[Objective]To compare the purity and extraction rate of four purecollagen type I products prepared from various biomaterials by a limited enzyme digestion method for the use in tissue engineering.[Method]Bovine cortical bone, bovine achilles tendon, porcine achilles tendon and porcine skin were splitted into pieces of 0.2-0.5 mm. After being immersed in glacial acetic acid, they were extracted with pepsin. Then the crude products were dissolved, centrifuged, dialyzed and freeze drying to prepare pure collagen type I. The final products were confirmed by absorbance, amino acid analysis and'SDS-PAGE electrophoresis comparing them with the products of Sigma Company.[Result]The wave length of maximum absorbance of the final products was 230 nm, and the amino acid analysis and SDS-PAGE electrophoresis confirmed that the final products were collagen type I. The purity of product extracted from bovine cortical bone was the highest (96.12%) and higher than that from Sigma Company. The extraction rate of bovine achilles tendon collagen was the highest (75.34%).[Conclusion]Collagen type I of higher purity and higher extraction rate can be prepared using a limited enzyme digestion method.And the product from bovine cortical bone is better than the others,which has a promising prospect.

The aim of this paper is to apply Raman spectroscopy technique to develop rapid quantitative models for five kinds of Traditional Chinese Medicine containing CaCO3. In the experiment, Raman spectras of 67 batch of sample including Otolithum Sciaenae, Galaxeae Os, Ophicalcitum, Calcite, Stalactite and their mixture which had different content of CaCO3 were collected, and the quantitative models were established by using an improved siPLS to optimize the characteristic spectral bands and using the CaCO3 contents which were measured by EDTA titration method as references. Compared with the results by EDTA titration, the established quantitative model for CaCO, content showed a prediction result that the average relative deviation of the prediction results is 2. 71% and the average recovery rate was 100.46%, when the content is between 0.465 4-0.999 7, and when the characteristic spectral bands of 1 290-1 280, 730-714, 700-690, 660-650, 465-460, 455-445, 405-385 cm(-1) had been optimized. The result also showed that the model using Raman spectroscopy and based on an improved siPLS can get a rapid determination for contents of 5 kinds of Traditional Chinese Medicine containing CaCO3.
Calcium Carbonate ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Models, Statistical ; Plants, Medicinal ; chemistry ; Spectrum Analysis, Raman ; methods

The influenza A virus matrix protein2 gene(M2)which deleted transmembrane region was amplified by overlap extending PCR,and the multiepitope gene of hemagglutinin(HA)was PCR amplified with seven continuous synthesized segments by designing primer.The two gene segments were separately cloned into pMD18T vector to sequence analysis and prokarytic expression vector pET28a+ to construct the recombinant plasmid pET28a+M2dHA.The recombinant plasmid was transformed into E.coli BL21(DE3),and the high expression strain was obtained by screening monoclones.The recombinant protein existed as inclusion bodies,which accounted about 45% of the total cellular protein.The inclusion bodies were washed with 1% Triton X100 solution twice,and dissolved in 8 mol/L urea solution.The solution protein was purified by Ni+2 affinity chromatography,and refolded by dilution renaturation,then purified by Q Sepharose FF cation exchange column.The purity of the protein was over 90% by HPLC analysis.The result of Western blot showed it has good antigenicity and specificity.These results strongly supported for the further study of the broadspectrum influenza virus subunit vaccine.

OBJECTIVETo retrospectively investigate the clinical characteristics of sternal insufficiency fractures (SIFs) of post-menopausal women.

METHODSFindings on the clinical presentation, associated diseases, and imaging of SIFs in 17 postmenopausal women admitted to our hospital between February 1999 and January 2009 were reported.

RESULTSTwelve patients complained of severe pain in their anterior chest. Other symptoms included cough (5 cases), dyspnoea (3 cases), breathlessness (3 cases), and wheeze (2 cases). Four patients had no discomfort. The sternums of 11 cases were tender to palpation. Seventeen patients had osteoporosis. Other associated diseases were chronic obstructive pulmonary disease (7 cases), rheumatoid arthritis (3 cases), systemic lupus erythematosus (1 case), asthma (1 case), and thoracic vertebral fracture (13 cases). Nine patients had received glucocorticoid treatment. The fractures were located in the body of the sternum in 15 patients, in the manubrium in 1 patient, and in the manubriosternal junction in 1 patient. Displaced fracture was present in 13 cases. Lateral radiography of the sternum showed a fracture line in 14 patients. In the remaining 3 cases, other imaging examinations such as bone scan, computed tomography or magnetic resonance imaging demonstrated the presence of a fracture.

CONCLUSIONSOsteoporosis, glucocorticoid therapy, chronic obstructive pulmonary disease, and rheumatoid arthritis might be risk factors for SIFs. SIFs should be considered in the differential diagnosis of chest pain.

Aged ; Aged, 80 and over ; Arthritis, Rheumatoid ; complications ; epidemiology ; Cohort Studies ; Female ; Fractures, Bone ; diagnosis ; epidemiology ; etiology ; Fractures, Stress ; diagnosis ; epidemiology ; etiology ; Glucocorticoids ; adverse effects ; therapeutic use ; Humans ; Middle Aged ; Osteoporosis, Postmenopausal ; chemically induced ; complications ; epidemiology ; Postmenopause ; physiology ; Pulmonary Disease, Chronic Obstructive ; complications ; epidemiology ; Retrospective Studies ; Risk Factors ; Sternum ; injuries ; pathology

OBJECTIVETo observe mid- and long-term changes in the histopathology and electron microscopic characteristics of the acellular dermal matrix engrafted with thin split-thickness skin autograft.

METHODSTwenty-three biopsy samples were collected from 17 patients undergoing extremity scar resection, who received subsequent grafting using allogenic dermal matrix dressed with thin split-thickness skin autografts. Six months to 2 years after the grafting, the grafts were sampled for histopathological and electron microscopic observations of the layer of the epidermis, thickness of the basal membrane, structural components of the dermis, and infiltration of fibroblasts and revascularization. The data were compared with those of the normal skin samples from the patients.

RESULTSOnly the number of epidermal layers showed statistically significant difference between the skin grafts and the normal skin (16.33-/+5.89 vs 26.57-/+3.46, P=0.007). The thickness of the basal membrane of the skin grafts was similar to that of normal skin, and no significant difference was found in the number of fibroblasts and newly generated capillaries between them.

CONCLUSIONThe mid- and long-term histopathology and ultrastructures of the composite skin graft in the extremities are similar to those of normal skin, suggesting satisfactory effect of the skin grafts.

Adolescent ; Burns ; surgery ; Child ; Child, Preschool ; Cicatrix ; surgery ; Dermis ; transplantation ; ultrastructure ; Female ; Follow-Up Studies ; Graft Survival ; Humans ; Male ; Skin ; ultrastructure ; Skin Transplantation ; methods ; Skin, Artificial ; Transplantation, Autologous

Breast cancer has the highest incidence rate among all women's malignant tumors worldwide.Surgery,radiotherapy and chemotherapy are three major treatments,while most patients showed adverse effects or complications during or after the treatment,including lymphedema,gastrointestinal reactions and leukopenia,which cause severe impact on patients' recovery and quality of life.Moxibustion has been used and certified to alleviate adverse effects of surgery or chemoradiotherapy for breast cancer.We have summarized literatures in recent years and suggest more systematic research in the future for the underlying mechanism.

OBJECTIVETo investigate and analyze the variation trends in the pathological composition of thyroid cancer patients treated in Tianjin Cancer Hospital from 1954 to 2009.

METHODSTo retrospectively analyze the incidence and clinical features of different pathological types of thyroid cancers in 4342 patients between different time periods from 1954 to 2009.

RESULTSIn the four main pathological types of thyroid cancers, the component ratio of papillary thyroid cancer in every period was 68.1%, 78.3%, 81.3%, 82.1%, 85.8%, respectively, while the morbidity of patients with papillary thyroid carcinoma concurrent with Hashimoto's thyroiditis was increased, so was the proportion of tumors in diameter < or = 2 cm. The proportion of follicular thyroid carcinoma and anaplastic thyroid carcinoma was decreasing accordingly; however, the proportion of medullary thyroid carcinoma did not change significantly.

CONCLUSIONSThe pathological classification of the thyroid carcinoma patients has significant changes in the 4342 cases treated in our Hospital from 1954 to 2009. The proportion of papillary carcinoma is increased, while that of follicular carcinoma and anaplastic carcinoma is decreased. The reasons might attribute to the improved level of consultations and iodized diet or other factors.

Adenocarcinoma, Follicular ; epidemiology ; pathology ; Carcinoma ; epidemiology ; pathology ; Carcinoma, Medullary ; epidemiology ; pathology ; Carcinoma, Papillary ; epidemiology ; pathology ; China ; epidemiology ; Female ; Hashimoto Disease ; complications ; epidemiology ; pathology ; Humans ; Incidence ; Male ; Retrospective Studies ; Thyroid Neoplasms ; complications ; epidemiology ; pathology ; Tumor Burden

For determination the ionic mechanisms of the hypoxic acclimatization at the level of channels, male Spradue-Dawley rats were divided into two groups: control normoxic group and chronic intermittent hypoxic group [O2 concentration: (10 +/-0.5)%, hypoxia 8 h a day]. Using whole cell patch-clamp technique, voltage-gated potassium channel currents (IK(V)) were recorded in freshly isolated pulmonary arterial smooth muscle cells (PASMCs) of rat with acute isolated method. The effect of acute hypoxia on IK(V) of PASMCs from chronic intermittent hypoxia group was investigated to offer some basic data for clarifying the ionic mechanisms of the hypoxic acclimatization. The results showed: (1) In control normoxic group, after acute hypoxia free-Ca(2+) solution, the resting membrane potential (Em) of PASMCs was depolarized significantly from -47.2+/-2.6 mV to -26.7+/-1.2 mV, and the IK(V) of PASMCs was decreased significantly from 153.4+/-9.5 pA/pF to 70.1+/-0.6 pA/pF, the peak current percent inhibition was up to (57.6+/-3.3)% at +60 mV, and current-voltage relationship curve shifted to the right. (2) In chronic intermittent hypoxic group, the IK(V) of PASMCs was decreased significantly by exposure to intermittent hypoxia in a time-dependent manner, appeared to start on day 10 and continued to day 30 (the longest time tested) of hypoxia, and current-voltage relationship curve shifted to the right in a time-dependent manner. (3) Compared with the control normoxic group, the percent IK(V) inhibition by acute hypoxia was significantly attenuated in the chronic intermittent hypoxia group and this inhibition effect declined with time exposure to hypoxia. The results suggest that K(V) inhibition was significantly attenuated by chronic intermittent hypoxia, and this effect may be a critical mechanism of the body hypoxic acclimatization.
Animals ; Cell Separation ; Hypoxia ; complications ; physiopathology ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Potassium Channels, Voltage-Gated ; antagonists & inhibitors ; Pulmonary Artery ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

Objective:To observe the clinical effect of acupuncture for persistent allergic rhinitis (PAR).Methods:A total of 154 PAR patients were randomized into a treatment group and a control group,78 cases in the treatment group received acupuncture at meridian points,while 76 cases in the control group received acupuncture at non-meridian points.The treatment was done every other day,3 times a week for a total 4 weeks.The total nasal symptom score (TNSS),the total non-nasal symptom score (TNNSS) and the rhinoconjunctivitis quality of life questionnaire (RQLQ) were measured before the treatment and after 2 weeks and 4 weeks of treatment,as well as 1 month and 3 months after the treatment to compare the clinical effect between the two groups.Results:After 4 weeks of treatment,the total effective rate in the treatment group was 92.3％,versus 76.3％ in the control group,showing a statistically significant difference (P＜0.05).Intra-group comparisons of TNSS,TNNSS and RQLQ scores after 4 weeks of treatment showed statistically significant differences (all P＜0.05).Between-group comparisons of TNSS,TNNSS and RQLQ score after 4 weeks of treatment and in follow-up visits showed statistically significant differences (all P＜0.05).Conclusion:Acupuncture is effective for PAR,and acupuncture at meridian points can produce a better effect than acupuncture at non-meridian points.

Objective To investigate the effects of different dialysates on expression of protein kinase C-? (PKC?) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKC? specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKC? mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group, normal control group and fluid A+powder B+rottlerin group (P0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKC?. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.