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.

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.

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.

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.

Objective To investigate the clinical and pathologic features of squamous cell carcinoma of the breast.Methods The clinical and pathologic data of 23 squamous cell carcinoma of the breast patients admitted between 1984 and 2013 to Tianjin Medical University Cancer Hospital was analyzed retrospectively.Results Primary squamous cell carcinoma of the breast was a very rare tumor accounting for 0.06％ of all breast cancers.All of the 23 patients were females aged 28 years to 87 years(median age 49 years).Average tumor size was 4.5 cm.9 patients suffered from lymph node metastasis at admission (39.1％).The positive rates of estrogen receptor (ER),progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2) were 13.6％ (3/23),4.5％ (1/23) and 0 (0/20) respectively.With a follow-up time varying from 5 months to 36 months recurrence or metastasis were found in 8 patients,and another 1 patient was found having distant metastasis at admission.Lung metastasis (7/9) was most common.6 patients died.Conclusions Squamous cell carcinoma of the breast is highly invasive,with low rate of positive receptors and early distant metastasis or recurrence after operation,and poor patients' survival.

Objective To study the clinical curative effect of Tongqiao Huoxue decoction combined with edaravone and hyperbaric oxygen for treatment of delayed encephalopathy after carbon monoxide poisoning (DEACMP). Methods A prospective study was conducted. Forty-six patients with DEACMP admitted into Yidu Central Hospital of Weifang Medical College from January 2012 to January 2014 were randomly divided into observation group (23 cases) and control group (23 cases). The basic treatments of two groups were identical. Based on the basic treatments including hyperbaric oxygen and citicoline sodium injection etc, the observation group was treated with our-self made Tongqiao Huoxue decoction which could be modified in accord to the individual differentiation of syndromes in traditional Chinese medicine (the composition of decoction included Paeoniae Radix Rubra 15 g, Chuanxiong Rhizoma 15 g, Persicae Semen 15 g, Carthami Flos 15 g, Zingiberis Rhizoma Recens 3 pieces, Jujubae Fructus 2 pieces, Moschus 0.5 g, Allium Fistulosum 1 segment). The decoction was administered orally or by nasogastric gavage, one dosage everyday for 1 month, and in the mean time, edaravone intravenously drip 30 mg was given to the observation group twice a day for 14 days. The control group was given hyperbaric oxygen and other conventional treatment for 30 days. The clinical therapeutic effect and adverse reaction were observed after treatment for 30 days. The changes of intelligent level were detected by Hasegawa dementia scale (HDS), and the changes of latency of P300 were measured by electromyologram/evoked potential instrument in two groups before and after treatment. Results The total effective rate in observation group was significantly higher than that in control group [91.3% (21/23) vs. 65.2% (15/23), P < 0.01]. Elevation of creatinine occurred in 1 case, moderate increase in alanine aminotransferase (ALT) appeared in 1 case, and both of them were reduced to normal after treatment in observation group; no adverse reaction occurred in control group. The HDS scores were significantly higher 30 days after treatment than those before treatment in the two groups [control group:13.4±2.8 vs. 6.8±2.3, observation group:20.8±3.4 vs. 6.6±2.5, both P<0.05]. The latency of P300 after treatment was significantly lower in two groups than that before treatment [control group (ms): 355.7±25.7 vs. 385.5±27.8, observation group (ms): 337.3±24.6 vs. 386.8±25.4, both P < 0.05], the change in observation group being more significant [the HDS score: 20.8±3.4 vs. 13.4±2.8, the latency of P300 (ms): 337.3±24.6 vs. 355.7±25.7, both P<0.05]. Conclusion Tongqiao Huoxue decoction combined with edaravone and hyperbaric oxygen has favorable cognitive effect on patients with DEACMP, thus, it can be used extensively in clinic.

ObjectiveTo determine the genotypes of human papillomavirus(HPV) and to estimate their correlations with recurrence of perianal and anal condyloma acuminatum(CA) among men who have sex with men(MSM) in Shanghai.MethodsTissue specimens were obtained from the lesions of perianal or anal CA in 57 MSM.Flow-through hybridization and gene chip were used to determine the genotypes of HPV in these specimens.Follow-up was carried out to observe the recurrence of CA in these patients after CO2 laser treatment.ResultsOf the 57 specimens,56(98.2％) were positive for HPV,and a total of 11 HPV subtypes were identified.The predominant genotypes were HPV 11 (42 cases,75％ ) and HPV 6 (30 cases,53.6％),sequentially followed by HPV 16 (8 cases,14.3％),HPV 18 (4 cases,7.1％),HPV 33 (2 cases,3.6％),HPV 59(2 cases,3.6％),HPV cp8304 (2 cases,3.6％),HPV 31 (1 case,1.8％) and HPV 35 (1 case,1.8％).Of the HPV-positive specimens,64.3％(36/56) harbored low risk HPV types(11 and/or 6),and 35.7 ％ (20/56) harbored high risk HPV types.Among the 20 specimens positive for high risk HPV,17(85％) harbored both low risk- and high risk-HPV genotypes.During the 6-month follow-up after CO2 laser treatment,recurrence was observed in 95％ of patients with high risk HPV and 66.7％ of patients with low risk HPV (P ＜ 0.05).ConclusionsAmong MSM with perianal and anal CA in Shanghai,the predominant genotypes of HPV are low risk types 11 and 6; high risk HPV infection is usually complicated by low risk HPV infection; the recurrence of CA is higher in patients with high risk HPV than in those with low risk HPV.

Objective To investigate the mechanism underlying the WNK4 kinasemediated inhibitory effect on BK channel. Methods Cos-7 cells were cotransfected with BK in combination with either CD4 (control group) or wild type WNK4 (WNK4-WT).Immunostaining and confocal microscopy,chemiluminescence,Western blotting analysis were then employed to determine the BK localization in cells,BK surface expression and total protein level,respectively.To further investigate whether the reduction of BK protein expression is due to an increase in degradation through a lysosomal pathway,BK protein level was determined after treated with bafilomycin A1(Baf A1),a proton pump inhibitor affecting lysosomal degradation. Results Immunostaining and confocal microscopic study showed that BK was localized both in plasma membrane and cytosol in the control group.After cells transfected with WNK4-WT,BK expression was markedly reduced.Chemiluminescent assay found that BK surface expression level was 299.9±18.6 in the control group,whereas it was significantly reduced (148.4±13.7,P＜0.01) in the WNK4-WT group.Western blotting analysis showed that total BK protein level was markedly reduced in the presence of WNK4-WT compared to the control group.WNK4-WT was shown to significantly reduce the BK total protein level (42.3％±15.2％) compared to the control group (100％) (P＜0.01).When the cells was treated with Bafilomycin A1 (Baf A1,0.5 μmol/L),WNK4-mediated reduction in BK protein was reversed (82.2％±12.1％,P＜0.05). Conclusions WNK4 inhibits total and surface protein expression of BK in Cos-7 cells whick is likely due to an increase in BK degradation through a lysosomal pathway.