Objective To study the clinical characteristics of very elderly patients aged 80 years and over with diabetes mellitus (DM).Methods Clinical data of 95 very elderly patients with diabetes mellitus were retrospectively analyzed.Results Among 95 patients,patients with type 2 diabetes mellitus accounted for 98.9％ (94/95),patients with asymptomatic onset accounted for 35.8％ (34/95).The incidence of acute complications of diabetes mellitus was 11.6 ％ (11/95).Chronic complications of peripheral neuropathy and diabetic nephropathy were more common,and their incidences were 83.2％ (79/95) and 45.3％ (43/95),respectively.The percentage of patients with multiple chronic complications was up to 45.3％ (43/95).The percentage of DM patients combined with hypertension,coronary heart disease,cerebral infarction,hyperlipidemia or cardiac valve calcification was 80.0％ (76/95),48.4％ (46/95),77.9％ (74/95),42.1％ (40/95)and 33.7％ (32/ 95),respectively.There were 29 patients (30.5％) with the simultaneous presence of DM,hypertension,cerebral infarction,coronary heart disease,and hyperlipidemia in a same patient.The proportion of DM patients with low serum albumin was 47.4％ (45/95),and the rate achieving the target low density lipoprotein level was only 35.8％ (34/95).In the treatment,67.4％ (64/95) of patients were treated with oral hypoglycemic agents combined with insulin injection.The incidence of hypoglycemia was 24.2％ (23/95),and 69.6％ (16/23) of them had no self-conscious hypoglycemic symptoms.Conclusions Chronic complications are common in elderly diabetes mellitus patients.The elderly DM patients are prone to have many complications at the same time,with a higher incidence of hypoglycemic value under 3.9 mmol/L and lower rate of hypoglycemic symptoms.

Objective To establish a model to predict the clinical response of neoadjuvant chemotherapy for nasopharyngeal car‐cinoma ,and provide basis for the individual treatment .Methods The clinical data of 63 cases of advanced nasopharyngeal carcinoma patients who have received neoadjuvant chemotherapy in the past 2 years were analyzed retrospectively .Univariate and multivariate analyses were performed using the Logistic analyses to identify efficacy factors .Results The response rate in nasopharyngeal tumor and lymph node metastasis were 39 .7% and 50 .8% ,respectively .Single factor analysis showed that patients with no distant metas‐tasis ,cranial nerve inviolated ,EBV negative and high expression of Ki67 were more sensitive to therapy .Logistic analysis showed that the influencing factors for the effect of the new chemotherapy include :distant metastasis ,cranial nerve inviolated and EBV . Thus ,the prediction model would be:Logit= -0 .470 -2 .863 × distant metastasis + 1 .328 × cranial nerve invasion+ 3 .639 × EBV ,its sensitivity ,specificity ,positive predictive value and negative predictive value were 79 .4% ,82 .8% ,84 .4% and 77 .4% . Conclusion The distant metastasis ,cranial nerve invasion and EBV infection were important predictive factors for neoadjuvant chemotherapy of nasopharyngeal carcinoma .This model could be used to predict the response of patients with nasopharyngeal carci‐noma .

Objective To evaluate the curative effect and optimal indications of reformed percutaneous lumbar diskectomy (RPLD) for the treatment of lumbar disc herniation.Methods One handred and thirty-three patients with lumbar disc herniation were treated by RPLD, of them, 20 cases were lumbar disc extrusion and 113 cases were lumbar disc protrusion. After the procedure, 85 patients underwent flush of intervertebral space with antibiotic saline and 48 patients underwent 10 ml(40?g/ml) medical ozone-injection inside the disc to prevent infection. All patients were followed up over the course of 3 months. The therapeutic effect was evaluated according to the MacNab criteria. Results All 133 cases underwent RPLD were successful. The total efficacy was 81.9%, There were no serious complications after operations. Conclusion RPLD is an effective method in the treatment of lumbar disc herniation. Both intradiscal ozone-injection or intradiscal antibiotic saline flush after RPLD can reduce the opportunity of infection.

Objective To understand the therapeutic effect of clindamycin combined with compound sulfamethoxazole tablets on pneumocystis pneumonia(PCP)associated with acquired immunodeficiency syndrome (AIDS).Methods 97 AIDS patients with PCP in a hospital from January 2014 to March 2015 were randomly divided into control group (n=49,received compound sulfamethoxazole )and trial group(n=48,received clindamycin on the basis of com-pound sulfamethoxazole ),levels of partial pressure of oxygen in arterial blood (PaO2 ),arterial blood oxygen satu-ration(SaO2 ),serum albumin(ALB),and lactic dehydrogenase (LDH)in two groups of patients before and after treatment were recorded.Results Levels of PaO2 ,SaO2 ,ALB,and LDH between two groups of patients before treatment was not significantly different(all P >0.05).After treatment,PaO2 in control group and trial group were (73.01 ±4.62)mmHg and(84.92 ±5.34)mmHg respectively,SaO2 were (75.81 ±4.28)% and(90.86 ±5.94)%respectively,ALB were (32.62±4.41 )g/L and(43.95 ±5.03)g/L respectively,LDH were(416.53 ±30.77)U/L and(331 .58±20.86)U/L respectively,levels of PaO2 and SaO2 in trial group were both higher than control group , difference in ALB and LDH between two groups of patients after treatment were both statistically significant(both P <0.05).The total effective rate of trial group was 89.58% (n=43),which was higher than 69.39%(n=34)in control group (χ2 =6.04,P =0.014).Conclusion Clindamycin combined with compound sulfamethoxazole tablets has good therapeutic effect on AIDS and PCP,which is worthy of clinical popularization and application.

Objective To detect the mutations of GJB2 and GJB6 genes in the first Chinese case of keratitis, ichthyosis and deafness (KID) syndrome. Methods Genomic DNA was extracted from the patient with KID syndrome and his family members. All encoding exons and adjacent splice sites of the GJB2 and GJB6 genes were amplified by PCR. Mutation scanning was carried out by direct bidirectional DNA sequencing. Results No mutation was found in GJB6. A G148A mutation was found at exon2 of GJB2 in the patient, which caused a change from aspartic acid to asparagine at codon 50(D50N). Conclusion This case of KID syndrome may be caused by the mutation in GJB2.

Objective To evaluate the feasibility of monitoring the gene expression of VEGF165 via the diglycylcysteine (GGC) reporter gene system by reporter probe of 99Tcm-GH. Methods DNA fragments encoding GGC binding motifs were prepared by PCR and positioned at the C end of VEGF165 gene after the linearization of pcDNA3-VEGF165 plasmid. A replication-defective adenovirus vector Ad5-VEGF165GGC motif-internal ribosomal entry site(IRES) -enhanced green fluorescent protein (EGFP) (Ad5-VIE)was constructed, with a cytomegalovirus (CMV) early promoter driving the expression of VEGF165 gene,GGC motif and EGFP, under the aid of an IRFS. A replication-defective adenovirus carrying the Ad5-EGFP was used as the control. Mensenchymal stem cells (MSC) were infected with the recombinant adenovirus at a multiplicity of infection (MOI) from 0 to 100 infectious units (0,10,25,50,100). The cellular uptake of 99Tcm-GH in infected MSC were then studied at 30, 60, 90 and 120 min. VEGF165 was detected by quantitative reverse transcriptase real-time PCR (RT-PCR), Western-blot, and immunohistochemisty. EGFP was observed by RT-PCR and fluorescence microscopy. The correlation analysis was studied between the cellular uptake of 99Tcm-GH and the expression of VEGF165. SPSS 13.0 was applied for statistical analysis. Independent samples t-test, q-test and Pearson correlation coefficient were used. Results After infected with different viral titer of Ad-VIE, the cellular uptake of 99Tcm-GH increased with the increasing virus titer(r2 =0.86, P ＜0.05), with the peak rate (7.94 ±0.75) % at MOI = 100. In time-dependent uptake study, the cellular uptake rates increased rapidly with the time extension, and the highest uptake occurred at 120 min with the peak uptake rate (7.72 ±0.22)%. The uptake rates of 99Tcm-GH in Ad5-VIE-infected cells were significantly higher than those of Ad5 -EGFP-transfected cells at all time points (t = 15.10- 54.92, all P ＜0.05). The VEGF165 and EGFP mRNA levels increased with increasing virus titer, and the VEGF165 mRNA correlated well with the EGFP mRNA(r2 = 0. 99, P ＜ 0.05). After infected with different MOI of Ad5-VIE, good relationship was found between the cellular uptake of 99Tcm-GH and the expression of VEGF165protein in MSC(r2 =0.90, P ＜0.05). Inmunohistochemisty showed VEGF165 protein expressed obviously at Ad5-VIE-infected MSC, and the EGFP was observed by fluorescence microscopy. Conclusions The cellular uptake of 99Tcm-GH in Ad5-VIE-infected MSC are well correlated with the expression of VEGF165 in vitro. The expression of therapeutic gene VEGF165 can be monitored by the GGC peptide expression.

Objective To explore the effects and mechanism of bexarotene in combination with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on apoptosis of leukemic cell line KG1a. Methods KG1a cells at logarithmic growth phase were obtained, and were divided into TRAIL group, bexarotene group, 300 ng/mL TRAIL in combination with bexarotene group and 2.0 μmol/L bexaroten in combination with TRAIL group. Cell apoptosis rate was detected in each group by flow cytometry. Flow cytometry was also employed to determine the apoptosis rates of KG1a cells after treatment with bexarotene and TRAIL in different sequences. The expression of Fas associated death domain-like IL-1 beta converting enzyme inhibitory protein (c-FLIP) was detected by Western blotting. Results There was no significant difference in cell apoptosis rates between TRAIL group and bexarotene group of each concentration (except for bexarotene 2.0 μmol/L) (P > 0.05). The cell apoptosis rates of 300 ng/mL TRAIL in combination with bexarotene group and 2.0 μmol/L bexaroten in combination with TRAIL group were significantly higher than those in TRAIL group and bexarotene group of each corresponding concentration (P <0.01). Sequential analysis revealed that bexarotene could reverse the resistance of KG1a cells to TRAIL (P < 0.001). Compared with single use of 2.0 μmol/L bexarotene or 300 ng/mL TRAIL, combination use could significantly down-regulated the expression of c-FLIP (P < 0.05). Conclusion Bexarotene can significantly enhance the apoptosis of KG1a cells induced by TRAIL, which may be attributed to the down-regulation of c-FLIP expression.

Objective To determine the influence of protein fusion on the biological characteris- tics of hymidine kinase(TK)and human immunodeficiency virus(HIV)Tat recombinant protein. Methods By utilizing polymerase chain reaction(PCR)technique,different fragments containing two,four or six glycines(Gly)were inserted between the HIV Tat gene and TK,and cloned into PBK vector.After testified by sequencing,the vectors were transfected into E coli.After induced by iso- propyl thiogalactose(IPTG),bacilli were collected and destructed by ultrasonic,the fusion proteins were determined by monoclonal antibody against HIV protein.HepG2 cells were incubated in DMEM supplement with 10?g/mL HIV-Gly(n)-TK(n=0,2,4,6)fusion protein,TK-HIV Tat and only HIV Tat.HepG2 cells in different groups were detected by immunofluorescence assay 24 hours after transduction with HIV Tat monoclonal antibody.The rate of apoptosis after cells were incubated with gencilovir(10?g/mL)for 3 days was determined by cell flow cytometry,while survival cell ratio was recorded by trypan blue.The data were analyzed by statistics(t-test).Results The Tat-Gly(n)-TK (n= 0,2,4,6)recombinant genes were constructed and inserted into PBK vectors,which were expressed in E coli and then purified.Cells in different groups,which were incubated with Tat-Gly (n)-TK(n=0,2,4,6)fusion proteins,Tat-TK fusion protein,TK-Tat fusion proteins or only Tat proteins respectively,were detected by immunofluorescence assay.The intensities of fluorescence in different groups were almost same,but the ratios of cell survival or apoptosis were different.The highest ratio of cells apoptosis(14.77%)was in the group that cellular culture medium was mixed with Tat-Gly(4)-TK fusion protein,followed by the groups containing 6,2 glycines or no TK gene in genes(4.30%,12.69% and 1.03%,respectively).There were significant differences between each 2 groups among the all groups(t-test,P

Plant bioreactor called mocular farming has enormous potential to produce recombinant protein infinitely. Products expressed in plants has nature physico-chemical property and bioactivity. Plant bioreactor could be an safe, economic and convenient production system which has been widely applied in industries and agriculture, especially in the life science and pharmaceutical industry. The application of recombinant transgenic plant in the production of vaccines, antibodies and pharmaceutical proteins has become a hot point in the plant genetic engineering both at home and broad. However, there are some limiting factors of application such as yield, downstream processing and so on. The advantages and research progress for the mocular farming of pharmaceutical proteins recent three years was discussed, focusing on the existing problems and new strategies in this area.

BackgroundThe immunotherapy for retinoblastoma(RB) is gradually concerned recent year.To seek relative immune-associated antigen is a basis of immunotherapy.NY-ESO-1 and NY-SAR-35 are two kinds of genes of cancer testis antigen( CTA ).To understand their expressions in RB tissue can offer index for relative study.ObjectiveThis study was to investigate the expressions of two CTA NY-ESO-1 and NY-SAR-35 in RB and explore the possibility of them as potentially promising targets for antigen-specific immunotherapy of RB.Methods The samples were obtained from 15 RB eyes,12 non-tumor retinopathy eyes and 22 normal eyes with other benign eye diseases.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expressions of NY-ESO-1 mRNA and NY-SAR-35 mRNA in the samples.Genes of positive PCR results were sequenced randomly.The relevance of the expression of the two cancer-testis antigen genes with the clinical characteristics such as tumor stage,tumor size and clinical stage were analyzed.This study was approved by Ethic Committee of Guangxi Medical University.Written informed consent was obtained from each patient before operation. Results NY-ESO-1 mRNA was positively expressed in 6 RB samples and NY-SAR-35 mRNA was expressed in 9 RB samples.In the non-tumor retinopathy samples and normal eye tissues,NY-ESO-1 mRNA and NY-SAR-35 mRNA were absent.No significant relevances were found between the expressions of the NY-ESO-1 mRNA and NY-SAR-35 mRNA with clinical characteristics such as age ( P =0.426,0.822 ),gender ( P =0.180,0.464 ),pathological classification ( P =0.744,0.582 ),tumor size ( P =0.760,0.790),and clinical stage ( P =0.868,0.707 ).Conclusions NY-ESO-1 and NY-SAR-35 have high expressing frequencies in RB tissue and their expressions in RB have specificity.These results offer a clue for the identification of targets antigen of RB.