the same time, endogenous metabolites remained dispersed in the supernatant. In the model sample and real biospecimen cases, the protein biomarkers (e.g., carcinoembryonic antigen and alanine aminotransferase) were encapsulated on the surface of Fe3O4@SiO2, which allowed the isolation of proteins from the original matrix, as well as release and analysis in a short time. Meanwhile, the metabolites in the produced supernatant were analyzed by LC-MS/MS. By the self-assembly and disassembly of MPN, the group differences of proteins and metabolites between physiological and pathological biospecimens are correctly characterized without multisampling. Overall, an MPN-mediated separation strategy of biomarkers was proposed, and MPN facilitated a "two birds with one stone" approach, where the proteins were encapsulated and immobilized in the precipitation while endogenous metabolites distributed in the produced supernatant, opening a new chapter in the application of MPNs.

A comparative study of Liandan Xiaoyan Formula metabolic profiles in control and colitis rats by UPLC-Q-TOF-MS combined with chemometrics.

Liandan Xiaoyan Formula (LDXYF) is a traditional Chinese medicine prescription (TCMP) consisting of Herba Andrographis (dried herb of Andrographis paniculata) and Picrasmae ramulus et folium (dried twiggeries and leaves of Picrasma quassioides). It is used to treat diarrhea, acute gastroenteritis, colitis, and dysentery, among other inflammatory gastrointestinal diseases. However, because of less research on the in vitro chemical composition and holistic metabolism of LDXYF, in vivo mechanisms of action and quality control of LDXYF have not yet been fully assessed due to the lack of studies into its bioactive components. In this study, ultra-performance liquid chromatography coupled with quadruple time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was established for comprehensive analysis of chemical compounds of LDXYF and their metabolites in serum and urine samples of control and colitis rats. As a result, totally 94 compounds in LDXYF were unambiguously identified or tentatively characterized. And a total of 91 LDXYF-related xenobiotics were characterized, including 61 (16 prototypes and 45 metabolites) in serum, and 72 (26 prototypes and 46 metabolites) in urine. Besides, we compared the exposure of metabolites in normal and colitis rats by chemometrics and summarize similarities and differences of metabolic pathways of mainly compounds in normal and colitis conditions, and found that in control and colitis conditions, alkaloids predominantly went through phase I reaction combined phase II reaction (hydroxylation and sulfation, hydroxylation and glucuronidation, demethylation and glucuronidation), while the major metabolic reaction of diterpene lactones were phase Ⅱ reactions (glucuronidation, sulfation). And there were no significant differences in metabolic pathways between control and colitis groups, just the exposure of prototype and their metabolites absorbed into serum or excreted through the urine were different, and 17 alkaloids and 6 diterpene lactone prototypes and their metabolites in serum could be considered as potential pharmacodynamic substances. A comprehensive analysis of the compounds and metabolic characteristics of LDXYF was conducted in our study, and the results laid the chemical foundation for further research into effective substances and the action mechanism of LDXYF.

One-stage anterior focus debridement, interbody bone graft, and anterior instrumentation and fusion in the treatment of short segment TB.

To evaluate the clinical efficacy of 1-stage anterior focus debridement, interbody bone graft, and anterior instrumentation and fusion in the treatment of short segment thoracic tuberculosis with paraplegia or incomplete paralysis. A total of 16 adult patients with short segment thoracic spinal thoracic tuberculosis who underwent surgery were enrolled in this retrospective study. All patients received anterior focus debridement, interbody bone graft and anterior instrumentation and fusion. All patients were followed up for 24 to 48 months. Clinical manifestations, laboratory examinations, neurological symptoms, bone fusion and imaging results were analyzed. All patients successfully underwent operations. The symptoms of chest and back pain were alleviated and even disappeared during postoperative 1 to 6 months. There was no recurrence. All patients got bony spinal fusion within postoperative 4 to 8 months assessed by spinal X-ray film. The levels of erythrocyte sedimentation rate and C-reactive protein were significantly decreased from 72.6 ±â€…27.5 mm/h and 75.7 ±â€…25.9 mg/L to 15.9 ±â€…4.6mm/h and 4.7 ±â€…2.0mg/L at the final follow-up, respectively (P < .05). The thoracic kyphosis angle was also notably decreased from 15.0 ±â€…3.4° to 9.1 ±â€…1.9° after operation(P < .05). During the follow-up, the symptom of paraplegia or incomplete paralysis was significantly improved. Neurologic status in all patients was also improved to some extent. The combination of 1-stage anterior focus debridement, interbody bone graft and anterior instrumentation and fusion is an effective and feasible treatment method for short segmental thoracic tuberculosis with paraplegia or incomplete paralysis.

One-step fabrication of COF-coated melamine sponge for in-syringe solid-phase extraction of active ingredients from traditional Chinese medicine in serum samples.

In this study, a covalent organic framework (COF)-TpBD-supported melamine sponge (MS) was fabricated through a one-step hydrothermal method. The obtained monolithic column was then applied in in-syringe solid-phase extraction (IS-SPE) for the separation of three volatile ingredients from serum samples. Given credit for the superior adsorption capacity of the COF and the homogeneous microporous property of MS, the developed column exhibited satisfactory separation of the targets. And the dominating adsorption mechanism was the hydrophobic interaction forces between TpBD and targets and the high mass transfer efficiency provided by the large pore structure of MS. The results of dynamic adsorption showed that the MS@TpBD column displayed much better adsorption performance than blank MS and TpBD. And it has featured great reusability up to 5 cycles and obtained satisfied recovery values (87.9 ~ 110.3%) in serum samples. As a result of sample clean-up, this column offers low limit of detections (LODs) down to 0.014, 0.010, and 0.020 µg/mL, respectively. In summary, we believe that this convenient separation column has prominent application promise in the fields of separating activity ingredients in biological samples.

The Effect of Fine-Grained Control of Workflow on Informal Learning: The Mediating Effect of Teacher Burnout.

Workflow fine control is an important research tool for the mediating effect of teacher burnout. Based on the fine control theory of workflow, this paper constructs a mediating effect model of teacher burnout in primary and secondary school teachers from the perspective of informal learning. The model designs a measurement scale based on the results of fine control, and the scale is revised through informal learning pretests to form the logical frame analysis loop of this paper. The scale uses Likert 5-point scoring, from 1.5 to represent never, rarely, occasionally, often, and always, which solves the difficulty of quantitative analysis of teacher burnout data. During the simulation process, SPSS 20.0 and AMOS 7.0 were used to conduct empirical analysis, exploratory factor analysis and confirmatory factor analysis on the survey data, and the measurement scales were tested. The empirical analysis results show that this paper uses cluster sampling and random sampling to carry out a questionnaire survey on 1022 primary and secondary school teachers. The measurement scale includes 19 items. Through exploratory factor analysis and confirmatory factor analysis, the control method is determined. In two dimensions, the revised control style questionnaire has a reliability of 0.951, and items with factor loadings less than 0.5 are eliminated, which effectively improves the control performance of informal learning.

Pharmacokinetic Study of Four Major Bioactive Components of Liandan Xiaoyan Formula in Ulcerative Colitis and Control Rats Using UPLC-MS/MS.

Liandan Xiaoyan Formula (LXF), a classic Traditional Chinese medicine (TCM) formula, is composed of two Chinese herbal medicines for treating bowel disease under the TCM theory. This study aimed to develop a rapid, stable, sensitive, and reliable method based on ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to simultaneously determine four major bioactive components of LXF (andrographolide, dehydroandrographolide, 1-methoxicabony-ß-carboline, 4-methoxy-5-hydroxy-canthin-6-one) in rat serum and evaluate the pharmacokinetic characteristics of LXF in ulcerative colitis (UC) and control rats. After pretreating by protein precipitation with methanol, separation was performed on a UPLC C18 column using gradient elution with a mobile phase consisting of acetonitrile and 0.1% formic acid at a flowing rate of 0.4 ml/min. Detection was performed on Triple-TOF™ 5600 mass spectrometry set at the positive ionization and multiple reaction monitoring (MRM) mode. The validated method showed good linearity (R 2 ≥ 0.9970), the intra- and inter-day accuracy were within ±11.58%, whereas the intra- and inter-day precision were less than 13.79%. This method was validated and applied to compare the pharmacokinetic profiles of the analytes in serum of UC induced by dextran sulphate sodium (DSS) and control rats after oral administration of LXF. The results showed that four major bioactive components of LXF were quickly absorbed after oral administration in both groups, with higher exposure levels in the UC group. This relationship between the active ingredients' pharmacokinetic properties provided essential scientific information for applying LXF in clinical.

In situ self-assembly of three-dimensional porous graphene film on zinc fiber for solid-phase microextraction of polychlorinated biphenyls.

Three-dimensional porous graphene film (3DPGF) was fabricated on Zn fiber for solid-phase microextraction (SPME) through in-situ self-assembly strategy at room temperature. Fast electron transfer due to ionization of zinc induces reduction of graphene oxide and thus leads to the layer-by-layer interfacial deposition of graphene sheets, forming three-dimensional porous network morphology. The 3D interpenetrating porous structure could provide more available adsorption site for the target molecules and also enhance mass transfer efficiency in the extraction process. Therefore, the obtained 3DPGF fiber exhibited excellent performance when applied in the SPME of preconcentration and quantification of polychlorinated biphenyls (PCBs) in environment waters. The developed method showed a linear range from 1.0 ng L-1 and 200 ng L-1 with an acceptable correlation (R2=0.990). The limit of detection (LOD) and limit of quantification (LOQ) were found 0.03-0.2 ng L-1 and 0.1-0.8 ng L-1, respectively. The proposed method was applied in real water samples analysis with the recoveries ranging from 63.1 to 111.3%. The present study expanded the application of three-dimensional porous graphene materials in sample preparation and revealed its potential in SPME application.

Effects of Nervilia fordii Extract on Pulmonary Fibrosis Through TGF-ß/Smad Signaling Pathway.

Idiopathic pulmonary fibrosis (IPF) is a progressive and irreversible interstitial pulmonary disease with a poor prognosis. The extract of Nervilia fordii (NFE) has shown remarkable benefit in the treatment of acute lung injury, lung cancer, and severe acute respiratory syndrome (SARS). However, the potential mechanism and efficacy of NFE in the treatment of IPF remain unknown. In this study, a systematic network pharmacology analysis was used to predict the mechanism and efficacy of NFE in the treatment of IPF, based on the major components of NFE elucidated by UPLC-TOF-MS/MS. The potential molecular interactions between the compounds and potential targets were predicted using molecular docking. In vivo, rats with pulmonary fibrosis induced by a single intratracheal injection of bleomycin (BLM) were orally administered NFE for 14 days. Lung index and biochemical levels were determined, and histopathological analysis using hematoxylin and eosin (H&E) and Masson staining was performed. The effects of NFE on fibroblast proliferation in Lipopolysaccharide (LPS) and TGF-ß1-induced mouse 3T6 fibroblasts were evaluated in vitro. In total, 20 components were identified in NFE, and 102 potential targets for IPF treatment were predicted. These targets potentially participate in processes regulated by transmembrane receptor protein tyrosine kinase, ERBB2, and et al. Molecular docking results predicted high affinity interactions between three components (rhamnazin, rhamnetin, and rhamnocitrin) and the potential targets, suggesting that TGF-ß is the most important potential target of NFE in the treatment of pulmonary fibrosis. NFE significantly decreased the lung index and alleviated BLM-induced pulmonary fibrosis in rats. Histopathological observation of lung tissues showed that NFE alleviated inflammation and collagen deposition in BLM-induced rats. NFE inhibited the migration of LPS- and TGF-ß1-induced 3T6 fibroblasts, reduced the contents of hydroxyproline and collagen, and contributed to anti-inflammation and anti-oxidation. With the intervention of NFE, the protein and RNA expression of TGF-ß1, a-SMA, Smad3/4, p-Smad3/4, CTGF, and p-ERK1/2 were significantly downregulated, while Smad7 and ERK1/2 were upregulated significantly in vivo and in vitro. These findings indicated that NFE may exert therapeutic effects on pulmonary fibrosis by alleviating inflammation, oxidation, and collagen deposition. The mechanism related to the inhibition of the TGF-ß/Smad signaling pathway.

Network pharmacology-based mechanism prediction and pharmacological validation of Xiaoyan Lidan formula on attenuating alpha-naphthylisothiocyanate induced cholestatic hepatic injury in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The well-known Chinese prescription, Xiaoyan Lidan Formula (XYLDF), possesses efficiency of heat-clearing, dampness-eliminating and jaundice-removing. It has long been used clinically for the treatment of hepatobiliary diseases due to intrahepatic cholestasis (IHC). However, the mechanism of XYLDF for its therapeutic effects remains elusive. AIM OF THE STUDY: The study aimed to explore the potential targets for liver protective mechanism of XYLDF based on network pharmacology and experimental assays in ANIT-induced cholestatic hepatic injury (CHI) in rats. MATERIALS AND METHODS: On the basis of the 29 serum migrant compounds of XYLDF elucidated by UPLC-TOF-MS/MS, a network pharmacology approach was applied for the mechanism prediction. Systematic networks were constructed to identify potential molecular targets, biological processes, and signaling pathways. And the interactions between significantly potential targets and active compounds were simulated by molecular docking. For the mechanism validation, an ANIT-induced rat model was used to evaluate the effects of XYLDF on CHI according to serum biochemistry, bile flow rates, histopathological examination, and the gene and protein expression including enzymes related to synthesis, export, and import of bile acid in liver and ileum, and those of inflammatory cytokines, analyzed by RT-qPCR and WB. RESULTS: The results of network pharmacology research indicated TNF (TNF-α), RELA (NF-κB), NR1H4 (FXR), and ICAM1 (ICAM-1) to be the important potential targets of XYLDF for cholestatic liver injury, which are related to bile metabolism and NF-κB-mediated inflammatory signaling. And the molecular docking had pre-validated the prediction of network pharmacology, as the core active compounds of XYLDF had shown strong simulation binding affinity with FXR, followed by NF-κB, TNF-α, and ICAM-1. Meanwhile, the effects of XYLDF after oral administration on ANIT-induced CHI in rats exhibited the decreased levels of transaminases (ALT and AST), TBA, and TBIL in serum, raised bile flow rates, and markedly improved hepatic histopathology. Furthermore, consistent to the above targets prediction and molecular docking, XYLDF significantly up-regulated the expression of FXR, SHP, BSEP, and MRP2, and down-regulated CYP7A1 and NTCP in liver, and promoted expression of IBABP and OSTα/ß in ileum, suggesting the activation of FXR-mediated pathway referring to bile acid synthesis, transportation, and reabsorption. Moreover, the lower levels of TNF-α in plasma and liver, as well as the reduced hepatic gene and protein expression of NF-κB, TNF-α, and ICAM-1 after XYLDF treatment revealed the suppression of NF-κB-mediated inflammatory signaling pathway, as evidenced by the inhibition of nuclear translocation of NF-κB. CONCLUSIONS: XYLDF exhibited an ameliorative liver protective effect on ANIT-induced cholestatic hepatic injury. The present study has confirmed its mechanism as activating the FXR-regulated bile acid pathway and inhibiting inflammation via the NF-κB signaling pathway.

5-Hydroxy-4-methoxycanthin-6-one alleviates dextran sodium sulfate-induced colitis in rats via regulation of metabolic profiling and suppression of NF-κB/p65 signaling pathway.

BACKGROUND: 5-Hydroxy-4-methoxycanthin-6-one (PQ-A) is the main active compound in Ramulus et Folium Picrasmae, a Chinese herbal medicine commonly used in colitis treatment. PURPOSE: To clarify PQ-A's role and mechanism in colitis treatment based on a non-targeted metabolomics study. METHODS: Rats with ulcerative colitis (UC) established with 4% dextran sulfate sodium (DSS) were orally treated with PQ-A. Body weight, disease activity index (DAI), colon length, biochemical parameters (MDA and SOD), and histopathological score in colon tissue were measured. A UPLC-Q-TOF-MS/MS approach-based metabolomics analysis was conducted to explore the underlying mechanisms of PQ-A in colitis treatment. Inflammatory cytokines (TNF-α, IL-1ß, IL-6, and IL-10) concentrations in serum and their protein levels in the colon were determined. CD3 and NF-κB/p65 immunohistochemistry in the colon was semi-quantified. The related protein or mRNA in IKK-NF-κB/p65 signaling pathway was measured by Western blotting or RT-PCR, respectively. Potential molecular interactions between PQ-A and NF-κB/p65 was predicted using DS 2.5 software. RESULTS: PQ-A significantly prevented body weight loss and colonic shortening in colitic rats, and reduced the DAI and histopathologic score as well. PQ-A decreased MDA levels in the UC rat serum and increased those of SOD. Metabolomics results revealed forty-nine differential metabolites as biomarkers of DSS-induced colitis, demonstrating that the path-mechanism of colitis involved the perturbation of eight metabolic pathways, including alpha-linolenic acid and linoleic acid metabolism, sphingolipid metabolism, retinol metabolism, bile acid metabolism, et al. Thirty-six biomarkers were especially reversed to normal-like levels by PQ-A via regulation of alpha-linolenic acid and linoleic acid metabolism, sphingolipid metabolism, and retinol metabolism, which effectively hinted the potential pharmacological mechanism of PQ-A related to NF-κB/p65 inflammatory signaling. Molecular docking results predicted high affinity interaction between PQ-A and NF-κB/p65, involving hydrogen-bond interactions at five amino acid residues, suggesting NF-κB/p65 as a target. PQ-A decreased TNF-α, IL-1ß, and IL-6 concentrations in serum and their protein levels in colon tissue in colitic rats. CD3, MYD88, p-IκBα, NF-κB/p65, and p-NF-κB/p65 expression levels decreased, whereas those of IKKß and IκBα increased in colitic tissue following PQ-A treatment. PQ-A strongly inhibited nuclear translocation of NF-κB/p65. CONCLUSIONS: We provide an overview of PQ-A's possible mechanism of action in colitis treatment based on serum non-targeted metabolomics. PQ-A treatment can protect rats against DSS-induced colitis by suppressing the NF-κB/p65 signaling pathway.

TMPO-AS1, a Novel E2F1-Regulated lncRNA, Contributes to the Proliferation of Lung Adenocarcinoma Cells via Modulating miR-326/SOX12 Axis.

BACKGROUND: TMPO-AS1, an antisense lncRNA located at human chromosome 12p23.1, has been identified as an oncogene involved in cell proliferation in various cancers, including LUAD. In this study, we aimed to explore the novel molecular mechanism of TMPO-AS1 underlying LUAD growth. MATERIALS AND METHODS: The transcription levels of TMPO-AS1, miR-326, and SOX12 in LUAD tissues and cell lines were detected by quantitative real-time PCR (qRT-PCR). The cell proliferation ability was evaluatect 3d by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis analysis was assessed by flow cytometry. The target relationship among TMPO-AS1, miR-326, and SOX12 and promoter activity of TMPO-AS1 was measured using dual-luciferase reporter assay. The protein levels of SOX12 in LUAD cells were determined by Western blot. ChIP-qPCR assay was performed to validate the direct binding between E2F1 and TMPO-AS1 promoter. RESULTS: TMPO-AS1 was up-regulated in LUAD tissues as well as cell lines. Boosted TMPO-AS1 expression was positively correlated with poor prognosis and pathological stage in LUAD. Down-regulation of TMPO-AS1 could restrain the proliferation of LUAD cells through arresting the cell cycle at G0/G1 phase and inducing apoptosis in vitro. Mechanically, we demonstrated that TMPO-AS1 could modulate the proliferation of LUAD cells through increasing SOX12 expression level via sponging miR-326 in accordance with bioinformatics analysis and experimental validation. Furthermore, we identified that TMPO-AS1 could be activated by E2F transcription factor 1 (E2F1) as a novel target gene. CONCLUSION: TMPO-AS1 can modulate LUAD cell proliferation through E2F1/miR-326/SOX12 pathway.

Comparative study of the effect of rice husk-based powders used as physical conditioners on sludge dewatering.

The effects of rice husk flour (RHF), rice husk biochar (RHB), and rice husk-sludge cake biochar (RH-SCB, expresses sludge cake biochar deriving from a sludge that has been previously conditioned with rice husk) used as physical conditioners on sludge dewaterability were compared. The effects of characteristics of physical conditioners on sludge compressibility and zeta potential were analyzed. The optimal rice husk-based powder was RH-SCB, which presented the highest net sludge solid yield (YN, expresses the dry mass flow by filtration) at 20.39 kg/(m2 h) for 70% dry sludge (DS). Characterization analysis indicates that the hardness and surface Fe content of powders which could influence the compressibility coefficient of sludge cake and sludge zeta potential were the major factors influencing sludge dewaterability. The comparison of feasibility and economic analysis showed that adding RH-SCB improves the quality of the sludge filtrate and reduces the pollution potential of conditioned sludge (the ratio of secondary and primary (RSP) of Cu, Zn, Cd reduces from 43.05, 144.00, 7.25 to 7.89, 14.63, 4.27, respectively), and the costs of using RH-SCB were the lowest (at 88.4% lower than that of the raw sludge). Therefore, it is feasible to use RH-SCB to improve sludge dewaterability.

Photocatalytic 1,1-Hydrofluoroalkylation of Alkynes with a Concurrent Vicinal Acylation: An Access to Fluoroalkylated Cyclic Ketones.

A visible-light-induced 1,1-hydrofluoroalkylation of alkynes with a concomitant vicinal acylation is developed using tetrahydrofuran (THF) as the hydrogen atom source. Various fluoroalkylated cyclic ketones, such as indanones, chroman-4-ones, 2,3-dihydroquinolin-4(1 H)-ones, and 3,4-dihydronaphthalen-1(2 H)-ones, can be efficiently synthesized with excellent trans-diastereoselectivity. The reaction represents the first example of 1,1-hydrofluoroalkylation of alkynes, thus providing a novel method for the construction of fluoroalkanes.

Transcriptomic analysis of the salivary gland of medicinal leech Hirudo nipponia.

Hirudo nipponia (known as Shui Zhi in Chinese) is a well-known Chinese medicine with numerous active ingredients in its body, especially in its saliva. This native Chinese blood-sucking leech has been used for therapeutic purposes since before 100 AD. Modern Chinese physicians use it for a wide range of diseases. Genomic data and molecular information about the pharmacologically active substances produced by this medicinal leech are presently unavailable despite this organism's medicinal importance. In this study, we performed transcriptome profiling of the salivary glands of medicinal leech H. nipponia using the Illumina platform. In total, 84,657,362 clean reads were assembled into 50,535 unigenes. The obtained unigenes were compared to public databases. Furthermore, a unigene sequence similarity search and comparisons with the whole transcriptome of medical leech were performed to identify potential proteins. Finally, more than 21 genes were predicted to be involved in anticoagulatory, antithrombotic, antibacterial, anti-inflammatory and antitumor processes, which might play important roles in the treatment of various diseases. This study is the first analysis of a sialotranscriptome in H. nipponia. The transcriptome profile will shed light on its genetic background and provide a useful tool to deepen our understanding of the medical value of H. nipponia.

Complete mitochondrial genome of Hepialus xiaojinensis (Lepidoptera: Hepialidae).

Although Hepialus xiaojinensis is important as the host of Ophiocordyceps sinensis, it remains poorly known especially in genetic structure and phylogeny status. To get a better understanding of it, the complete mitochondrial genome sequence of H. xiaojinensis was sequenced for the first time. The genome was 15 397 bp in size and contained 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and an AT-rich region, and the gene composition and the arrangement of which were consistent with other insects of Hepialidae. The base composition of the genome was A (41.44%), T (39.68%), C (11.52%), and G (7.36%), with an AT content of 81.13%. Moreover, we have performed phylogenetic analysis of H. xiaojinensis and other 11 Lepidoptera species.

Atypical imaging of spinal tuberculosis: a case report and review of literature.

This is a case report of spinal tuberculosis combined with sacroiliac joint tuberculosis, pulmonary tuberculosis, chest wall tuberculosis and tuberculous pleurisy and the image of the patient is rare, special and not typical and it looks like a halo sign. It has an important reference value for the diagnosis of spine tuberculosis although it is a rare imaging manifestation and diagnosis was confirmed by pathology after the surgery. Therefore atypical imaging is often appeared in clinical practice and it is meaningful and necessary for the diagnosis of atypical spinal tuberculosis combined with multiple organ tuberculosis.

[Histological Study on Integumentary and Digestive System of Hirudo nipponica].

Objective: The integumentary and digestive system of Hirudo nipponica were studied,in order to protect the precious Chinese herbal medicine. Methods: The histological study of integumentary and digestive system of Hirudo nipponica by histological methods were carried out by microscopical method. Results: The body wall of Hirudo nipponica could be divided into five layers, including cuticular layer, epithelial layer, dermis layer, muscle layer and grape-like tissue. The digestive system was composed ofmouth,pharyngeal,esophagus,crop,intestine,rectum and anus. Conclusion: This study can provide the basic data for the artificial breeding, research in pathology and physiological functions of Hirudo nipponica.