This study was to investigate the inhibitory effects of single-chain variable fragment of alpha fetopro-tein (scFv-AFP)in combination with doxorubicin on the proliferation of human hepatocellular carcinoma cell lines Huh7.Huh7 cells were treated with different concentration of scFv-AFP or doxorubcin alone or their combi-nation.The inhibitory effects were detected by MTT assay,and cycle arrest and apoptosis of Huh7 cells were ana-lyzed by flow cytometry in different groups using PI and Annexin V /PI-staining respectively.Results showed that scFv-AFP,doxorubicin alone or in combinations dose-dependently inhibited the proliferation of Huh7,and a syn-ergistic effect was observed in their combined action.The combination treatment resulted in significantly higher apoptosis than those in other groups (P <0.05).scFv-AFP (40 μg/mL)markedly blocked the Huh7 cell pro-gression by arresting the cells in the G0 /G1 phase,and the percentage of cells in S phase decreased dramatically (P <0.05);and scFv-AFP combined with doxorubicin blocked the Huh7 cell progression by arresting the cells in G2 /Mphase (P <0.01).

Objective:To investigate the effect and mechanism of Wogonin on CD3AK cell proliferation and cytotoxicity to SMMC-7721.Methods:CD3AK cells were cultured from peripheral blood mononuclear cells ( PBMC) in vitro by a variety of cytokines for 7 d,and treated with different concentrations of Wogonin for 48 h.CD3AK cells proliferation was measured by CCK-8 assay.SMMC-7721 cell growth was detected by MTT.The expression of perforin (PFP),granzyme B (GrB) and CD107a on CD3AK cells were measured by flow cytometry ( FCM).The cytotoxicity to SMMC-7721 cells was detected by LDH release assay.The expression of ERK1/2 on CD3AK cells was detected by Western blot.The mobility of SMMC-7721 cells was detected with transwell chambers.The merge of SMMC-7721 cells were measured with Wound healing assay.Results:Wogonin could significantly promote CD3AK cells proliferation, especially at 3.2 mg/L (23%higher than that of control group,P<0.05).The highest cytotoxicity to SMMC-7721 was also at the con-centration 3.2 mg/L (60.4%).The expression of PFP,GrB,CD107a were significantly higher than that of control group( P<0.05).The expression of ERK1/2 was obviously improved,especially at 12.5-0.8mg/L.After treated with Wogonin 50,12.5,3.2,0.8,0.2 mg/L for 48 h,the lowest transwell cell was at 12.5 mg/L and lowest merge rate was at 3.2 mg/L.Conclusion:Wogonin could promote CD3AK cell proliferation and enhance the cytotoxicity to SMMC-7721.Wogonin could also inhibit SMMC-7721 cell growth,migration and cell merge.The mechanism may be related to activated ERK1/2 and increase the expression of PFP,GrB,CD107a.

Objective To study the effects of postnatal overfeeding and high-fat diet on blood pressure of rats,and to explore the pathophysiological mechanism underlying hypertension induced by continuous early postnatal overfeeding.Methods Male Sprague-Dawley rats were randomly divided into normal feeding group (10/litter) and overfeeding group (3/litter) on postnatal day 3 with a random number table.After weaning at postnatal week 3,the rats were randomly given standard chow or high-fat (HF) diet until week 16.Hence four groups were analyzed,namely normal feeding group,breastfed overfeeding group,post-weaning overfeeding group,and continuous overfeeding group.Body weight was continuously monitored in each week.Visceral fat pad (retroperitoneal and perigenital),systolic pressure,and heart rate were observed at week 3 and week 16.Thoracic aorta was sampled for measurement of vascular endothelial dilation function.Histological morphology was observed with HE staining,nitric oxide content of thoracic aorta was detected with nitrate reductase method.The mRNA expression of endothelial nitric oxide synthase (eNOS) in thoracic aorta was determined by real-time polymerase chain reaction.The protein expressions of eNOS and phosphorylated eNOS were determined by Western blot.Results At week 3,breastfed overfeeding rats displayed significantly larger body weight [(77.80 ± 0.57) g vs.(62.80 ±0.85) g,t =14.576,P ＜ 0.01] and visceral fat [retroperitoneal:(8.19 ± 0.49) mg/g vs.(4.92 ± 0.31) mg/g,t =5.629,P＜0.01;perigenital:(3.50 ±0.29) mg/g vs.(2.08 ±0.13) mg/g,t =4.552,P ＜0.01] compared with normal feedindg rats,and the protein expression of phosphorylated eNOS in aortic tissues was significantly reduced to week 16 (F =15.215,P ＜0.01);high-fat diet feeding after weaning further increased the body weight and fat mass in breastfed overfeeding rats.At week 16,continuous overfeeding rats showed hypertension [(149 ± 1.94) mmHg (1 mmHg =0.133 kPa),F =22.834,P ＜0.01],impaired vascular endothelial dilation function (F =7.648,P ＜ 0.05),and reduced protein expression of phosphorylated eNOS (F =15.215,P ＜ 0.01),while the post-weaning overfeeding group only had elevated blood pressure.Conclusions Overfeeding in breastfeeding period and high-fat diet after weaning leads to hypertension.The continuous decrease in phosphorylated eNOS in vascular tissues may be an important molecular process participating in the occurrence of vascular endothelial dysfunction in adults induced by postnatal overfeeding.

Objective To observe ghrelin expression in gastric tissue and serum leptin level of the early over-fed rats given ω-3 polyunsaturated fatty acids (PUFAs) diets after weaning,and explore the effects of early over-feeding and diets intervention on the metabolic syndrome in adult rats.Methods Male Sprague-Dawley rats were divided into normal feeding group (NF group,10 per litter) or over-feeding group (OF group,3 per litter) in postnatal day 3,and then different diets were given after weaning (postnatal day 21).The OF group was separately given conventional diet (OF + Con group),high-fat diet (OF + HF group),or ω-3 PUFAs (OF + ω-3 group) ; while the NF group was separated into NF + Con group and NF + HF group.Body weight and food intake were recorded every week.In week 6 and week 16,glucose tolerance test was perforfmed.Serum leptin,ghrelin,and triglyceride were assayed by enzyme-linked immuno-absorbent assay.Ghrelin mRNA and protein levels in gastric tissue were quantified by real-time PCR and immunohistochemistry.Results At week 16,energy intake,body weight and glucose intolerance in OF + HF group were significantly higher than in NF + HF group (t =-3.453,P =0.014 ; t =-6.406,P =0.000 ; F =16.249,P =0.000),and serum triglycerides,ghrelin mRNA of gastric tissue were significantly higher than those of OF + Con group (t =4.72,P =0.005 ; t =8.486,P =0.000).At week 16,the serum leptin level in OF + Con group was higher than that in NF + Con group (t =-3.274,P =0.031),also higher in OF + HF group than that in OF + Con group (t =3.028,P =0.014).There were no significant differences in serum ghrelin and the area of ghrelin immuno-positive cells in the gastric tissue among groups.The above indicators in OF + ω-3 group were not different from those of NF + Con group.Conclusions Over-feeding during the lactation period may lead to high susceptibility to obesity and disordered glucose and lipid metabolism.It can also increase serum leptin and ghrelin mRNA expression in gastric tissue,aggravate leptin resistance and feeding control disorders.Dietary ω-3 PUFAs offered protection against excessive accumulation of adipose tissue,glucose intolerance,leptin resistance,and maintained normal levels of leptin and ghrelin.

Objective To investigate the effect of neonatal overfeeding on the expression of lipid metabolic associated enzymes and molecular mechanisms in the livers of rats. Methods Male Sprague-Dawley rats were randomly assigned to litter sizes of three group ( small litters,SL group) or ten ( normal litters,NL group) on postnatal day 3. Body weight,milk intake,liver and fat pad (epididymal and retroperitonea) weight,and hepatic histological anal-ysis were recorded in week 2 and week 3,respectively. The levels of lipids were detected by the automatic biochemical analyzer. The mRNA expressions of acetyl-CoA carboxylase ( ACC) ,lipoprotein lipase ( LPL) ,liver-type fatty acid-binding protein (L-FABP), carnitine palmitoyltransferase 1(CPT1),microsomal triglyceride transfer protein (MTP), sterol regulatory element-binding protein-1c (SREBP-1c) and peroxisome proliferator activated receptorα(PPARα) in liver were determined by real time PCR;the protein expressions of SREBP-1c and PPARα were determined by Western lot. Results As early as week 2,the body weight of rats in SL group began to elevate (t=-5. 997,P<0. 001) and food intake (t=-3. 462,P=0. 002) compared with the rats in NL group,and persistent to weaning (body weight:t=-17. 019,P<0. 001;food intake:t=-2. 276,P=0. 031). By the time of 3 weeks old,SL rats increased visceral fat pad [ret-roperitonea (t=-7. 643,P<0. 001),epididymal (t=5. 997,P=0. 001)],liver weight (t=-7. 812,P<0. 001),hepatosomatic index (t=-3.829,P=0. 003) and serum triglyceride (TG) level (t=-2. 703,P=0. 022) compared with those of NL rats,as well as the level of hepatic ACC mRNA (t=-3. 751,P=0. 007),LPL (t=-2. 721,P=0. 017) and L-FABP mRNA (t=-2. 521,P=0. 026) . While CPT1 mRNA (t=-1. 531,P=0. 155) and MTP mRNA (t=-1. 741,P=0. 098) levels remained unchanged in both groups. Hepatic SREBP-1c mRNA expression increased in SL rats after 2 to 3 weeks (t=-2. 836,P=0. 016),paralleled with ACC and LPL mRNA expression;while the mRNA and protein expression of PPARα re-mained unchanged (t=-0. 854,P=0. 411). Conclusions Postnatal overfeeding can promote higher liver pad and dyslip-idemia at the time of weaning. The process may be regulated by up-regulated expression of ACC, LPL and L-FABP. SREBP-1c may be participated in the regulation of ACC,a rate-limiting enzyme involved in lipogenesis.

Objective　To introduce the management experience in the first cause of living-related small bowel transplantation in China. Methods　An 18-year-old male patient with short gut syndrome received a living-related small bowel transplantation with the graft taken from his father(44-year-old). A segment of 150?!cm distal ileum was resected from the donor. Treatment of immunosuppression, antibiotics, antithrombosis and nutrition support were given posttransplantatively. Results　Recently the recipient has a good life quality for 19 months. Conclusions　Living-related small bowel trnasplantation can be effectively used to treat short gut syndrome, and the posttransplantative management is the key to the successful transplantation.

Objective To introduce the management experience in the first cause of living related small bowel transplantation in China. Methods An 18 year old male patient with short gut syndrome received a living related small bowel transplantation with the graft taken from his father(44 year old). A segment of 150?cm distal ileum was resected from the donor. Treatment of immunosuppression, antibiotics, antithrombosis and nutrition support were given posttransplantatively. Results Recently the recipient has a good life quality for 19 months. Conclusions Living related small bowel trnasplantation can be effectively used to treat short gut syndrome, and the posttransplantative management is the key to the successful transplantation.

Objective: To analyze the mid-term clinical efficacy of extended to C1, 2 posterior single-door laminoplasty for the treatment of ossification of the posterior longitudinal ligament combined the upper cervical spine. Methods: From February 2013 to December 2015, 32 patients with cervical OPLL who were extended to C1, 2 posterior open-door laminoplasty were enrolled in our hospital (complete follow-up of 25 patients). There were 19 males and 6 females, aged 48-76 years (mean 60.1±7.0 years). Cervical spinal cord function was assessed using the Japanese Orthopaedic Association (JOA) spinal cord function score, and the rate of improvement in neurological function was calculated. The quality of life of patients was assessed using the neck disability index (NDI). Axial symptoms were assessed and indexed using the visual analog scale (VAS). The C0-2 angle of the cervical spine was measured by X-ray preoperative and at the last follow-up. The cervical curvature was evaluated by the C2-7 angle, and the cervical vertebra activity was observed by the dynamic X-ray of the cervical spine. Results: 25 patients were followed up for a period of 26-64 months (mean 35.9±8.1 months). At the last follow-up, the JOA score was 14.32±3.24, the NDI score was 7.61±1.23, and the VAS score was 1.42±0.78. The differences between the three scores were statistically significant compared with the preoperative scores. The neurological improvement rate at the last follow-up was 79.61%±13.23%. The preoperative C0-2 angle was 26.04°±6.28°, and the last follow-up was 24.92°±5.51°; the C2-7 angle was 19.55°±9.42° before surgery, and the C2-7 angle at last follow-up was 17.97°±8.80°. The C2-7 angle at last follow-up was slightly reduced compared with that preoperative, but the difference was not statistically significant. The preoperative cervical vertebra ROM was 35.31°±12.24°, and at the last follow-up it was 32.23°±9.65°. The ROM of cervical vertebrae at the last follow-up was slightly lower than that before surgery, but the difference was not statistically significant. Among them, the reduction of the ROM of overflexion was greater than that of the over-extension, which was the main reason for the decrease of ROM. The OPLL bone mass continued to grow at the last follow-up of 11 patients. Conclusion The extended to C1, 2 posterior single-door laminoplasty for the treatment of ossification of the posterior longitudinal ligament combined the upper cervical spine can achieve adequate spinal cord decompression, satisfactory neurological improvement, and improve the postoperative cervical curvature and activity. There was no obvious change in the curvature and activity of the cervical spine, and the axial symptoms did not increase significantly, and the clinical efficacy was positive.

OBJECTIVEThe aim of this study was to compare the safety and efficacy profiles of R-CHOP and R-fCHOP regimes in the treatment of primary gastric diffuse large B cell lymphoma (PG-DLBCL).

METHODSData of PG-DLBCL patients admitted in our hospital from March 2010 to March 2014 were collected retrospectively. Differences in gastrointestinal bleeding and perforation between the R-CHOP group and R-fCHOP group were compared. The influence of the gastrointestinal complication on subsequent treatment was also analyzed. Treatment outcome of the two groups was also compared.

RESULTSA total of 50 patients were included in this retrospective study. Forty of them were in the R-CHOP group, another ten were in the R-fCHOP group. Patients in the R-fCHOP group had a higher rate of Lugano late stage disease, and a relatively high rate of a deeper/larger ulcer. Fence occult blood test (FOBT) was positive in one (10.0%) patient in the R-fCHOP group, and 11 (31.4%) patients in the R-CHOP group, among them one had hematemesis and had to give up the subsequent chemotherapy. No perforation was observed in both groups. The response rate (RR) was 92.5% in the R-CHOP group and 90.0% in the R-fCHOP group (P > 0.05). The PFS was also comparable between the two groups (P > 0.05).

CONCLUSIONSR-fCHOP regimen has a good safety profile in patients with Lugano late stage and deep/large ulcers, who are of high risk of gastrointestinal bleeding or perforation, and also has a comparable efficacy profile when compared with the R-CHOP regimen in short-term follow-up.

Adult ; Aged ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; Doxorubicin ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; Lymphoma, Non-Hodgkin ; Male ; Middle Aged ; Prednisone ; Retrospective Studies ; Rituximab ; Stomach Neoplasms ; Treatment Outcome ; Vincristine