Four small GTPase genes which may be relative to sclerotial development were firstly cloned from medicinal fungus Polyporus umbellatus using rapid amplification of cDNA end PCR (RACE) method. The results showed that full-length cDNA of PuRhoA was 698 bp contained 585 bp ORF, which was predicted to encode a 194 amino acid protein with a molecular weight of 21.75 kD with an isoelectric point (pI) of 6.44; the full length cDNA of PuRhoA2 was 837 bp in length and encoded a 194 amino acid protein with a molecular weight of 21.75 kD and an isoelectric point (pI) of 6.33; the full length cDNA of Puypt1 was 896 bp in length and encoded a 204-aa protein with a molecular weight of 22.556 kD and an isoelectric point (pI) of 5.75; the full length cDNA of PuRas was 803 bp in length and encoded a 212-aa protein with a molecular weight of 23.821 kD and an isoelectric point (pI) of 5.2. There are fani acyl transferase enzyme catalytic site and myrcene-transferase enzyme catalytic site in PuRhoA1 while the PuRhoA2 only possess myrcene-transferase enzyme catalytic site. Puypt1 contains the Rab1-Ypt1 conserved domain of small GTPase family and PuRas contains the fani acyl transferase enzyme catalytic site. According to the phylogenetic analysis all these four small GTPase clustered with basidiomycete group. Quantitative real-time PCR analysis revealed that Puypt1, PuRas and PuRhoA1 transcripts were significantly higher in the beginning of sclerotial formation than that in the mycelia, whereas the transcripts levels of PuRhoA2 gene were particularly lower in sclerotia than that in mycelia, suggesting that these four genes might be involved in P umbellatus selerotial development.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Fungal Proteins ; genetics ; GTP Phosphohydrolases ; genetics ; Genes, Fungal ; Mycelium ; Phylogeny ; Polyporus ; enzymology ; genetics ; Real-Time Polymerase Chain Reaction

To investigate such physical indexes as hygroscopicity, angle of repose, bulk density, fillibility of compression of mixed powder of directly compressed auxiliary materials and fermented cordyceps powder by using micromeritic study methods. The results showed that spray-dried lactose Flowlac100 and microcrystalline cellulose Avicel PH102 had better effect in liquidity and compressibility on fermented cordyceps powder than pregelatinized starch. The study on the impact of directly compressed auxiliary materials on the powder property of fermented cordyceps powder had guiding significant to the research of fermented cordyceps powder tablets, and could provide basis for the development of fermented cordyceps powder tablets.
Cellulose ; chemistry ; Cordyceps ; chemistry ; Drug Compounding ; methods ; Fermentation ; Lactose ; chemistry ; Powders ; chemistry ; Tablets ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate effects of the variation of immune function in high humidity environment in different time, and lay a foundation for further study of the related mechanism.

METHODThirty SD rats were divided into 3 groups (n = 10): 20 day group, 40 day group in 90% relative humidity chamber and control group in normal relative humidity. Peripheral blood and spleens were collected to detect the levels of T lymphocyte subsets by Flow Cytometery.

RESULTSIn peripheral blood of the 20 day group rats, the CD3+ %, CD4+ %, CD8+ % and CD4+/CD8+ were 52.91 +/- 6.27, 37.80 +/- 4.11, 14.85 +/- 3.73 and 2.72 +/- 0.82 separately. Expect CD3+ %, they all had significant differences (P < 0.05). In addition, the data of the 40 day group rats showed no diversity in statistics. In spleen, CD8+ % of the 20 day group rats was 6.23 +/- 2.87 with significant differences (P < 0.05) and IgG, IgA and IgM did not change a lot in blood serum of the high humidity groups except C3 of the 20 days group (P < 0.05).

CONCLUSIONIn high humidity environment, the immune function of the rats increased in the initial stage. As time went on, the immune function gradually went to normal level through the self adjustment.

Acclimatization ; Animals ; Humidity ; Rats ; Rats, Sprague-Dawley ; Spleen ; immunology ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo compare the clinical curative effect of thoracolumbar burst fracture treated by the posterior unilateral approach corpectomy fusion screw-rod fixation and anterior corpectomy bone fusion screw plate fixation.

METHODSFrom January 2008 to May 2014,36 cases of thoracolumbar burst fracture underwent operation of decompression, fusion, and internal fixation was retrospective analyzed. Among them, 16 patients were treated through posterior approach as posterior group, including 13 males and 3 females aged from 37 to 62 years old; 9 cases caused by falling injury, 3 cases by traffic accident injury,4 cases by heavy aboved;the injury segment was on T₁₂ in 2 cases, L₁ in 5 cases, L₂ in 7 cases, L₃ in 2 cases; according ASIA grade, 3 cases were grade A, 2 cases were grade B, 2 cases were grade C, 5 cases were grade D, 4 cases were grade E; the time between injury and operation ranged from 5 to 15 days. Other 20 patients were treated through anterior-lateral approach as anterior-lateral group, including 15 males and 5 females with age from 27 to 62 years old; 12 cases caused by falling injury, 4 cases by traffic accident injury, 4 cases by heavy aboved; the injury segment was on T₁₂ in 2 cases, L₁, in 7 cases, L₂ in 9 cases, L₃ in 2 cases; for ASIA grade: 4 cases were grade A, 2 cases were grade B, 4 cases were grade C, 6 cases were grade D, 4 cases were grade E; the time between injury and operation ranged from 4 to 12 days. The operation time, bleeding during operation and postoperative drainage volume were observed in two groups,and the changes of nerve function of ASIA grade, clinical efficacy,improved degree of thoracic and lumbar lordosis,and bony fusion were compared between two groups.

RESULTSAll patients were followed up from 12 to 24 months with an average of (15.8 ± 3.3) months. The operation time, bleeding during operation, and postoperative drainage volume had no significant different between two groups (P > 0.05). As compared with preoperative, ASIA grade of two groups at last follow-up had statistically significantly different (P < 0.01), the neural function of two groups after operation was recovered for different extent. The JOA score of two groups was compared between last follow-up and preoperative, the difference had statistically significant (P < 0.01), the two groups showed good clinical effect. The clinical results of ASIA grade, JOA score and RIS had no significant differences between two groups. All patients of two groups were obtained fusion. Thoracic and lumbar lordosis angle improvement degree had no significant difference between two groups ,it bad significant difference had statistical significance compared with preoperative, the two approaches could effectively restore the spinal sequence.

CONCLUSIONFor patients with thoracolumbar burst fracture just treated by anterior decompression and reconstruction of anterior column, according to the degree of operation performer' skill proficiency and the patient' condition to choose, but for patients must performed the spinal canal decompression anterior and posterior, the three column-reconstruction to required anterior-posterior approach, the posterior unilateral approach corpectomy fusion screw-rod fixation obviously shorten operation time, reduce the operation wound, it is worth the clinical promotion.

Adult ; Decompression, Surgical ; methods ; Female ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Reconstructive Surgical Procedures ; Retrospective Studies ; Spinal Fractures ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; injuries ; surgery

Silkworm is a holometabolous insect of Lepidoptera. During metamorphosis, significant morphological changes happen including the dissociation of old tissues and remodeling of new tissues. It has been reported that cathepsins are involved in these processes. Cathepsin is a kind of intracellular proteinase that exists in many species. It includes some subfamilies like cathepsin B, H and L. The studies on cathepsin are useful for clarifying the details of silkworm metamorphosis process. In total, 13 cathepsins were identified by screening the silkworm genome database. The basic information and the expression patterns about these genes were analyzed. Interestingly, an ovary-specific cathepsin L gene (Gene ID: BGIBMGAOO4622) was investigated by the data of silkworm microarray and real-time quantitative PCR (qPCR). The full-length cDNA is 1,209 bp, encoding a protein with 402 amino acids. Sequences alignment revealed that it has a high sequence similarity with cathepsin L of other species, and it is highly conserved in the active-site of the enzyme. The phylogenetic analysis showed that ovary-specific cathepsin L is clustered with other lepidopterous insects. Furthermore, this gene was cloned and prokaryotic expressed. Recombinant protein was present in inclusion body. Importantly, the qPCR result showed that the expression level of this gene is increasing during the early stage of pupal development and reaches the highest value at the 3rd day of pupal stage, which suggests that this gene may be involved in the process of development of the ovary and oocyte.
Animals ; Bombyx ; genetics ; Cathepsins ; genetics ; Insect Proteins ; genetics ; Phylogeny

OBJECTIVE: To investigate the expression and significance of miRNA-324-3p and its target gene WNT2B in tissue specimens of nasopharyngeal carcinoma (NPC) specimens. METHOD: qRT-PCR was used to detect the expression of miRNA-324-3p and WNT2B mRNA, and Western blot was applied to assay the expression of WNT2B protein in 39 cases of NPC specimens and 21 cases of non-carcinoma epithelium. The relationship between their expression levels and clinicopathological characteristics and their correlation with clinical pathological parameters was analyzed. RESULT: The expression of miRNA-324-3p was significantly down-regulated decreased but WNT2B mRNA/protein increased obviously in NPC specimens (P < 0.01). A negative correlation between miRNA-324-3p and WNT2B was spotted (P < 0.05). The expression levels of these markers were closely correlated with T stage, clinic stage and cervical lymph node metastasis (P < 0.05). CONCLUSION The loss of miRNA-324-3p and ectopic WNT2B might co-induce the initiation and progression of NPC.
Carcinoma ; Glycoproteins ; genetics ; metabolism ; Humans ; Lymphatic Metastasis ; MicroRNAs ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; RNA, Messenger ; metabolism ; Wnt Proteins ; genetics ; metabolism

Objective To analyze the short-term clinical efficacy of the treatment of ipsilateral femoral intertrochanteric fractures with femoral head necrosis through total hip arthroplasty combined with internal fixation.Methods From January 2011 to June 2014 a total of 28 cases of ipsilateral femoral intertrochanteric fractures with femoral head necrosis were selected,and they were all treated with open reduction and greater trochanter plate fixation combined with total hip arthroplasty.The time of operation, amount of bleeding,time of bone healing of the greater trochanteric fracture,postoperative complications, rate of internal fixation device loosening,incidence of infection,and the Harris score of the hip joint before and after operation were recorded and statistically analyzed.Results The mean operative time was (174.0±27.1)min and the mean blood loss was (530.0±132.0)mL.There were 2 cases of deep venous thrombosis in the lower extremities,and there was no serious complications occured among the remaining cases.All patients were followed up for 12 to 24 months,averagely (16.3±7.0) months.All the 28 patients get bony union of the large trochanter,with the average healing time of (5.0±1.5) months.The preoperative Harris score was (28.0±7.1) points, while it was (86.5±5.3) points after the surgery, and the difference was statistically significant(t=-21.699,P=0.000).At the end of the follow-up,there was no internal fixation device loosening, no cable breakage,and no infection cases.Conclusion Total hip arthroplasty combined with internal fixation for the treatment of ipsilateral femoral intertrochanteric fractures with femoral head necrosis, especially the large trochanteric fractures, were of reliable fracture fixation, good initial stability and good joint function recovery.And it can maintain fracture stability and early recovery of joint function,so that intertrochanteric fractures and femoral head necrosis were treated simultaneously,and its recent clinical results were satisfactory.

Objective To investigate the mRNA level of glucose-6-phosphate isomerase(GPI)ex- pression in patients with rheumatoid arthritis(RA).Method Reverse transcription-polymerase chain reaction (RT-PCR)was applied to semiquantitatively analyze GPI mRNA expression in the peripheral blood mononu- clear cells(PBMCs)of 30 active RA patients,30 stable RA patients,30 patients with other rheumatic disease and 30 healthy subjects.Results There was statistically significant differences between patients with RA and other rheumatic diseases(P

OBJECTIVETo investigate the regulatory effect of potassium channel blocker (tetraethylammonium [TEA], aminopyridine [4-AP], glibenclamide [Glib]) on the proliferation of SD rat prostatic epithelial cells in vitro.

METHODSThe primary culture was prepared by collagenase dissociation of minced prostatic tissues. Cells were cultured in serum-free prostate epithelial cell growth media and identified by immunocytochemical studies. TEA and 4-AP at the concentration of 1, 5 and 10 mmol/L and Glib at the concentration of 10, 50 and 100 mol/L were added, and after 24, 48 and 72 hours of culturing, a cell column diagram was drawn and the cell number counted. The post-passage cell growth was observed by MTT assay and Hoechst33258 nucleus staining.

RESULTSThe cultured cells showed the typical morphological features of epithelia, with positive stain. MTT assay and Hoechst33258 staining showed that TEA, 4-AP and Glib at the increasing concentration effected different degrees of proliferation of prostatic epithelial cells after 24, 48 and 72 h (P < 0.01).

CONCLUSIONThe potassium channel blocker is a direct physiological regulator of the proliferation of SD rat prostatic epithelial cells.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Epithelial Cells ; drug effects ; Male ; Potassium Channel Blockers ; pharmacology ; Prostate ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo probe the function of interleukin-17 (IL-17) in rheumatoid arthritis (RA) patients of accumulated dampness-heat obstruction in joints syndrome (ADOJS) by detecting levels of IL-17 in serum and the synovial fluid and analyzing its correlation with erythrocyte sedimentation rate (ESR) and C reactive protein (CRP).

METHODSFrom January 2011 to January 2013, recruited were 90 RA inpatients of ADOJS at Department of Integrative Medical Rheumatism, General Hospital of Chengdu Military Region, of which 28 patients had knee joint effusion. Besides, 30 healthy volunteers who received physical examination at our hospital were recruited as the normal control group, and 30 patients with osteoarthritis (OA) who had knee joint effusion were recruited as the synovial fluid control group. The expression levels of IL-17 in serum and the synovial fluid were detected by enzyme linked immunosorbent assay (ELISA), and contents of ESR and CRP were detected in RA patients. Then correlation analyses were performed between levels of IL-17 and contents of ESR and CRP.

RESULTSCompared with the normal serum control group, the expression levels of IL-17 in serum of RA patients significantly increased (P < 0.05). Compared with the serum of RA patients and the synovial fluid of OA patients, the expression levels of IL-17 in the synovial fluid of RA patients significantly increased (P < 0.05). The expression levels of IL-17 in serum of RA patients were not correlated with ESR or CRP (r = 0.092, -0.082; P > 0.05), and the expressional levels of IL-17 in the synovial fluid of RA patients were not correlated with ESR or CRP (r = 0.113, -0.034; P > 0.05).

CONCLUSIONSIL-17 was the main effector cytokine of Th17 cells. The expressional levels of IL-17 significantly increased in serum and the synovial fluid of RA patients of ADOJS, but with no correlation to ESR or CRP. It indicated that IL-17 participated in the occurrence and development of RA. Concrete mechanisms needed to be further proved in larger samples.

Aged ; Arthritis, Rheumatoid ; blood ; diagnosis ; metabolism ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Case-Control Studies ; Female ; Humans ; Interleukin-17 ; blood ; metabolism ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Synovial Fluid ; metabolism