A white-rot fungi Rigidoporus sp.W-1 which could produce laccase was isolated. The fermentation conditions in shaking flasks were investigated. The optimal carbon source was wheat bran and (NH 4) 2SO 4 was the optimal nitrogen source. The components of the medium were optimized by orthogonal experiment. When W-1 was cultured under the optimum conditions, the activity of laccase could get to 7.1U/mL in 7 days.A great amount of crude laccase could be obtained by adding fresh medium to the 7 days old mycelium.

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

OBJECTIVETo determine the minimum number of lymph nodes that should be examined for the UICC/AJCC TNM classification of gastric cancer.

METHODSThe clinical and pathological data of four hundred and thirty-six patients underwent curative resection for gastric cancer were analyzed by Chi-square and Student-Newman-Keuls test.

RESULTSThe pN0 patients with 1 to 4, 5 to 9 examined nodes showed significantly lower survival rate than those with 10 to 14, 15 or more examined nodes (P < 0.05), and the patients with 10 to 14 examined nodes had as good a prognosis as those with 15 or more examined nodes. In the pN1, pN2 categories, the patients with 1 to 4, 5 to 9 and 10 to 14 examined nodes tended toward significantly lower survival rates than those with 15 or more examined nodes (P < 0.05). Among the patients who were classified as stage II, the survival rate of those with 10 to 19 examined nodes was significantly lower than that with 20 or more examined nodes. Among the patients classified as stage III, those with 5 to 9, 10 to 19 and 20 to 29 examined nodes had significantly lower survival rates than those with 30 or more examined nodes (P < 0.05).

CONCLUSIONSThe number of lymph nodes examined has significant prognostic impact within each pN category of gastric carcinoma. The minimum number of lymph nodes to examine in order to determine pN0 classification can be reduced from 15 to 10. For tumor with lymph node metastasis, the minimum number should be 15. In stage II, 20 or more nodes should be examined, and in stage III and IV 30 or more.

Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Prognosis ; Stomach Neoplasms ; classification ; mortality ; pathology ; Survival Rate

This study was aimed to prepare a reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human B to O blood group conversion. Based on the construction of recombinant E. Coli (DE3) which can express α-galactosidase, the inducing time and inducer concentration were optimized for high expression of α-galactosidase. Then, the expression products in supernatant were purified by cation and anion exchange column chromatography. The purified α-galactosidase was used to treat B group red blood cells in phosphate buffer (pH 6.8) for 2 hours to prepare O group red blood cells. The results showed that the optimal inducing conditions for α-galactosidase expression were IPTG 0.1 mmol/L, 37°C and 2 hours. The specific enzyme activity of purified protein increased from 0.42 U/mg to 2.1 U/mg as compared with pre-purification. And, the conditions of B to O blood group conversion were 26°C, pH 6.8 (neutral pH condition) and 2 hours. Moreover, 225 µg of the enzyme could converse 1 ml B red blood cells to O completely. It is concluded that the technology of expression and purification of recombinant α-galactosidase has been established, and the purified protein can converse B red blood cells to O completely, which means that an effective enzyme conversing B red blood cells to O has been obtained.
ABO Blood-Group System ; immunology ; Bacteroides fragilis ; enzymology ; Cloning, Molecular ; Escherichia coli ; metabolism ; Humans ; Recombinant Proteins ; biosynthesis ; alpha-Galactosidase ; biosynthesis

OBJECTIVETo explore an exercise method for the prevention and treatment of the patients with shoulder and back fasciitis.

METHODSFrom 2006.8 to 2008.3, 120 patients with shoulder and back fasciitis were randomly divided into control group (n = 60, including 21 females and 39 males, the average age was (47.0 +/- 12.0) years, and the average course of disease was (14.1 +/- 12.0) months) and treatment group (n = 60,including 19 females and 41 males, the average age was (43.7 +/- 9.9) years, and the average course of disease was (16.4 +/- 13.4) months). The patients in the control group received massage therapy and the ones in the treatment group were treated with massage therapy and horizontal bar exercise. After 3 weeks treatment, the curative effects of the patients in two groups were observed. All the patients were followed up for 6 to 26 months, the recurrence were observed.

RESULTSAfter 3 weeks treatment, the scores of pain, sense of heaviness, strip sign, tenderness, shoulder and back function of the patients in two groups had significant differences compared with those before treatment (all P < 0.01). After treatment, the scores of pain, sense of heaviness, strip sign, tenderness, shoulder and back function of the patients in the treatment group were lower than those in the control group (P < 0.05). After 6 to 26 months following-up, the rate of recurrence in the treatment group was lower than that in the control group (P < 0.01).

CONCLUSIONHorizontal bar exercise is a simple, no expense and effective method in the prevention and treatment of shoulder and back fasciitis, which can improve the effect of the treatment and reduce the rate of recurrence.

Adult ; Back Pain ; etiology ; prevention & control ; therapy ; Exercise Therapy ; methods ; Fasciitis ; prevention & control ; therapy ; Female ; Humans ; Male ; Middle Aged ; Shoulder Pain ; etiology ; prevention & control ; therapy ; Treatment Outcome

OBJECTIVETo study therapeutic effects of embracing knee and rolling lumbar (EKRL) training on the pain and the function of lumbar of patients with degenerative osteoarthritis of lumbar spine.

METHODSFrom 2006.8 to 2008.1, 150 patients with degenerative osteoarthritis of lumbar spine were randomly divided into the control group, EKRL training group and the combined group. In the control group (n=50), there were 16 females and 34 males, the average age was (58.8 +/- 10.2) years and the average course of diseases was (16.3 +/- 14.9) months. In the EKRL training group (n=50), there were 13 females and 37 males, the average age was (60.1 +/- 8.2) years and the average course of disease was (17.1 +/- 15.3) months. In the combined group (n=50), there were 15 females and 35 males, the average ages was (59.9 +/- 11.4) years and the average course of diseases was (19.8 +/- 17.2) months. The patients in three groups were treated with Meloxicam tablets, EKRL training and two above methods combination respectively. After 3 weeks treatment, the curative effect, the visual analogous scores and the ODI indexes were observed. All the patients were followed up and the duration ranged from 6 to 24 months; the changes of average ODI indexes were observed monthly.

RESULTSAfter 3 weeks treatment, the therapeutic effects in three groups had obvious difference after Ridit analyses: there were significant differences between the combined group and other two groups (P<0.01), but no obvious differences existed in other two groups. Compared with before treatment, the visual analogue scale scores and the ODI indexes in three groups reduced significantly (P<0.01). After treatment, the visual analogue scale scores and the ODI indexes of the combined group had significant differences compared with those of other two groups (P<0.01), but the ODI indexes of other two groups had no obvious differences. After 6 to 24 months following-up, the monthly changes of average ODI indexes of EKRL training and combined groups had obvious differences compared with that of the control group (P<0.01).

CONCLUSIONEKRL training is a simple, no expense and effective methods for preventing and treating degenerative osteoarthritis of lumbar spine.

Adult ; Aged ; Case-Control Studies ; Exercise Therapy ; methods ; Female ; Follow-Up Studies ; Humans ; Knee ; Lumbar Vertebrae ; Male ; Middle Aged ; Osteoarthritis ; prevention & control ; therapy

OBJECTIVETo investigate the clinical presentations and pathologic features of undifferentiated sarcoma of the prostate with cartilage metaplasia, and to clarify its category.

METHODSWe analyzed the clinical data of a case of undifferentiated sarcoma of the prostate with cartilage metaplasia treated by surgical resection. The tumor tissue was subjected to routine HE and immunohistochemical staining, its histological structure and immunohistochemical expression were observed under the light microscope, and relevant literature on its manifestations was reviewed.

RESULTSThe case was pathologically diagnosed as gray prostate tumor, with chondrosarcomatous and undifferentiated malignant mesenchymal components under the light microscope. Immunohistochemical staining revealed vimentin (+), local CD117 (+/-), SMA (-), Des (-), myoglobin (-), CD34 (-), CK7 (-), and CK8 (-). Tumor metastasis was found 2 months after the operation, and the patient died 4 months later.

CONCLUSIONUndifferentiated sarcoma of the prostate with cartilage metaplasia is a very rare and highly malignant aggressive tumor, which can be diagnosed by biopsy and immunohistochemistry.

Adult ; Cartilage ; pathology ; Humans ; Male ; Metaplasia ; Prostate ; pathology ; Prostatic Neoplasms ; diagnosis ; pathology ; Sarcoma ; diagnosis ; pathology

αGal, a xenotransplantations antigen (XTA), can lead to hyper acute reaction (HAR) in xenotransplantation. α-Galactosidase from B. fragilis is a novel galactosidase belong to CAZy GH110 which can clear the terminal αGal from branched and linear oligosaccharides. This study was purposed to investigate the removal effect of a novel α-galactosidase on α-Gal XTA on surface of red blood cells. The αGal XTA from the red blood cells of cattle, pig, dog and rabbit was digested by using recombinant α-galactosidase; the α-Gal antigens on surface of cells was detected by flow cytometry. The results showed that the XTA was disappeared completely or mainly. It is concluded that the novel α-galactosidase is a potential enzyme to remove the XTA on the surface of xenotransplants and can be used to overcome the HAR in xenotransplantation.
Animals ; Antigens, Heterophile ; immunology ; Cattle ; Dogs ; Epitopes ; Erythrocytes ; immunology ; Macaca mulatta ; Mice ; Mice, Inbred BALB C ; Rabbits ; Swine ; Transplantation, Heterologous ; alpha-Galactosidase ; immunology

BACKGROUND: Percutaneous vertebroplasty (PVP) has been proved to be effective for osteoporotic vertebral compression fractures (OVCF); however, bone cement leakage-related complications occur frequently. OBJECTIVE: To systematically evaluate the safety and complications of PVP with high-viscosity and low-viscosity bone cement in the treatment of OVCF. METHODS: We performed a systematic search by computer in PubMed, EMBASE, Cochrane Library (December, 2016) and CNKI, CBM, VIP and WanFang databases from January, 2000 to December, 2016; meanwhile, the relevant Chinese and English orthopedic journals were retrieved manually. Randomized controlled trials (RCTs) concerning the use of high-viscosity and low-viscosity bone cement in the PVP for OVCF were collected and systematically reviewed using the method recommended by the Cochrane Collaboration. The Meta-analysis was performed by using the RevMan5.2. RESULTS AND CONCLUSION: Six relevant RCTs were included, involving 554 patients (high-viscosity bone cement was used in 278 cases and low-viscosity bone cement in 276 cases). Meta-analysis results showed that postoperative pain symptoms were alleviated in all the 554 cases, and there was no significant difference in the amount of adjacent vertebral fractures between two approaches (P > 0.05). However, there was a significant difference in the visual analog scale score, Oswestry disability index value, Cobb angle, and bone cement leakage rate between high-viscosity and low-viscosity cement groups (P < 0.05). To conclude, PVP with either high-viscosity or low-viscosity bone cement can relieve pain rapidly in OVCF patients, and both approaches are safe and effective. However, high-viscosity bone cement has a lower cement leakage rate and better outcomes in visual analog scale score, Oswestry disability index, Cobb angle and cement leakage prevention as compared with low-viscosity bone cement.

BACKGROUNDHuman group O red blood cells have great benefit in specialized transfusion areas such as armed conflict and natural calamity. The group B antigen differs structurally from group O antigen only by the addition of one terminal alpha-linked galactose residue. In this study we aimed to remove the terminal galactose from group B red blood cell to get group O red blood cell.

METHODSalpha-galactosidase cDNA was cloned by RT-PCR from Catimor coffee beans grown on Hainan Island of China. The vector for alpha-galactosidase cDNA expression was constructed and transferred into Pichia pastoris cells by electroporation. The transgenic cells were cloned by fermentation and the recombinant alpha-galactosidase was purified by ion exchange chromatography. After studying the biochemical characters of alpha-galactosidase, we have used it in converting human erythrocytes from group B to group O.

RESULTSThe purity of recombinant alpha-galactosidase was higher than 96%, which was thought to be suitable for the use of blood conversion. Enzymatically converted human group O red blood cells (ECHORBC) exhibited membrane integrity, metabolic integrity, normal cell deformation and morphology. There were no coagulation between ECHORBC and any group of human blood. The ECHORBC will keep normal structure and function for a period of 21 days at 4 degrees C in monoammoniumphosphate nutrient solution. Experiments with Rhesus monkeys and gibbons showed that transfusion of enzymatically converted erythrocytes was safe.

CONCLUSIONECHORBC can be easily obtained from group B red blood cell by alpha-galactosidase digestion. This study suggests that ECHORBC could be transfused to patients safely and efficiently.

ABO Blood-Group System ; classification ; metabolism ; Animals ; Blood Transfusion ; Cloning, Molecular ; Coffee ; enzymology ; Erythrocytes ; metabolism ; Humans ; Macaca mulatta ; Quality Control ; Recombinant Proteins ; isolation & purification ; pharmacology ; alpha-Galactosidase ; immunology ; isolation & purification ; pharmacology ; toxicity