Highly active adherent LAK cells (A-LAK) with monocytes depleted by phenylalahine methyl ester (PME) were cultured from peripheral blood lymphocytes of patients with hepatocellular carcinoma (HCC). Results showed that A-LAK cells cultured in about 14 days had expanded better and faster than that of nonadherent LAK cells (NA-LAK) with their greatest expansion varied from 23 to 243 fold .A-LAK cells showed a trend of increase in TH cell subgroup and decrease in Ts cell sub-group as well as significant difference of TH/TS ratio. The IL-2R expression increased from 34.1%to 64.3%. A-LAK cells had a higher cytotoxicity (64.6%)than that of NA-LAK cells (42.8%).Further clinical application of A-LAK cells may improve biotherapeutic effect on HCC patients compared with that of NA-LAK cells .

BACKGROUND:Restoring the stability of the spine has become the consensus of spinal surgery. The canaloplasty technology has been continuously improved, but how can we get the good clinical effect of the canaloplasty, and the price affordable, many domestic scholars have to try al kinds of the improved operation methods. OBJECTIVE:To evaluate the clinical application value of cannulated screws fixation in canaloplasty. METHODS:From February 2011 to February 2013, total y 24 patients with spinal disease treated by canaloplasty using cannulated screw were retrospectively analyzed, of which 12 cases of cervical stenosis, 2 cases of intraspinal tumor in thoracic and 10 cases of intraspinal tumor in lumbar. Al patients were fol owed up after treatment. Postoperative CT and MRI were done in al patients. Clinical symptoms and radiographic changes were observed after treatment. The Japanese Orthopaedic Association score and the spinal canal cross-sectional area measurement were conducted in the patients with cervical stenosis between the preoperation and postoperation. Visual analog scale score was evaluated in patients who have the tumor in the thoracolumbar spine between the preoperation and postoperation. RESULTS AND CONCLUSION:Al patients had no complications such as nerve or blood vessel damage. Al patients were fol owed up 12 to 24 months. Imaging evaluation showed that internal fixator was stable without the hol ow screw loss or displacement. The bone grafting in groove reached bone fusion. There was no occurrence of lamina col apse or“re-close of door”. The Japanese Orthopaedic Association score and spinal canal cross-sectional area of patients with cervical stenosis during the fol ow-up after 12 months of treatment were significantly superior to those in preoperation (P<0.01). After 12 months of treatment, Japanese Orthopaedic Association scores showed that the excel ent rate of classification assessment was 92%. During the fol ow-up after 12 months of treatment, the visual analog scale of patients with thoracolumbar tumor improved from (8.2±1.6) points before treatment to (2.3±1.3) points at the first year after discharge (P=0.004 2). These results suggest that the application of cannulated screws in the canaloplasty can not only enhance the stability of the rear pil ar, and can improve the healing rate of osteotomy, and has the characteristics of inexpensive, easy to operate, and repair effect is good.

Objective To evaluate the clinical effects of cytokine-induced killer (CIK) cells combined with chemotherapy on the treatment of patients with advanced colorectal cancer.Methods CIK cells were prepared from 50 ml peripheral blood mononuclear cells by stimulated with IL-2,IFN-γ,anti-CD3 monoclone antibody,IL-1 for 8 d.The clinical effects and survival rate were compared between CIK cells combined with chemotherapy group and the chemotherapy group (50 patients with advanced colorectal cancer for each).T cells and NK cells of patients were tested by FCM before and after CIK cells treatment.The improvement of quality of life and toxicity of this therapy were observed.Results The percentages of CD3+,CD4+,CD8+ T cells and NK cells were (54.779±14.228) ％,(30.821±11.554) ％,(16.676±6.256) ％,(18.705±9.347) ％ before CIK cells transfusion.After transfusion,the percentages were (65.236±14.901) ％,(37.292±8.880) ％,(25.229±6.711) ％,(22.950±8.933) ％,respectively.The percentages were expanded greatly (P ＜ 0.05).The patients quality of life were improved clearly with lower toxicity.The DCR of CIK cells combined with chemotherapy group (64 ％,32/50) was higher than the chemotherapy group (40 ％,20/50) (P ＜ 0.05).The survival rate between two groups had no statistical significance (P ＞ 0.05).Conclusion Administration of CIK cells combined with chemotherapy can enhance immune function in patients with advanced colorectal cancer and improve their quality of life,and get good clinical efficacy.

The antiEGFR monoclonal antibodies cetuximab and panitumumab have been proven to be efficient in MCRC. The degree of EGFR expression (as confirmed by immunohistochemical analysis) did not correhte with the clinical response. In this review, we describe the current status of markers that might identify patients who are likely to benefit from treatment with cetuximab or panitumumab. These molecular markers include KRAS mutations, EGFR copy number, EGFR ligands (EGF, epiregulin), cyclin DI, IgG FcγR (FCGR2A-HI31R and FCGR3A-V158F), and nuclear factor-κB, that are crucial to avoid anti-EGFR treatment toxicity and reduce treatment cost.

AIM: To construct human Egr-1 promoter luciferase reporter system and study its activity induced by i-onizing radiation. METHODS: Egr-1 promoter was obtained by human genomic PCR and cloned into pGL3-basic vector. After transfection of recombinant plasmid into human tumor cells, the Egr-1 promoter activity induced by ionizing radiation was detected by luciferase reporter assay. RESULTS: The luciferasy reporter system of Egr-1 promoter was successfully constructed. The activity of Egr-1 promoter was substantially increased after different doses of IR and reached to the peak at the time point of 48h after IR. CONCLUSION: The Egr-1 promoter was constructed in this study showed IR inducible activity in tumor cells, laying foundation for the research of radiation, mediated gene therapy.

Background and purpose:Liquid biopsy is a kind of blood, urine and other non-solid biologi-cal tissue sampling analysis, mainly for malignant tumor diagnosis, monitoring and predicting its prognosis. In this research, we optimized the extraction of miRNA in urine, established a standardized means of liquid biopsy, screened and verified the miRNA markers in patients with bladder cancer.Methods:From Jan. 2014 to Sept. 2015, we used miRNA microarray in six patients with bladder cancer and six healthy controls. Samples of 78 cases of bladder cancer and 23 healthy controls were tested by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) to verify the relationship between miRNA markers in liquid biopsy and clinical pathological parameters. The diagnostic value of miRNA markers was also analyzed and compared.Results:We screened 10 miRNAs differential expression in urine. Combined with previous literature, we selected 20 miRNAs to verify their expression levels in bladder cancers and healthy controls. miR-509-5p/miR-124 ratio in the urine was found higher in patients with bladder cancer than in healthy controls (P<0.0001). With the rise of miR-509-5p/miR-124 ratio in urine, tumor stage and grade were also increased (P=0.003). When the cutoff was set at 0.41, the diagnostic sensitivity and specificity of miR-509-5p/miR-124 ratio were 73.1% and 82.6%, respectively. The AUC of miR-509-5p/miR-124 ratio to detect bladder cancer was 0.864, higher than that of urinary exfoliated cells (P=0.0002).Conclusion:We optimized the extraction of miRNAs in urine,established a standardized liquid biopsy of miRNA markers. The miR-509-5p/miR-124 ratio could be an ideal diagnos-tic marker for bladder cancer.

Objective:To investigate the prognostic value of gastroepiploic lymph node (GLN) metastasis in transverse colon cancer.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 371 patients with transverse colon cancer who were admitted to Fujian Medical University Union Hospital from November 2010 to November 2017 were collected. There were 202 males and 169 females, aged from 21 to 92 years, with a median age of 58 years. Patients were performed complete mesocolic excision combined with GLN dissection by one group of surgeons. Of the 371 patients with transverse colon cancer, 15 cases had positive GLN metastasis (GLN+), and 356 cases had negative GLN metastasis (GLN-). Observation indicators: (1) the propensity score matching conditions and comparison of baseline data between GLN- patients and GLN+patients with transverse colon cancer after propensity score matching; (2) follow-up and survival of GLN- patients and GLN+patients with transverse colon cancer; (3) influencing factors for prognosis of patients with transverse colon cancer. Patients were followed up by outpatient examination or telephone interview to detect tumor metastasis and survival. Follow-up was conducted once every 3 months within postoperative 2 years, once every 6 months within postoperative 2-5 years and once a year thereafter up to January 2020. The propensity score matching was conducted by 1∶4 matching using the nearest neighbor method. Measurement data with skewed distribution were described as M (range), and comparison between groups was analyzed using the rank sum test. Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. The Kaplan-Meier method was used to calculate survival rates and draw survival curves, and Log-rank test was used for survival analysis. Univariate and multivariate analyses were performed using the COX proportional hazard regression model. The variables with P<0.10 in the univariate analysis were included for multivariate analysis. Results:(1) The propensity score matching conditions and comparison of baseline data between GLN- patients and GLN+ patients with transverse colon cancer after propensity score matching: 55 of 371 patients had successful matching, including 44 GLN- patients and 11 GLN+ patients. Before propensity score matching, the age, cases in stage 0 or stage 1 of M staging, preoperative carcinoembryonic antigen were 60 years(range, 24-92 years), 328, 22, 4.1 μg/L(range, 0.2-343.7 μg/L) for GLN- patients, respectively, versus 67 years(range, 21-79 years), 11, 4, 5.0 μg/L(range, 0.7-952.4 μg/L) for GLN+ patients, showing significant differences in the above indicators between the two groups ( Z=-1.440, χ2=9.031, Z=-2.086, P<0.05). After propensity score matching, the above indicators were 58 years(range, 45-67 years), 40, 4, 4.0 μg/L(range, 2.0-10.0 μg/L) for GLN- patients, respectively, versus 67 years(range, 59-71 years), 9, 2, 5.0 μg/L(range, 8.0-19.0 μg/L) for GLN+ patients, showing no significant difference between the two groups ( Z=-1.580, χ2=0.105, Z=-0.821, P>0.05). (2) Follow-up and survival of GLN- patients and GLN+ patients with transverse colon cancer: GLN- patients and GLN+ patients with transverse colon cancer were followed-up for 12-92 months and 1-70 months, with a median time of 53 months and 30 months respectively. Three cases of GLN- patients and 2 cases of GLN+patients had postoperative liver metastasis, respectively, showing no significant difference between the two groups ( χ2 =0.344, P>0.05). One case of GLN- patients and 3 cases of GLN+ patients had heterochronous lung metastasis, respectively, showing a significant difference between the two groups ( χ2 =4.870, P<0.05). The 5-year disease progression-free survival rates were 82.3% and 33.9% for GLN- patients and GLN+ patients, respectively, showing a significant difference between the two groups ( χ2 =13.366, P<0.05). (3) Influencing factors for prognosis of patients with transverse colon cancer: results of univariate analysis showed that pT staging, pN staging, M staging and GLN metastasis were related factors for prognosis of patients with transverse colon cancer ( hazard ratio=1.599, 5.107, 4.511, 6.273, 95% confidence interval as 0.467-5.471, 1.867-13.971, 1.385-14.694, 2.052-19.176, P<0.05). Results of multivariate analysis showed that pN staging, M staging and GLN metastasis were independent influencing factors for prognosis of patients with transverse colon cancer ( hazard ratio=6.399, 6.163, 4.024, 95% confidence interval as 2.028-20.189, 1.666-22.800, 1.177-13.752, P<0.05). Conclusion:For the patients with transverse colon cancer, GLN metastasis is associated with high postoperative heterochronous lung metastasis rate and poor prognosis. GLN metastasis is an independent prognostic factor for patients with transverse colon cancer.

Background and purpose: Renal-cell carcinoma (RCC) is susceptible to immune therapy including the use of the nonmyeloablative allogeneic transplantation(NAT). However, NST can produce severe toxicity, so it might not be appropriate for many patients with metastatic RCC. Other novel allogeneic immunotherapies have been designed to induce an autologous immune response directed against the malignancy. This study evaluated the efficacy and safety of infusions of partially HLA-matched irradiated allogeneic blood mononuclear cells for advanced renal-cell carcinoma. Methods: Patients with histologically proven diagnosis of advanced RCC received infusions of partially HLA-matched allogeneic blood mononuclear cells. Repeat infusions were given every 8 weeks. Treatment was continued until disease progressed, unacceptable toxicity, or patient (or donor) choice. Results: Eight patients were enrolled. After every infusion, 6 patients received an oral administration of thalidomide daily with 100-300 mg/d for 2 months. One patient had durable complete response. Five stable diseases and two progress diseases were observed. In eight patients, time to progression and survival were 320 and 879+days, respectively. Severe toxicity was not observed. Conclusion: Infusions of partially HLA-matcbed irradiated allogeneic blood mononuclear cells for advanced RCC may induce some antitumor effects and deserves further study.

Objective To construct the prokaryotic expression vector of human autophagy-related LC3B gene,obtain the GST-LC3B recombinant plasmid , purify the GST-LC3B fusion protein and identify its activity in vitro.Methods Human LC3B coding region was amplified from the human mammary gland cDNA by PCR and inserted into the prokaryotic expres -sion vector pGEX-KG.The recombinant plasmid pGEX-KG-LC3B was transformed into E.coli Rossate.The expressed product was purified by GST-Sepharose 4B beads and identified by SDS-PAGE and Western blot analysis .The function of the purified protein GST-LC3B was detected by GST pull-down assay.Results About 400 bp of the LC3B coding region was successfully amplified from the mammary gland library by PCR and inserted into pGEX -KG.The result of double diges-tion and sequencing showed that the GST-LC3B recombinant plasmid was successfully obtained .The GST-LC3B fusion pro-tein of about 40 000 (Mr) was successfully purified and identified by SDS-PAGE and Western blotting analysis.GST pull-down assay showed that GST-LC3B could interact with Atg4B, which identified its known function .Conclusion The pro-karyotic expression vector of GST-LC3B is constructed successfully , which will facilitate further research on the function of LC3B in autophagy.

Objective To explore the change and significance of blood lipid and oxidative stress in subchnical hypothyroidism during pregnancy.Methods 100 pregnant women with subclinical hypothyroidism during pregnancy were chosen and set as the disease group,while 100 healthy pregnant women were chosen and set as the control group.Thyroid function index:free three iodine thyroid original glycine (FT3),free thyroid hormones (FT4),thyroid stimulating hormone (TSH),blood lipids index:triglyceride (TG),total cholesterol (TC),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C),and oxidative stress indicators:superoxide dismutase (SOD),malondialdehyde (MDA),type oxidized low density hpoprotein (ox-LDL) in the two groups were detected and analyzed.Results FT3 and FT4 between the two groups had no statistical significance (P＞0.05).TSH in the disease group was (4.63±1.09) mIU/L,significantly higher than that of the control group(P＜0.05).The comparison of TG,HDL-C between the two groups had no statistical significance(P＞0.05).TC and LDLC in the disease group were significantly higher than those of the control group (P＜0.05).MDA and ox-LDL in the disease group were significantly higher than those of the control group,while SOD was significanTLy lower than that of the control group(P＜0.05).Conclusion Subclinical hypothyroidism during pregnancy can cause metabolic disorder of blood lipid and reduce oxidative stress ability.Monitoring and intervention of blood lipid and oxidative stress status is necessary.