Objective To explore the clinical treatment of patients with stage Ⅲ carcinoma of colon and rectum by tegafur chemotherapeutic drugs combined with standardized adjuvant therapy. Methods The data of 60 cases of carcinoma of colon and rectum treated in Wenzhou Central hospital from January 2013 to December 2015 were collected. The patients were divided into three groups, non standardized adjuvant treatment group of 18 cases, standardized adjuvant treatment group of 15 cases and tegafur drug combined with standardized treatment group of 27 cases. Recorded and compared the survival rate of the three groups within three years, the statistics of each group of disease-free survival (DFS) and overall survival rate (OS). According to the duration of treatment group, the use of oxaliplatin combined with 5-fluorouracil and leucovorin (FOLFOX) treatment, analyzed and recorded OS and DFS. Follow-up examinations were performed in patients who did not receive adjuvant therapy. Recorded and compared the OS and DFS of each group. Results Tegafur drug combined with standardized treatment group DFS and OS were significantly higher than the other two groups, the difference was statistically significant (P<0.05). In addition, when treated with FOLFOX regimen, the DFS and OS of group Ⅲ were significantly higher than those of groupⅠand groupⅡ, the difference was statistically significant (P<0.05). In the follow-up survey of three groups, the rates of recurrence and death in patients treated with tegafur drug combined with standardized treatment group were significantly lower than those in the non standardized adjuvant treatment group and the standardized adjuvant treatment group. Conclusion Tegafur combined with adjuvant therapy can significantly improve the standardization of the rate of carcinoma of colon and rectum and the rate of disease-free survival, it is suggested that doctors use chemotherapy combined with auxiliary standardized therapy, can significantly improve the survival of patients, and reduce the risk of disease recurrence rate.

Background and purpose: Multidrug resistance (MDR) of tumor cell is the main reason for clinical chemotherapy failure as well as cancer recurrence and metastasis. This study was to construct a lentiveral vector of RNA interference of MDR1 gene and observe its inhibitive role on the expression of MDR1 in A549 and A549/DDP cells. Methods: Oligos of base pairs for hairpin RNA targeting MDR1 were chemically synthesized. Via annealing and inserting them into the down-stream of H1 promoter of linearized pSUPER, we obtained the siRNA constructs for MDR1, which were afterwards transfected into A549, a human lung cancer cell line expressing high level MDR1, the impact of constructs was observed on the expression and interference efficiency of siRNA against MDR1. The effective sequence of siRNA targeting MDR1 gene was confirmed. Both sense and antisence oligo DNA of the targeting sequence was designed, synthesized and cloned into the PTM vector, containing a promoter and a green fluorescent protein (GFP). The resulting lentiviral vector containing MDR1 siRNA was named PTM-siMDR1 and then transfected into A549 and A549/DDP cells after being confirmed by PCR and sequencing. Results: Restriction digestion and DNA sequencing showed that the siRNA constructs for MDR1 were successfully produced and the expressed siRNA could effectively down-regnlate the expression of MDRI. PCR demonstrated that the lentivirus RNAi vector of MDR1 producing PTM-siMDR1 was constructed successfully. The chemosensitivity of A549/DDP cells to cisplatin were enhanced obviously after trartsfection. Conclusion: The lentivirus RNAi vector of MDR1 can significantly revise the resistance ofA549/ DDP cells with eisplatin after infection.

Objective:To investigate the clinical features, risk factors, treatment and prognosis of dermatomyositis (DM) patients with positive anti-melanoma differentiation associated gene 5(MDA5) antibody with rapidly progressive interstitial lung disease (RPILD).Methods:The clinical data of 88 DM patients from June 2019 to June 2020, at the rheumatology department of Guangdong Provincial People's Hospital were collected and retrospectively analyzed. T-test, non-parametric Mann-Whitney U test, Chi-squared test, Fisher exact probability and Logistics regression analysis were used for data analysis. Results:① 37%(36/88) DM patients were positive for anti-MDA5 antibody. The frequency of ulcerative rash, Gottron's sign, arthritis, clinically amyopathic dermatomyositis (CADM), and erythrocyte sedimentation rate (ESR) was significantly higher in patients with anti-MDA5 antibody ( P<0.05). The cell count of white blood cell, neutrophil, lymphocyte, and serum creatine kinase (CK) level were significantly lower in the anti-MDA5 antibody positive group than those in the negative group ( P<0.05). Of anti-MDA5 antibody positive DM patients, 100% developed ILD, 34% (11/32)developed RP-ILD, 16%(5/32) died, which were significantly higher than those of anti-MDA5 antibody negative patients ( P<0.05). ② Of anti-MDA5 antibody positive DM patients, the C reactive protein (CRP) level, positive rate of anti-Ro-52 antibody and mortality rate were significantly higher RPILD group than those in the non-RPILD group [15.70(4.49, 29.00) vs 3.22 (1.66, 7.15), Z=-2.440, P=0.014; 91% vs 43%, P=0.011; 46% vs 0, P=0.002]. Logistics regression analysis indicated that positive anti-Ro-52 antibody [ OR=4.561, 95% CI (1.797, 11.580), P=0.001] might be a risk factor for anti-MDA5 antibody positive DM-RPILD. ③ Among patients with anti-MDA5 antibody with RPILD, serum ferritin and D-dimer level was significantly higher and oxygenation index was significantly lower in the non-survival group than those in the survival group [1 931 (1 377, 7 379) vs 638(196, 876), Z=-2.556, P=0.009; 2 760(1 995, 4 854) vs 985(533, 1 588), Z=-2.379, P=0.017; 230(140, 256) vs 309(262, 382), Z=2.191, P=0.030]. In addition, the delayed intensive treatment time was significantly longer in the non-survival group than those in the survival group [(14.0±2.6) vs (4.5±1.4), t=7.899, P<0.01]. Furthermore, the proportion of combined therapy with two disease modifying antirheumatic drug (DMARDs) was significantly lower in the non-survival group than those in the survival group (0 vs 83%, P=0.015). Conclusion:Anti-MDA5 antibody may be associ-ated with characteristic clinical manifestations of DM, ILD, RPILD and high mortality rate. Positive anti-Ro-52 antibody may be a risk factor for anti-MDA5 antibody positive DM-RPILD. High serum ferritin and D-dimer level and low oxygenation index in RPILD patients may be associated with poor prognosis. Early treatment with two DMARDs may improve the prognosis of RPILD.

Objective: To determine the extent of transfection and expression of adeno-associated virus (AAV) serotype 9 (AAV9) in the cochleae of mice at different ages. Methods: AAV9-green fluorescent protein (GFP) was inoculated into the cochlea of mice via the round window membrane (RWM) or through cochleostomy at different ages. Four groups were divided according to ages and injection sites: P1SM group, AAV9-GFP was delivered to the scala media by cochleostomy at postnatal day 1; P1RW group, AAV9-GFP was delivered to the scala tympani via RWM at postnatal day 1; P9RW group: AAV9-GFP was injected through RWM at postnatal day 9; and P30RW group, adult mice (P30) were injected through RWM. GFP expression in cochlear whole mount was analyzed and auditory brainstem response (ABR) tests were conducted one month after virus injection (for each animal, only left cochlea was injected and the right side was used as a control). GraphPad Prism 5 statistical software was used for data analysis. Results: All of inner hair cells (IHCs) and most of outer hair cells (OHCs) were transfected via two approaches at P1 injection. There was no significant difference in ABR threshold between injected ears and untreated ears (P>0.05). All of the IHCs and parts of OHCs (69% in apical turn) were transfected via RWM at P9. The strongest GFP expression was observed near the apical turn. Cochlear inoculation via RWM at P30 led to transgene expression in only IHCs. The ABR threshold of injected ears in P9RW group and P30RW group was significantly higher than that of contralateral ears (P<0.01). Conclusions AAV9 can be highly expressed in the inner and outer hair cells of the cochlea and hearing sensitivity can be preserved if virus injections are performed in neonatal mice. After AAV9 is transfected into the inner ear of adult mice, it is only expressed in the inner hair cells, which leads to the increase of the ABR response threshold of mice. Transfection efficiency is significant higher in neonate mice than in P9 and adult mice.

Objective: To investigate the number and distribution of dendritic cells (DCs), macrophages and M2 macrophages in renal tissues of patients with IgA nephropathy (IgAN) and their correlation with clinicopathological parameters, and explore its role in the progression of IgAN. Methods: Renal tissue samples from 42 patients aged ≥18 years with IgAN were collected by kidney biopsy in Guangdong Provincial People's Hospital from January 2018 to June 2018. The patients were divided into different groups according to Oxford classification and Lee grade classification criteria. The distribution and number of DCs (CD209), macrophages (CD68) and M2 macrophages (CD68 and CD206) were detected by immunohistochemistry. Spearman correlation test was used to analyze the correlation between the number of DCs and macrophages in renal tissues and clinical pathological parameters. Results: The number of DCs in the glomeruli of the M1, T0 and C1 groups increased significantly compared with the M0, T1 and C0 groups, and the number of DCs in the renal interstitium of the T1 group increased significantly compared with the T0 group (all P<0.05). The number of glomerular macrophages in group C1 was significantly higher than that in group C0. The number of macrophages in S1, T1 and Lee IV-V tubulointerstitial groups was significantly higher than that in S0, T0 and Lee II-III groups (P<0.05). The number of M2 macrophages in the S1, T1 and Lee IV-V groups was significantly higher in the tubulointerstitial group than in the S0, T0 and Lee II-III groups (all P<0.05). The blood urea nitrogen, serum creatinine and 24 h urine protein levels in the T1 and Lee IV-V groups were significantly higher than those in the T0 and Lee II-III groups, and the serum albumin levels in the Lee IV-V group were significantly lower (all P<0.05). Spearman correlation analysis showed that the number of DCs in renal interstitium was positively correlated with the proportion of tubulointerstitial fibrosis. There were a positive correlation between the proportion of glomerular segmental sclerosis and tubulointerstitial fibrosis and renal interstitial macrophages and M2 macrophages. The number of M2 macrophages was positively correlated with serum creatinine and 24 h urine protein (both P<0.05). Conclusions The number of DCs and M2 macrophages in kidneys are positively correlated with the clinicopathological features of IgAN patients, which indicates that they may be associated with disease progression.