Purpose:To evaluate the feasibility of irradiat ion to the metastatic retroperitoneal lymph nodes using three-dimensional confo rmal radiotherapy (3-DCRT) and to assess the irradiation side effects. Methods:From Jun. 1997 to Sep. 2000, 52 patients of metastatic retroperitoneal lymph nodes were treated with 3-DCRT.These patients had receive d radical therapy for their primary tumors. 50 of them had been proved by pathol ogical examination. Radiotherapy was designed by Elekta company’s 3-D treatmen t plan. Dose volume histography (DVH) was used to verify and optimize the radiat ion plan. Using 5～6 conformal portals,the total irradiation dose was 40 Gy an d delivered at 4～5Gy per fraction every other day,to a total of 8～10 fractions in 16～20 days. The immediate responses and side effects were evaluated accordi ng to the WHO and RTOG criterion system. Results:According to Karnofsky criterion system the median scor e was raised 21.3. Three months after irradiation, the complete ,partial and tot al response rates were 38.6%(27/70), 54.3%(38/70)and 92.9%(65/70),respect ively. The early response was related to the target volume and pathological clas sification. The smaller the target volume, the better the response(? 2 = 1 5.211,P=0.004).The anaplastic carcinoma cases had better results than the squamous carcinomas and adenocarcinomas(? 2 =13.455,P=0.009).The 1- ,2- and 3- year survival rates were 59.6%(31/52)、23.1%(12/52)and 9.6%(5/ 52),respectively. Conclusions:Three dimensional conformal radiotherapy can be con sidered as an effective and feasible approach for the treatment of metastatic r etroperitoneal lymph nodes,and no serious complications were observed.

Objective To study the appearance of magnetic resonance imaging (MRI) in early stages of cartilage degeneration and to detect its values.Methods Intra articular injection of 5 units of papain causing a reversible loss of cartilage proteoglycan in the New Zealand rabbit knees.Rabbits were scanned with magnetic resonance imaging,using a 0 3 T Hitachi magnet with 16 cm coil.Gradient echo sequences and spin echo sequences were performed in the sagittal planes at 0,24,48 and 72 hours after intra injection of papain.Then signal intensity and thickness of cartilage were measured.The proteoglycan content was measured biochemically and histochemically.Results The cartilage thickness and signal intensity decreased significantly in treated knees compared with control knees at 24 and 48 hours ( P 0 05) after injection of papain.These changes on magenetic resonance (MR) images were consistent with the changes of proteoglycan concentration measured by biochemical analysis and histochemical staining of cartilage.Conclusion It is possible for MRI to detect the early stages of cartilage degeneration.

Aim To investigate the effects of leptin on ?1(Ⅰ) collagen mRNA expression and protein production, and the roles of Janus kinase/signal transducers and activators transcription(JAK/STAT) signaling transduction pathway in increased ?1(Ⅰ) collagen mRNA expression stimulated by leptin in activated hepatic stellate cells(HSCs).Methods Firstly, ?1(Ⅰ) collagen mRNA expression and protein production as well as JAK1 and STAT3 phosphorylation induced by leptin at different doses in a human HSC cell line, LX-2 were determined by RT-PCR, ELISA, and Western-blot.Secondly, the effects of JAK1 inhibitor AG490 on JAK1 phosphorylation and ?1(Ⅰ) collagen mRNA expression stimulated by leptin were detected by Western blot and RT-PCR. Thirdly, the roles of AG490 and transfection with STAT3 antisense oligonucleotide(STAT3-ASON) in STAT3 phosphorylation after leptin were detected by Western blot. Finally, the effect of transfection with STAT3-ASON on ?1(Ⅰ) collagen mRNA expression after leptin was measured by RT-PCR.Results Leptin increased ?1(Ⅰ) collagen mRNA expression and protein production in a dose-dependent manner in LX-2, reaching a maximal level at 80 ?g?L-1 leptin. In addition, phosphorylation of JAK1 and STAT3 after leptin exhibited a time-dependent effect. Besides, JAK1 inhibitor AG490 completely blocked JAK1 and STAT3 phosphorylation and increased in ?1(Ⅰ) collagen gene expression after leptin in LX-2. Transfection with STAT3-ASON blocked STAT3 phosphorylation and increased ?1(Ⅰ) collagen mRNA by leptin in LX-2.Conclusion Leptin had a direct action on liver fibrogenesis by stimulating ?1(Ⅰ) collagen mRNA expression and protein production in activated HSC, and JAK/STAT signaling transduction pathway was involved in the process. JAK1 inhibitor AG490 and transfection with STAT3-ASON blocked the transduction pathway effectively in LX-2. Leptin may be an important factor in the development of hepatic fibrosis. Activated JAK1 and STAT3 signaling in human HSC line provided a novel molecular target for therapeutic intervention of liver fibrosis.

Objective To study the correlation of fingerprint chromatograms of Cinnamomi Ramulus formula granules, decoction pieces and water decoction by HPLC; To investigate the difference of main chemical constituents among different forms. Methods The Diamonsil C18 column (4.6 mm × 250 mm, 5 μm) was used with mobile phase of acetonitrile and 0.1% phosphoric acid at the flow rate of 1.0 mL/min, with detection wave of 280 nm and temperature of 30 ℃. The detection of 10 batches of Cinnamomi Ramulus formula granules, 10 batches of decoction pieces and 10 batches of water decoction were established respectively. Results Totally 12 peaks in the HPLC fingerprint chromatogram from 10 batches of formula granules could be tracked in the water decoction; 10 peaks in the HPLC fingerprint chromatogram could be tracked in the decoction pieces. Three components, such as protocatechuic acid, coumarin and cinnamic acid were verified. Conclusion The main chemical components of Cinnamomi Ramulus formula granules and water decoction are basically the same, and the common component contents have similar proportion.

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Acupuncture Therapy ; methods ; Adult ; Aged ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Tennis Elbow ; therapy

Objective: To observe the therapeutic efficacy of astragalus polysaccharide (APS) in combination with three-dimensional conformal radiotherapy (3D-CRT) for elderly patients with non-small cell lung cancer (NSCLC). Methods: A total of 80 elderly patients with I～IV stage NSCLC were randomly divided into two groups. The 40 patients in the therapeutic group received radiotherapy in combination with APS. The 40 patients in the control group received radiotherapy alone. Both groups received 3D-CRT with a total dose of 50～ 70Gy, 2.0Gy/fraction, once a day, 5 times per week. The patients in the therapeutic group were treated with radiotherapy combined with injection of APS (250mg in 5% glucose) or normal saline (500 mL) intravenously once a day until the end of radiotherapy. The short-term efficacy and patients' quality of life were evaluated. The T-lymphocyte subpopulation and peripheral blood leukocyte count were also measured after treatment. Results: The short-term effective rates of the therapeutic group and the control group were 87.5% and 72.5%, respectively, without significant difference between the two groups (P＞0.05). The decrease of peripheral blood leukocyte count after treatment in the control group was significant (P＜0.05). In the therapeutic group, the T-lymphocyte subsets CD8 and CD4/CD8 were improved after treatment, with a significant difference (P＜ 0.05). But in the control group, no significant changes in T-lymphocyte subsets CD8 and CD4/CD8 were observed after treatment (P＞0.05). Patients' quality of life in the therapeutic group was superior to that in the control group, with a significant difference (P＜0.05). Conclusion: APS in combination with 3D-CRT can reduce the side effects of radiation and improve the quality of life of elderly patients with NSCLC.

BACKGROUND:Under the in vitro conditions of cell harvesting, culture, and transplantation, whether bone marrow stromal cells (BMSCs) can be effectively applied in local gene therapy remains unclear.OBJECTIVE: To construct a recombinant eukaryotic expression plasmid carrying human bone morphogenetic protein-7 (hBMP-7) gene, and to expect to enhance osteoinductive properties of rabbit BMSCs transfected.DESIGN, TIME AND SETTING: A cell-genomics in vitro observation was performed at the Laboratory of Scientific Research, Second Affiliated Hospital of Harbin Medical University between July 2006 and July 2007.MATERIALS: Human healthy fresh placental tissue was provided by the Department of Gynaecology and Obstetrics, Second Affiliated Hospital of Harbin Medical University. Written informed consent was obtained from the women. One healthy male New Zealand rabbit was provided by the Laboratory Animal Center, Harbin Medical University.METHODS: hBMP-7 gene was cloned from human placental tissue to construct a recombinant eukaryotic expression plasmid carrying hBMP-7 gene by conjugating with eukaryotic expression vector pcDNA3.1. BMSCs were isolated from rabbit bone marrow and cultured in vitro. Then they were divided into 3 groups: pcDNA3.1-hBMP-7-transfected, pcDNA3.1 -transfected, and untransfected. 5×106 BMSCs were inoculated into a 60 mm3 flask containing antibiotic-free medium 1 day prior to transfection.MAIN OUTCOME MEASURES: RT-PCR and immunohistochemistry were employed to detect hBMP-7 expression in BMSCs, alkaline phosphatase activity, hydroxypreline content, and osteocalcin production in each group. RESULTS: After 72-hour transfection, a 1.3 kb fragment was seen in the pcDNA3.1-hBMP-7-transfected group, showing brown granules in the endochylema, but not seen in the pcDNA3.14ransfected and untransfected groups. ALP activity in the pcDNA3.1-hBMP-7-transfected group significantly increased at 2 days after transfection, peeked at 8 days, and still increased at 10 days. At each time point, alkaline phosphatase activity, hydroxyproline content, and osteocalcin production were significantly higher in the pcDNA3.1-hBMP-7-transfected group than in the pcDNA3.1 -transfected and untransfected groups (P<0.05 or P<0.01).CONCLUSION: Recombinant eukaryotic expression vector pcDNA3.1- BMP-7 was constructed successfully. Results indicated that hBMP-7 was expressed in BMSCs sufficiently and was involved in inducing differentiation of BMSCs into osteoblasts. The method would provide substantial basement for hBMP-7 gene therapy.

ObjectiveTo evaluate the diagnostic value of ultrasound or CT guided percutaneous Trucut needle biopsy on the diagnosis of pancreatic tumors.Methods One hundred and twenty-four patients clinically diagnosed as pancreatic cancer without pathological diagnosis underwent percutaneous pancreatic biopsy by using Trucut needle under ultrasound or CT guidance.ResultsOne hundred and nine procedures of ultrasound-guided biopsy and 15 procedures of CT-guided biopsy were performed,and one patient received 2.3times of punctures.Tissue samples were obtained in all 124 patients,the diagnostic accuracy was 95.2％,among them 115 were adenocarcinoma,5 were cystadenoma,2 were metastasis cancer,1 was cancer of unknown origin and 1 was normal.The sensitivity,specificity,and accuracy were 99.2％ 100％,and 99.2％,respectively.Transient serum amylase increase was observed in 3 patients; 5 patients' abdominal pain aggravated,but all recovered with conservative management.One patient was found to have tumor seeding on the spot of insertion after 34 days.No other major complications occurred.ConclusionsUltrasound or CTguided percutaneous pancreatic 16 ～ 18G Trucut needle biopsy is a safe and simple procedure with excellent diagnostic value for pancreatic cancer.