Objective To investigate the effects of pentoxifylline on the cell proliferation of, collagen synthesis and TGF-β1 expression by human fibroblasts derived from keloid. Methods Skin samples were obtained from the lesions of 3 patients with keloid and normal skin of 3 human controls followed by primary culture of fibroblasts. Fibroblasts of 5th to 8th generation were cultured with pentoxifylline of 0.1 to 3 g/L for various durations. Then, MTT assay was performed to detect the cell proliferation of fibroblasts, double antibody sandwich-enzyme linked immunosorbent assay (ELISA) to measure the expression of TGF-β1, and reversetranscription PCR to examine the mRNA expressions of procollagen Ⅰ and Ⅲ in these fibroblasts. Results The pentoxifylline of 0.1 to 2 g/L markedly inhibited the proliferation of fibroblasts derived from keloid lesions and normal skin, in a dose- and time-dependent manner, with the strongest effect observed in fibroblasts treated with pentoxifylline of 2 g/L. A significant reduction was induced in the TGF-β1 mRNA expression in keloidand normal skin-derived fibroblasts by pentoxifylline of 0.5 to 2 g/L (all P < 0.01), and in the mRNA expression of procollagen Ⅰ and Ⅲ by pentoxifylline of 1 and 2 g/L (P < 0.05 or 0.01). Concretely, the relative mRNA expression level of procollagen Ⅰ and Ⅲwas 0.873 ± 0.077, 0.571 ± 0.050 respectively in keloid fibroblasts respectively, and 0.473 ± 0.035, 0.370 ± 0.045 in the control fibroblasts, after treated with pentoxifylline of 1 g/L, 0.750 ± 0.036 and 0.433 ± 0.045 respectively in keloid-derived fibroblasts, 0.390 ± 0.030 and 0.250 ±0.123 respectively in the control fibroblasts, after treated with pentoxifylline of 2 g/L, significantly lower than that in the keloid-derived (1.216 ± 0.061 and 0.953 ± 0.060) and control (0.836 ± 0.080 and 0.776 ± 0.041) fibroblasts without treatment. Conclusion Pentoxifylline shows an evident suppressive effect on the cell proliferation of, as well as the expression of TGF-β1 and procollagen Ⅰ and Ⅲ in fibroblasts derived from keloid lesions and normal skin.

BACKGROUND:In recent years,pentoxifyIline has been found to have a wide range of anti-fibrosis capacity However,there are few studies explore the suppress effect of pentoxifyIline on fibroblasts in human keloid.and the maximum inhibitory concentration remains poorly understood.OBJECTIVE:To study the effect of pentoxifyIline on proliferation activity of human keloid fibroblasts and to select the maximum inhibitory concentrationMETHODS:Human keloid fibroblasts were used as original cells,passaged till the 5~(th) to the 8~(th) generations.and then divided into the experimental and control groups.PentoxifyIline with concentrations of 0.1,0.25,0.5,1.0,2.0 and 3.0 g/L were added to the experimental group.The effects of different concentrations of pentoxifylline on proliferation of keloid fibroblasts were detected by MTT chromometry.RESULT AND CONCLUSION:Compared with the control group,the inhibitory effect of pentoxifylline on the proliferation of keloid fibroblasts was more evident in the experimental group(P＜0.05)The inhibition rates of pentoxifylline on proliferation of keloid fibroblasts showed apparently time-and dose-effect relationships within the concentration of 0.1-2.0 g/L.which presented a greatest level at 96 hours after culture.The maximum inhibitory rate was 53 37%,and the concentration was 2.0 g/L in the experimental group.Consequently,pentoxifyIline plays a notable inhibitory role in the proliferation of human keloid fibroblasts with concentration of 2.0 g/L at 96 hours after culture.

Objective To reveal inpatient service utilization and medical burden in disease-caused poor rural families,for policy references.Methods By means of convenience sampling method,face-toface interview was made to inpatients from such families in Miyi County of Sichuan Province,Binhai County of Jiangsu Province and Xingtai County of Hebei Province.Results Totally 453 effective questionnaires were collected,with an effective rate of 96.7％.Respondents feature low income and fund raising capability,while 75.3％ of them pay their inpatient costs out-of-pocket,17.7％ pay with loans from others and 7.1％ pay by financial assistance from relatives;but they tend to go to higher level hospitals and have higher financial burdens for diseases.Conclusions Rural hospitalization resources should be optimized and integrated,levels of the New Rural Cooperative Medical System be improved,the catastrophic health care system be perfected and the health insurance coverage be gradually expanded for the low-income and marginalized population.

Objective To incestigate the fetures of cranial CT and MRI in the patients with eclamptic encephalopathy.Methods The CT and MRI findings of eight cases of eclamptic encephalopathy with the charge of CT,MRI appearance of FLAIR(fluid attenvated inversion-recovery),DWI(difussion weighted imaging),ADC(apparent diffusion coefficient)were analyzed retrospectively.Results Of eight patients,5 cases had abnormal finding in the cranial CT with showed symmetric plaque-like low-attenuated lesions in cortex and subcortical white matter of parietal and occipital lobes in six cases;eight cases had abnormal findings in the cranial MRI,the lesions were demonstrated as slightly hypointensity on T1WI and slightly hyperintensity on T2WI and remarkably hyperintensity on FLAIR,and iso or slightly hyperintensity on DWI,and remarkably hyperintensity on ADC.The lilateral parietal occipital lobes and cerebellar hemisphere and Brain Stem were the more common sites.Conclusions The only characteristric findings of eclamptic encephalopathy in MRI and CT imaging studies is vasogenic edema and reversible,especially in the subcortical white matter of the parietal and occipital lobes bilaterally,and cereballar hemisphere et al;especially cranial CT showed symmetric plaque like low-attenuated lesions of posterior brain;FLAIR,DWI and ADC of MRI can be helpful for early diagnosis and diffenential diagnosis,prognosis and curative effect of hypertensive encephalopathy.

Objective To evaluate the value of cystography with low-concentration contrast medium in diagnosing small tumor of urinary bladder.Methods Cystography in 187 cases with bladder disease from 1992 to 2001 was performed using 6%～8% Meglumine Diatrizoate 100～120 ml under TV monitored.Radiogrames of A-P position and bilateral oblique position were taken when the focus was found.The radiogram on patient’s position at head low was adopted when necessary.Results Of 187 cases,106 cases of bladder tumors were detected,including 8 cases of small bladder tumor(≤1.0 cm in diameter),3 cases were misdiagnosed,the detectable rate was 73.0%.All cases were confirmed by operation and pathologiy.In these 8 cases,1 case was adenocarcinoma,5 cases were transitional epithelia cell carcinoma,2 cases were papilloma.The X-ray appearances were nipple-like or cauliflower shape with filling defect,a narrow pedicel could be occasionally found in papilloma or transitional epithelia cell carcinoma and it had somewhat movement when changed patient’s position.Conclusion Cystography with low-concentration contrast medium is a non-injury procedure in diagnosis of small tumor of urinary bladder.

Objective To evaluate the effect of resveratrol on the growth of an established A431 xenogratt tumor in nude mice.Methods The model of human skin squamous cell carcinoma was established by inoculating A431 cells in log-phase growth into the left axillary fossa of Balb/c (nu/nu) nude mice.After 7-8 days,60 mice bearing human A431 skin squamous cell carcinoma xenografts were randomly and equally divided into 6 groups:blank control group receiving no treatment,negative control group treated with intraperitoneal sodium chloride physiological solution,positive control group treated with intraperitoneal cyclophosphamide,high-,medium-and low-dose resveratrol groups treated with intraperitoneal resveratrol of 40,20 and 10 μg per gram body weight per day,respectively.Tumor size was measured at a 4-day interval during the treatment course.After 14-day treatment,the mice were sacrificed.Xenograft tumors were removed from these mice and subjected to weight measurement,pathological examination by hematoxylin and eosin (HE) staining and apoptosis detection by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Western blot was conducted to quantify the protein expression of apoptosis-related factors,including phosphorylated extracellular signal-regulated protein kinase (p-ERK),p53 and caspase 3.Data were processed by SPSS 13.0 software,and statistical analysis was carried out by analysis of variance and Pearson correlation analysis.Results By the end of treatment,the xenograft tumor volume was (1153.56 ± 255.41) mm3,(1001.69 ± 115.08) mm3,(1206.80 ± 175.88) mm3,(1342.28 ± 211.12) mm3,(1642.34 ± 225.85) mm3 and (1564.32 ± 156.49) mm3,and the weight was (1.84 ±0.30) g,(1.72 ± 0.39) g,(1.96 ± 0.40) g,(2.67 ± 0.73) g,(3.16 ± 0.52) g,and (3.33 ± 0.59) g,respectively in the positive control group,high-,medium-and low-dose resveratrol group,negative control group and blank control group.Significant differences were observed in the xenograft tumor volume (F =16.00,P ＜ 0.05) and weight (F =19.15,P ＜ 0.05) among the 6 groups.According to the tumor weight,the growth of tumor was inhibited by 45.57％,37.97％ and 15.51％ respectively in the high-,medium-and low-dose resveratrol groups.Increased apoptotic index was observed in the positive control group,high-,medium-and low-dose resveratrol groups compared with the negative control group and blank control group (36.79 ± 8.86,33.15 ± 6.00,18.09 ±3.92 and 10.53 ± 4.20 vs.3.87 ± 1.63 and 2.73 ± 1.61,F =93.26,P ＜ 0.05).Analysis of variance showed that the protein expressions of p-ERK,p53 and caspase 3 were all higher in the three resveratrol groups than in the negative control group and blank control group (F =6.65,6.78,11.56,respectively,all P ＜ 0.05).The protein expression of p53 was statistically correlated with p-ERK (r =0.68,P ＜ 0.05) and caspase 3 (r =0.56,P ＜0.05).Conclusions Resveratrol shows an inhibitory effect on the growth of human A431 skin squamous cell carcinoma xenografts in nude mice,likely by increasing p53 expression and inducing tumor cell apoptosis via the activation of MAPK/ERK pathway.

With the increasing number of cancer patients, the aging population, the shortage of medical resources and other problems in China, the demand for palliative care is gradually increasing. However, nursing staff are the main implementors of palliative care, and their cognitive level of palliative care is closely related to the quality of palliative care service and the outcome of patients. By summarizing assessment tools of palliative care knowledge, attitudes and skills, this review aims to provide a reference for developing appropriate palliative care assessment tools for nursing staff in China.

ssociated genes,especially down-regulated expression of T cell mediated function genes,in patients with PE indicates that the etiology of PE might be related to viral infection.

Objective: To establish human cancer cell strains with stable Cas9 expression, and to validate the gene editing activity of Cas9 for simple gene editing in future study. Methods: Fifteen cancer cell lines of different tissue origins were infected with pLv-EF1α-Cas9-Flag-Neo or pLv-EF1α-Cas9-Flag-Puro by lentivirus and clone selection was employed to screen Cas9 stably expressed cancer cell lines. Afterward designed guide RNA vectors targeting TSC22 gene were transiently transfected into 3 of cell lines, and subsequently the gene editing activity of Cas9 was evaluated by genomic PCR, sequencing and Western blot. Results: Sixty-nine human cancer cell strains with stable Cas9 expression from different cancers were established, and by transient transfection with designed guide RNA, long fragment deletion was detected in TSC22 gene. Conclusions Sixty-nine human cancer cell strains are successfully established with stable expression of Cas9 protein and gene editing activity. These cell strains may be employed in large-scale drug screening, screening of new drug targets and gene function investigation.

Objective: To construct the third generation chimeric antigen receptor based on a novel humanized anti-HER2 H1-2 scFv, and to investigate the specific cytotoxicity of H1-2 CAR modified T lymphocytes(CAR-T) against HER2⁺ tumor cells. Method: The expression cassette of the third generation CAR gene and anti-HER2 H1-2 scFv were constructed and cloned into lentivirus transfer plasmid, and then the third generation H1-2 CAR was transduced into human T lymphocytes using lentivirus.Enzyme linked immunosorbent assay was used to detect the expression of cytokines IL2, and LDH release assay was used to detect the cytotoxic effect of the H1-2 CAR-T.Finally, NOD/SCID mice and HER2⁺ breast cancer cell line SKBR3 were used to detect the anti-tumor effect of H1-2 CAR-T in vivo. Results: The third generation H1-2 CAR was successfully constructed.H1-2 CAR-T secreted high dose of IL2 after confrontation with HER2⁺ breast cancer cells.In vitro, the cytolytic rate of H1-2 CAR-T on high expression HER2⁺ tumor cells was significantly higher than that in low expression HER2 or non-expression HER2 tumor cells. At the efficacy to target ratio of 20, the cytolytic rate of H1-2 CAR-T against breast cancer cell SK-BR-3 could reach (90.1±2.8)%, while the cytolytic rate of H1-2 CAR-T against HER2^- breast cancer cell MDA-MB-231 was only (13.5±4.7)%. In the mouse xenograft tumor model, H1-2 CAR-T cells inhibited breast cancer growth in vivo.At the end of the experiments, the average tumor weight in the H1-2 CAR-T cell treatment group was (0.7±0.1) g, the non-transfected T cell therapeutic group was (1.2±0.2) g, and the PBS group was (1.2±0.2) g. There was significant difference between the H1-2 CAR-T therapeutic group and the non-transfected T cell therapeutic group (P<0.05). However, there was no significant difference between the non-transfected T cell therapeutic group and the PBS treatment group (P>0.05). Conclusion The HER2-sepcific H1-2 CAR-T cells specifically kill HER2 positive cells, and further studies on CAR-T cells for the treatment of HER2⁺ cancers are useful.