BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.

Objective To investigate the influence of different timing of surgery on patients with poor-grade intracranial aneurysm. Methods The clinical data of 86 patients with aneurysmal subarachnoid hemorrhage (Hunt-Hess gradeⅣ toⅤ)were analyzed retrospectively. The patients were divided into an ultra-early treatment group (the operative time within 24 h after onset;n=40),an early treatment group, (24 to 72 h after onset;n=27),and middle and late treatment group (>72 h after onset;n=19)according to the different timing of surgery. The prognosis of patients at 6 months after procedure was evaluated with the modified Rankin Scale (mRS)scores. The differences of the rate of good prognosis (mRS 0 to 2)and mortality in patients of the 3 groups were compared. Results (1 )The ratios of good prognosis in the ultra-early treatment group,early treatment group,and middle and late treatment group were 55. 0%(n=22),33. 3%(n=9),and 21. 1%(n=4),respectively. There were significant differences (P<0. 05). The mortality rate of the ultra-early treatment group (7. 5%)was lower than that of the early treatment group (25. 9%) or middle and late treatment group (42. 1%). There were significant differences (P<0. 05). (2)There were significant differences in the rate of good prognosis in different timing of surgery in patients with gradeⅣ(P<0. 05);there was no significant difference in the rate of good prognosis in patients with grade Ⅴ(P>0. 05). The mortality ratios in patients of the 3 groups were 2/18,4/11,and 6/11,respectively. There were significant differences (P<0. 05). Conclusion Ultra-early operation may benefit part of the patients with poor-grade aneurysm,particularly in patients with gradeⅣ. As for the patients with grade Ⅴ, ultra-early operation may help to reduce short-term mortality rate,however,the rate of vegetative state is high.

Objective Severe bronchial stricture due to endobronchial tuberculosis is often accompanied by complex complication,such as obstructiv pneumonia,destroyed lung and bronchiectasis.Its treatment is very diffucult.The present report is to investigate and analyze the indication and efficacy of surgical treatment of bronchial stricture due to severe endobronchial tuberculosis.Methods Reviewed the clinico-pathological records documenting the surgical outcomes in 81 bronchial stricture due to severe endobronchial tuberculosis who underwent lobectomy or pneumonectomy enrolled in our hospital between January 1990 and December 2010.There were 29 male and 52 female.Mean age was(36 ± 12) years (ranged 16-66 years).The three most common reasons of surgery were bronchial stricture accompanied by pulmonary atelectasis,destroyed lung and bronchiectasis(76 cases,93.8％).79 cases had elective operation,whereas one patients required emergency surgery.Pueumonectomy in 51,lobectomy in 16,sleeve resection in 11,segmental resection in 2,and exploratory thoracotomy in 1.If frozen pathological examination showed that endobronchial tuberculosis remained in the bronchial stump,it was covered with muscle flaps,including intercostal muscle flap in 6 cases,latissimus dorsi muscle flap in 5 cases,serratus anterior muscle flap in 5 cases.The mean operative time was 3.2 h (range between 2 h and 5.5 h) and the blood loss averaged 546 ml (range between 100ml and 4 000 ml).The post operative hospital stay averaged(12 ±8)days.Results No intraoperative or early postoperative death occurred.Nine patients developed complications,including BPF in 2,pulmonary infection in 2,empyema in 1,hemorragic shock in 1,hemothorax in 1,incision infection in 1,chylothorax in 1.All 9 cases recovered well after treatment.Pathological examination showed that tuberculosis bronchial remained in the brinchial stump in 13 cases.Neither BPF nor empyema occurred in all the 13 cases.Multivariate analysis revealed that destroyed lung was significant risk factor of postoperative complication.There were 3 late deaths.Five year survival rate was 96.2％.Conclusion Surgical treatment is still the recommeded treatment modatity for bronchial stricture caused by endobronchial tuberculosis due to its excellent results.It should be performed in time when the drug and intraluninal treatment were no effect for avoiding of being progeressed into destroyed lung.

Objective To determine the prognosis and staging non small cell lung cancer (NSCLC) that extends across the fissure into adjacent lobe after surgery.Methods 3752 patients with histopathologically confirmed non small cell lung cancer (NSCLC) received surgical reeessetion from January,1997 to April,2007.Among them,163 patients have a tumor invasion beyond fissure.After matching by pathologic TNM staging (7th),326 patients whose tumor defined in a single lobe were eligible for analysis.Results Histopatholngic staging of matched patients was I a:10 patiens(6.1％ ),I b:79 patients (48.5％),Ⅱa:5 patients (3.1％ ),111:44 patients (27.0％) and Ⅲa:25 patients( 15.3％ ).5 years survival in patients with stage 1 tumors crossing the interlobar fissure was 51％,while in patients not cross the interlobar fissure was 63％ ( P ＜0.05 ).There was no difference in survival for tumors stage Ⅱa and above with regard to importance of interlobar extension.The T2 tumor extending across a lung fissure had a reduction in survival compared with T2 tumor not cross the lung fissure and similar to the T3 tumor without the fissure invasion.Conclusion Our results suggest that TNM staging should be modified for tumor extends the fissure into adjacent lobe.

Objective To compare the clinic characteristics, recurrences and prognosis in patients with stage Ⅰ bronchioloalveolar carcinoma (BAC) and adenocarcinoma of the lung. Methods The data of 56 patients with stage Ⅰ BAC and 169 patients with stage Ⅰ adenocarcinoma were analyzed retrospectively. Results The overall 1-, 3-, 5- year survival rates were 94.7%, 83.5% and 61.2%, respectively. Compared with adenocarcinoma of the lung, BAC showed a better survival rate(x2 =6.36, P =0.012). After surgery patients with BAC were prone to develop intrathoracic recurrence, and adenocarcinoma was equal between intrathoracic recurrence and extrathoracic metastasis. The rate of intrathoracic recurrence and extrathoracic metastasis between BAC and adenocarcinoma was significantly different (14/16 vs. 27/59, x2 =8.85, P=0.004). In both group, preoperative asymptomatic patients had better survival rate(x2 = 7.28, P = 0.007; x2 = 6.07, P = 0. 014). Univariate analysis indicated that sex, age(< 60 years and ≥60 years), location of tumor and smoking history did not significantly influence survival in patients with stage Ⅰ BAC or adenocarcinoma (P > 0.05). Conclusion The prognosis of stage Ⅰ BAC is superior to that of stage Ⅰ adenocarcinoma. BAC is prone to develop intrathoracic recurrence, and adenocarcinoma is equal between intrathoracic recurrence and extrathoracic metastasis. Early diagnosis of lung cancer could improve long-term survival.

This research established an HPLC method for determination of six C-Glycoside flavones of warer-soluble total flavonoids from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, and studied the antitumor activity of the warer-soluble total flavonoids. The HPLC system consisted of Kromasil 100-5 C18 (4.6 mm x 250 mm, 5 microm) column and a solution system of methanol, acetonitrile and 0.5% formic acid gradient elution at a flow rate of 0. 8 mL x min(-1) and the wavelength of detector was at 334 nm. The column temperature was 25 degrees C. The antitumor activity of water-soluble flavonoids was assayed using HepG2 cell as the tested cell. The linear ranges of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabinosylapigenin were 0.25-2.53, 0.12-1.20, 0.37-3.69, 0.16-1.63, 0.19-1.92, 0.14-1.42 microg, respectively. The average recoveries (n = 6) were 99.6% (RSD 0.87%), 100.2% (RSD 2.0%), 99.6% (RSD 1.8%), 97.9% (RSD 1.5%), 98.8% (RSD 1.2%), 98.6% (RSD 1.2%), respectively. After exposure in 24, 48, 72 h, the total flavonoids showed inhibitory effect on the proliferation of HepG2 cells with IC50 as the evaluation index, the IC50 values of 1.89, 1.71, 1.51 g x L(-1), respectively. The method is quick, simple and accurate with good re- producibility, and can be used for determination of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabino- sylapigenin in the warer-soluble total flavonoids from L lophanthoides var. gerardianus. The warer-soluble total flavonoids from L lophanthoides have inhibitory effect on the proliferation of HepG2 cells.
Antineoplastic Agents, Phytogenic ; analysis ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Flavones ; analysis ; pharmacology ; Humans ; Isodon ; chemistry ; Monosaccharides ; analysis ; pharmacology

OBJECTIVETo observe the difference in the efficacy on the symptoms of bronchial asthma at the chronic persistent stage between acupoint heat-sensitive moxibustion and western medicine with Seretide.

METHODSSixty-four cases were randomly divided into a heat-sensitive moxibustion group (32 cases) and a western medication group (32 cases). In the heat-sensitive moxibustion group, the sensitized points located between Feishu (BL 13) and Geshu (BL 17) or in the region 6-cun lateral from the 1st and the 2nd intercostal spaces of the chest were selected. The heat-sensitive moxibustion was adopted, continuously for 8 days, once per day. In the later 22 days of the 1st month, 12 treatments should be ensured. Two months later, 15 treatments should be guaranteed each month. The time of each treatment was 30 to 90 min. Totally 50 treatments were required. In the western medication group, Seretide inhaler was adopted, one inhalation each time, twice per day, for 3 months totally. The asthmatic symptoms were scored for the patients in two groups and the comparison was made between the two groups.

RESULTSAfter 3 months of treatment, the asthmatic symptom scores were all improved for the patients in the heat-sensitive moxibustion group and the western medication group as compared with those before treatment (both P < 0.05). In 6 months of follow-up visit, the asthmatic symptom scores in the heat-sensitive moxibustion group were stable, but those in the western medication group were reduced, there was significant difference between the two groups (P < 0.05).

CONCLUSIONThe acupoint heat-sensitive moxibustion effectively relieves the clinical symptoms for the patients with bronchial asthma at the chronic persistent stage. Its efficacy is similar to that of Seretide inhaler. But the long-term efficacy of the heat-sensitive moxibustion is much better.

Acupuncture Points ; Adolescent ; Adult ; Albuterol ; analogs & derivatives ; therapeutic use ; Androstadienes ; therapeutic use ; Asthma ; therapy ; Chronic Disease ; Drug Combinations ; Female ; Fluticasone-Salmeterol Drug Combination ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Moxibustion ; methods

OBJECTIVESTo examine the anti-oncogenic effects of promyelocytic leukemia (PML) on bladder cancer and to explore its molecular mechanisms of growth suppression.

METHODSWild-type PML was transfected into bladder cancer cells (5637 cell) and expressed in a replication-deficient adenovirus-mediated gene delivery system and introduced into human bladder cancer cells (5637 cell) in vitro and in vivo. The effect and mechanisms of the PML gene in cell growth, clonogenicity, and tumorigenicity of bladder cancer cells were studied using in vitro and in vivo growth assays, soft agar colony-forming assay, cell cycle analysis, apoptosis assay and in vivo tumorigenicity assay.

RESULTSOverexpression of PML in 5637 cells significantly reduced their growth rate and clonogenicity on soft agar. PML suppressed bladder cancer cell growth by inducing G1 cell cycle arrest and apoptosis. Adenovirus-mediated PML (Ad-PML) significantly suppressed the tumorigenicity and growth of bladder cancer cells. Intratumoral injection of Ad-PML into tumors induced by 5637 cells dramatically suppressed their growth.

CONCLUSIONSThe results indicated that overexpression of PML protein may promote efficient growth inhibition of human bladder cancer cells by inducing G1 cell cycle arrest and apoptosis, and adenovirus-mediated PML (Ad-PML) expression efficiently suppresses human bladder cancer growth.

Adenoviridae ; Animals ; Apoptosis ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Humans ; Male ; Mice ; Mice, Nude ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; Promyelocytic Leukemia Protein ; Transcription Factors ; analysis ; Transfection ; Tumor Cells, Cultured ; Tumor Suppressor Proteins ; Urinary Bladder Neoplasms ; pathology

OBJECTIVETo clone and express VP1-VP4 genes encoding the structural proteins of Coxsackie virus A16 and analyze the antigenicity of the expressed recombinant proteins.

METHODSThe VP1-VP4 cDNAs were amplified with RT-PCR from the extracted viral RNA and cloned into pMD19-T vectors. The VP1-VP4 genes were inserted to the multi-cloning sites of the plasmid pQE30a, and the protein expressions in E. coli M15 were induced by IPTG. After purification by washing with 8 mol/L urea under denaturing condition, the recombinant proteins were identified by Western blotting and ELISA for their immunogenicity against rabbit antisera of Coxsackie virus A16 and enterovirus 71, respectively.

RESULTSThe recombinant VP1-VP4 proteins were highly expressed in E. coli M15. The purified proteins could be recognized by rabbit antiserum of Coxsackie virus A16 and showed cross reactivity with the rabbit antiserum of Enterovirus 71.

CONCLUSIONThe recombinant Coxsackie virus A16 VP1-VP4 proteins obtained possess good antigenicity.

Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; classification ; genetics ; immunology ; Cloning, Molecular ; Enterovirus A, Human ; genetics ; Gene Expression ; Genetic Vectors ; Plasmids ; Recombinant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the expression and significance of Cyclin D1 in oral squamous cell ma (OSCC).

METHODSA immunohistochemistry method, Envosion, was employed to test the manifesting Cyclin D1 in pathological slices of 50 OSCC cases and 10 normal cases, and the results was treated with statistical lysis.

RESULTSIn 50 OSCC cases, Cyclin D1 mainly manifested in karyon, and a little in cytoplasm. manifesting rates of Cyclin D1 in the samples was 80.0%, which was significantly higher than the manifesting of 20.0% in normal oral mucous membrane (P < 0.01). The manifestation of Cyclin D1 was correlated with rent pathological grades, clinical phases and lymph node metastasis (P < 0.05).

CONCLUSIONThe abnormal tation of Cyclin D1 is closely related with the occurrence and development of OSCC. Therefore, it can subsidiary index for OSCC treatment and prognosis.

Carcinoma, Squamous Cell ; Cyclin D1 ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Mucosa ; Mouth Neoplasms ; Prognosis