OBJECTIVE:To optimize ultrasonic extraction and purification technology of solanesol from tobacco leaf. METH-ODS:Using extraction rate and transport rate of solanesol as indexes,single factor test was used to investigate liquid-solid ratio, ultrasonic extraction temperature and time,ultrasonic power and extraction times,and the amount of soap alkali lye(volume ratio of soap alkali lye to extraction liquid),acidizing fluids (volume ratio of acidizing fluids to soap alkali lye extract),extraction times of purification technology. Optimized technology was validated,and the purity of solanesol was calculated;the amount of ex-tracted solanesol was compared between this method and traditional extraction method (spending 30 h),solvent continuous cyclic extraction (spending 5-6 h). RESULTS:Optimized extraction technology was as follows as volume ratio of soap alkali lye to ex-traction liquid 1∶14,ultrasonic extraction temperature 70 ℃,ultrasonic extraction time 60 min,ultrasonic power 120 W,extract-ing for 3 times;optimized purification technology was as follows as volume ratio of soap alkali lye to extraction liquid 2∶35,vol-ume ratio of acidizing fluids to soap alkali lye extract 2∶14,extracting for 4 times. In validation test,extraction rate,transport rate and purity were 92.45%(RSD=0.46%,n=3),79.88%(RSD=0.30%,n=3)and 55.86%(RSD=0.40%,n=3). The amount of solanesol extracted with 3 methods were 52.22,45.22 and 26.10 mg/g. CONCLUSIONS:The optimized technology is simple and stable,costs less time and saves source with high extraction amount and purity,which is suitable for production,extraction and purification of solanesol from tobacco leaf.

Objective To evaluate the feasibility of the DNA microarray method used in detecting the drug resistance of Myco-bacterium tuberculosis by comparing the traditional proportion method and the DNA microarray method for detecting the drug re-sistance of Mycobacterium tuberculosis.Methods 54 strains of Mycobacterium tuberculosis isolated from clinical specimens in our hospital from January 2012 to March 2013 were randomly extracted and their resistance to isonicotinic acid hydrazide (INH)and rifampicin(RFP)was detected by the DNA microarray method and the proportion method.The detection results were performed the comparative analysis.Results With the proportion method as the golden standard,the coincidence rates of the DNA microarray method for detecting the Mycobacterium tuberculosis resistance to INH and RFP were 75% and 91.0% respectively.Conclusion The DNA microarray technique is suitable for the rapid screening of clinical first-line drug resistant Mycobacterium tuberculosis.

Objective To investigate the expression of muhidrug resistance-related protein 1 (MRP1),lung resistance-related protein(LfuP)and glutathione S-transferase-π(GST-π)in carcinoma of the gallbladder and cholangiocarcinoma. Methoils MRP1,LRP,GST-πwere measured in experimental group(18 cases of carcinoma of the gallbladder,36 CSSeS Of cholangiocarcinoma)and control group(13cases of cholecystitis and cholangeitis)by immunohistochemistry.Statistical analysis used chi-square test and spearman test. Results The positive rate of MRP1,LRP,GST-π in carcinoma of the gallbladder and eholangiocarcinoma were 72%(13/18),78%(14/18),61%(11/18)and 86%(31/36),75%(27/36),69%(25/36),respectively,significantly higher than those of 23%(3/13),23%(3/13),23%(3/13)(X2=4.5,P＜0.05)in control group.The expression of LRP[93%(13/14)]in pafients＞60 years old was significantly higher than 64%(14/22)in patients younger than 60 yrs old(x2=3.9,P ＜0.05).In addition,their expression was not related to gender,age,staging,tumor differentiation and lymph node metastasis(P＞0.05).The expression of MRP1 was related with tllose Of GST-π,Spearman correlation coefficient=0.569(P＜0.05).Conclusions MRP1,LRP,GsT-π were over expressed in various degrees in carcinoma Of the gallbladder and cholangiocarcinoma witllout chemotherapy.and related to the primary muhidrug resistance Of cholangiocarcinoma and carcinoma of the gallbladder.

Objective To investigate the relationship between various clinicopatholngic factors and prognosis of early gastric cancer. Methods The clinical data of 459 patients with early gastric cancer who had undergone gastrectomy combined with extended (D2) lymphadenectomy at Zhongshan Hospital from January 2002 to October 2007 were retrospectively analyzed. Survival was calculated using the Kaplan-Meier method, and clinicopathologic factors such as age, sex, tumor size, gross morphology, tumor differentiation, depth of invasion, lymphatic vessels involvement and lymph node metastasis were analyzed using the Cox regression model. Univariate analysis was done by Log-rank test. Results Univariate analysis demonstrated that tumor size and differentiation, depth of invasion, lymphatic vessels involvement and lymph node metastasis significantly affected survival in patients with early gastric cancer (χ~2 = 8.476, 6.210, 4.014, 14. 197, 55.027, P < 0.05). The status of lymph node metastasis was an independent predictor of survival in patients with early gastric cancer, and the more metastatic lymph nodes detected, the greater the influence on the prognosis. Conclusions Lymph node metastasis is the most important factor influencing the prognosis of early gastric cancer. Appropriate lymph node dissection is necessary, especially for patients with risk factors associated with lymph node metastasis.

BACKGROUND:Diffusion conditions of bone cement in vertebral fracture line may be one of the main factors affecting the therapeutic effect of percutaneous vertebroplasty, but there are less related studies. OBJECTIVE:To study the effect of diffusion conditions of bone cement in vertebral fracture line on the therapeutic outcomes of percutaneous vertebroplasty. METHODS: CR and MRI data of 77 patients with T1-L2 osteoporotic vertebral compression fractures, 28 males and 49 females, aged 55-86 years, undergoing percutaneous vertebroplasty were analyzed. Al the patients were divided into test group (n=53, bone cement diffused wel in the vertebral fracture line) and control group (n=24, bone cement dispersion was unsatisfactory). Visual analogue scale, Oswestry disability index and Cobb angle change in the two groups were measured and compared before and after operation. RESULTS AND CONCLUSION:There was no difference in the visual analogue scale score, Oswestry disability index and Cobb angle between the two groups before operation, but these parameters were al improved significantly in the two groups after 2 days and 6 months of operation (P< 0.05). The visual analogue scale score and Oswestry disability index were significantly lower in the test group than the control group at 2 days after operation (P < 0.05), but there was no difference between the two groups at 6 months after operation. The Cobb angle and vertebral colapse rate became lower in the test group than the control group at 6 months after operation (P < 0.05), but there was no difference in the re-fracture rate between the two groups. These findings indicate that poor bone cement dispersion in the fracture line can affect the relief of short-term pain and dysfunction and it can increase the possibility of long-term secondary vertebral colapse.

BACKGROUND: In repair of nerve defect with allogenic nerve graft, to reduce immune rejection is one of the key problems. At present, the main approach is to reduce antigenicity of grafted nerve segment and apply generally immune inhibitor.OBJECTIVE: To observe the effects of freeze/thaw treatment and local application of transforming growth factor beta 1 (TGF-β1) plasmid on frozen nerve allograft.DESIGN: Randomized controlled animal experiment was designed.SETTING: Department of Hand Surgery, Union Hospital, Tongji Medical College Affiliated to Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Tongji Medical College of Huazhong University of Science and Technology from January 2003 to December 2004, in which 40 Wistar healthy and adult rats were employed,from different delivery and were randomized into experimental group and control, 20 rats in each one.METHODS: Transforming growth factor-β1 (TGF-β1) plasmid and frozen allogenic sciatic nerve were prepared. In experimental group and control,sciatic nerve was cut off 2.0 cm in length, in the foramen 0.5 cm beneath piriformis. The nerve defect was repaired with pre-frozen allogenic nerve 2.0 cm in length. In experimental group, TGF-β1 plasmid was injected in local muscle and two broken ends of nerve. In the control group, physiological saline of equal volume was injected. In the 6th and 12th weeks, the samples were collected from 10 rats in each group for sectioning, staining,axonal counting and statistical analysis.RESULTS: No any animal was died in experiment and all of animals entered result analysis. In the 6th weeks, in the control group, mild edema appeared among axons on the grafted segment of nerve and in the experimental group, there was no edema among axons and the regenerated nerve numbers were close to the normal. In the 12th week, in the experimental group, the entire grafted nerve segment was basically filled up by the regenerated axons;myelinated nerve fiber was arranged in order and both axons and myelins were developed well. The regenerated axonal count in experimental group was more significantly than the control, indicating extremely significant difference [(98.6±4.8), (75.8±5.1) counts/μm2, t=2.962, P ＜ 0.01].CONCLUSION: Freeze/thaw treatment can decrease antigenicity of allogenic nerve, which provides the possibility of repair of nerve defect. Local application of TGF-β1 plasmid can provide immune inhibition locally and reduce immune rejection in the host.

BACKGROUND: Auto-neural transplantation is used widely on peripheral neurological defect, but it also has some difficulties. So some scholars try to use xenoma-neural transplantation; however, it is hard todeal with immunological rejection.OBJECTIVE: To study the effect of transforming growth factor-β1 (TGFβ1) used in local area on neural regeneration after transplantation of fresh nerve allograft.DESIGN: Randomized controlled study.SETTING: Hand Surgery Department of Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and TechnologY.MATERIALS: The experiment was conducted in the Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology between August 2001 and October 2002. Totally 60healthy adult Wistar rats from different confinements were randomly divided into three groups including experimental group, blank group and control group with 20 in each group.METHODS: TGF-β1 plasmid was prepared for using. Establishment of animal model: Sciatic nerve at the 0.5 cm deep of piriformis muscle of rats in the two groups was cut with disinfectant razor into chip regularly about 2.0 cm. The excisional nerve segment was exchanged to transplant plerosis neurological defect. TGF-31 was injected into the local muscles and bisection of nerve in the experimental group, and equal volume of saline was injected into rats in the blank group and the control group. In addition, rats in the experimental group and the blank group were not treated with any drugs, but cyclosporine A (15 mg/kg) was used to feed rats in the control group. Ten rats from each group were taken for section and staining at the 6th and the 12th week: ① Glees-luxot fast blue staining method; ② myelin sheath fast blue staining method. Axonal amount: Fields were randomly taken from the middle staining samples 12 weeks later and 1.0 mm2 interaxis-cylinder was counted under light microscope of 400 times. Comparisons among groups were analyzed with i test.MAIN OUTCOME MEASURES: Morphological observation and axonal amount of transplanted area in each group.RESULTS: Quantitative analysis of the experimental animals: Totally 60rats entered the final analysis without any loss. ① Infiltration of monocytes was observed widely in various areas of graft in the blank group;meanwhile, desiccation of myelin sheath and plenty of vacuolations were also observed, especially at the sixth week. The whole graft was infiltrated by monocyte with severe rejection. Few axis-cylinders were regenerated in the transplanted segment. At the 12th week, graft was slender, plenty of scar tissues were proliferated, edema was observed obviously, few Schwann cells and regenerated axis-cylinders were observed, and lots of regenerated axis-cylinders did not pass the whole graft. A few infiltrative monocytes were observed, and edema was observed obviously, but new vessel was formed in transplanted nerve, and regenerated axis-cylinders passed the whole graft in the experimental group and the control group.Lots of Schwann cells were observed at the 6th week; meanwhile, regenerated axis-cylinders passed the whole graft at the 12th week, a quantitative myelinization was formed, Schwann cells proliferated obviously, and edema between axis-cylinder was relieved. Numbers of peripherally regener ated axis-cylinder of nerve and remyelination in each ransplanted area were more than those in the central area, and edema between peripheral axis-cylinder was milder than that in the central area in the experimental group. ② Twelve weeks after operation, 5 rats in each group were selected to observe their fields, which were taken randomly from neural graft,under the microscope of 400 times to count 1.0 mm2 inter-axis-cylinders.Number of axis-cylinder was higher in the experimental group and the control group than that in the blank group, and the differences were significant [(78.3±4.6), (76.1±4.2) , (15.0±3.5) ,t=3.056, t=2.948, P ＜ 0.01];however, number in the experimental group was similar to that in the control group, and differences were not significant [(78.3±4.6), (76.1±4.2),t=1.982 P ＞ 0.05].CONCLUSION: TGF-β1 used in local area plays an immunosuppressive action locally, decreases host immunological rejection, increases the number of axis-cylinder, and accelerates growth of nerve.

Objective To investigate the toxicity-reducing and action-enhancing effects of total glucosides of paeony (TGP) on tripterygium glycosides (TG) in treating murine lupus nephritis (LN).Methods Chronic graft-versus-host disease (cGVHD) murine LN model was established.Modeling mice were randomly divided into three groups:LN group,TG group (in the dose of 30 mg?kg-1?d-1),and TG+TGP group (TG in the dose of 30 mg?kg-1?d-1 and TGP in the dose of 200 mg?kg-1?d-1),6 mice in each group.Another six normal mice served as control.Medication groups received corresponding medicine according to the experimental design,and mice in the normal control group and LN group received the same volume of saline,qd,for 4 weeks.After the treatment,the levels of 24-hour urinary protein excretion (UPE),blood urea nitrogen (BUN),serum creatinine (SCr),serum alanine transferase (ALT) and aspartate transferase (AST),and contents of superoxide dismustase (SOD),malondialdehyde (MDA),and glutathione peroxidase (GSH-Px) in liver tissue homogenate were measured.Meanwhile,the pathologic changes of kidneys in each group were detected.Results Compared with LN group,levels of UPE,BUN and SCr in TG group and TG+TGP group were decreased notablely (P

Protective effect of interleukin-1beta (IL-1beta) on motor neurons was studied after peripheral nerve injury. Twenty Wistar rats were divided into 2 groups randomly. The right sciatic nerve of each rat was resected. After silicon tubulization of sciatic nerve in rat, 15 microl 1 ng/ml IL-1beta and PBS solution were injected into the silicon capsule respectively. Enzyme histochemistry was performed to show acetyle cholesterase (AchE) and nitric oxide staining (NOS) activity of spinal alpha motor neurons in spinal segments 2 weeks later. Neurons were counted and the diameter and cross sectional (c/s) area of neurons were analyzed by using computer image analysis system. The results showed that as compared with the normal side, both enzyme activities significantly changed in motor neurons in PBS group. The diameter and c/s area of both neurons changed significantly too (P < 0.01). These results suggest that exogenous IL-1beta protects alpha-motor neurons from degeneration and necrosis after peripheral nerve injury.
Interleukin-1/*pharmacology ; Motor Neurons/*pathology ; Neuroprotective Agents/*pharmacology ; Random Allocation ; Rats, Wistar ; Sciatic Nerve/*injuries ; Spinal Cord/pathology

BACKGROUND:Whether there is a necessary connection or internal patterns between the amount of bone cement-recovery of vertebral height-clinical efficacy has no evidence-based medicine findings in the treatment of osteoporotic vertebral compression fractures using percutaneous vertebroplasty with high-viscosity bone cement injection.
OBJECTIVE: To observe the change of vertebral height in the elderly with osteoporotic vertebral compression fractures after percutaneous vertebroplasty with high-viscosity bone cement injection.
METHODS:A total of 110 elderly patients with osteoporotic vertebral compression fractures (139 vertebrae) were admitted at the Traditional Chinese Medical Hospital of Xinjiang Uygur Autonomous Region from January 2011 to December 2012. Al the patients received percutaneous vertebroplasty by the same group of surgeons. Bone cement at drawing stage was injected into the fractured vertebra. During the 12-month folow-up, visual analogue scale, Barthel index and vertebral height restoration were observed as evaluation indexes.
RESULTS AND CONCLUSION:The surgical treatment was done successfuly in the 110 patients (139 vertebrae). The amount of bone cement per vertebra was 3-6 mL, with a mean of 3.5 mL. At 12 months after surgery, the visual analog scale scores were decreased from 7.9 to 1.8, Barthel index was increased from 40.25 to 82.21, both of which were improved significantly (P < 0.05). After surgery, the vertebral heights at the anterior and middle parts were increased by (81.25±9.26)% and (78.22±10.65)%, respectively, and there was significant differences before ant at 24 hours, 3 months and 12 months after surgery (P < 0.05). During the folow-up, there were five vertebrae with bone cement leakage, but no clinical symptoms occurred, and no nerve injury or pulmonary embolism happened. These findings indicate that percutaneous vertebroplasty with high-viscosity bone cement injection can effectively relieve pain, restore the vertebral height, reduce the incidence of complications and shorten the recovery time in patients with osteoporotic vertebral compression fractures.