【Abstract】ObjectiveTo investigate the expression of Tob mRNA in human colorectal cancer tissues, and their corresponding paracancerous normal tissues which was 10 cm above the tumor and pathologically proved and to explore the role of Tob mRNA in the pathogenesis of colorectal cancer. MethodsQuantitative real time RTPCR was used to detect the expression of Tob mRNA in 31 colorectal cancers. ResultsCompared with paracancerous tissue, the expression of Tob mRNA in colorectal cancer tissues was significantly increased. Moreover, the expression levels of Tob in Dukes A, B, C, D were 1.146±0.067, 1.120±0.073, 1.052±0.020 and 1.047±0.010 respectively. Analyzed by oneway ANOVA, there were significant differences in expression of Tob in different Dukes stage. ConclusionThe upregulation expression of Tob mRNA may be closely associated with tumorigenesis of colorectal carcinoma.
Objective T o observe the efficacy and safety of intravitreal injection of ranibizumab (Lucentis) for 12 consecutive months treating exudative age-related macular degeneration (EAMD). Methods This is an open and prospective study without control trial. Twenty-two eyes from 22 patients (18 males and 4 females) with EAMD diagnosed by fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA) were enrolled in this study. The patients aged from 46 to 79 years with the mean of 68.2plusmn;9.3 years. All of the patients received intravitreal injection of ranibizumab (0.5 mg, 0.05 ml) once a month for 12 consecutive months. The best-corrected visual acuity (BCVA) was obtained using the international standard visual acuity chart (converted into logMAR for statistical analysis). Central retinal thickness (CRT) was measured by optical coherence tomography (OCT) before and after each monthly treatment. Intraocular pressure (IOP) was measured using non-contact tonometry before treatment and 1 hour and 1 day after treatment. FFA and ICGA were performed every 3 months. The BCVA was 0.01 -0.9 with the average of 0.26plusmn;0.22. The average logMAR BCVA was 0.76plusmn;0.44, the CRT was 182-559 mu;m with the mean of (302.62plusmn;90.18) mu;m, and the IOP was normal before treatment. Only 13 of 22 patients completed 12 months of follow-up. The BCVA, CRT and IOP after treatment were compared with baseline using paired t test. Results Of all the 22 patients, the mean logMAR BCVA at 1 and 3 months after treatment were 0.52plusmn;0.32 and 0.37plusmn;0.27 respectively, both of which were significantly different compared with before treatment (t=4.518, 6.237;P＜0.05). Of the 13 patients, the mean logMAR BCVA at 1, 3 and 12 months after treatment was 0.51plusmn;0.34, 0.35plusmn;0.26, 0.34plusmn;0.30 respectively. All of these measures were significantly different compared with 0.74plusmn;0.37 before treatment (t=3.443, 5.438, 4.756;P＜0.05) . The mean CRT at 1, 3 and 12 months after treatment were (228.85plusmn;54.93), (231.00plusmn;38.94) and (262.92plusmn;70.48) mu;m. There were significant differences among before and 1, 3 months after treatment (t=2.914, 3.199;P＜0.05), but not between before and 12 months after treatment (t=1.408, P＞0.05). The first 3 injections contributed to the most BCVA gain with 6 patientsprime; BCVA increasing from 0.1- 0.5 to 0.5 or above. The greatest CRT reduction was obtained at 1 month after the first injection. IOP increased 1 hour after treatment and recovered within 1 day. No intravitreal injection-related side effects such as endophthalmitis were observed during the follow-up period. Conclusions Monthly intravitreal injections of ranibizumab may improve BCVA and macular edema. There was no adverse event during the follow-up duration.
Objective To study the therapeutic effect of Roux-en-Y gastric bypass （RYGB） on type 2 diabetes mellitus （T2DM） rats and explore the possible mechanism of vaspin in RYGB on T2DM. Methods Twenty SD rats with T2DM and 20 age- and sex-matched normal SD rats were randomly divided into 4 groups according to the random digits table：T2DM-RYGB group， T2DM-sham operation （SO） group，RYGB group，and SO group，10 rats in each group. Fasting plasma glucose （FPG） level，serum insulin （INS） level，vaspin level，and homeostasis model of insulin resistance （HOMA-IR） were determined before operation and on week 4，8 after operation，respectively．At the same time，the correlation between vaspin and the indicators （FPG，INS，or HOMA-IR） was analyzed．Results Compared the indicators after operation with before operation，the FPG level，INS level，vaspin level，and HOMA-IR were not significantly different between the T2DM-RYGB group and T2DM-SO group （P＞0.05） or between the RYGB group and SO group （P＞0.05），but the FPG level，INS level，vaspin level，and HOMA-IR in the T2DM-RYGB group and T2DM-SO group were significantly higher than those in the RYGB group （P＜0.05） and SO group （P＜0.05），respectively. On week 4 after operation，the FPG level，INS level，vaspin level，and HOMA-IR decreased in the T2DM-RYGB group，except for the FPG level，the other indexes had no significant differences as compared with the values before operation. On week 8 after operation，the FPG level，INS level，vaspin level，and HOMA-IR further decreased in the T2DM-RYGB group，there were significant differences of these indicators between before operation and on week 8 after operation. Compared the indicators after operation with before operation，the FPG level，INS level，vaspin level，and HOMA-IR were not statistically significant （P＞0.05） in the T2DM-SO group，RYGB group，or SO group. The changes in serum vaspin level correlated positively with those in INS and HOMA-IR before operaion and on week 4，8 after operaion in the T2DM-RYGB group and T2DM SO group rats （P＜0.05），respectively. Conclusions RYGB surgery has a therapeutic effect on T2DM rats，and serum vaspin level decreases and insulin resistance is improved after RYGB surgery，which may be one of the mechanisms of the treatment for T2DM.
Since November 1996, 20 cases of craniofacial deformities, either from congenital or traumatic, were treated with MEDPOR surgical implant made from a linear high density polythylene. The animal experiment had shown that the MEDPOR had good organotrophic characteristics allowing tissue ingrowth. The biocompatibility studies in vitro and in vivo had shown that the MEDPOR biomaterial was free from any observable systemic or cytotoxic effect. In the clinical application, it was found that the MEDPOR could be easily modeled and maintained. Because of the ability to induce tissuee ingrowth, the tenacity and stability of the material were enhanced. A total of seven cases of cranial defects, 8 cases of periobital defects or depressed periobital regions, 2 cases of traumatic auricular defects, 2 cases of traumatic saddle nose and 1 case of maldevelopment of mandible angle were treated. All of the cases were followed up for 6 months, the results were satisfactory.
The mechanisms behind diabetic retinopathy (DR) can be ascribed primarily to retinal microvascular abnormalities, excessive inflammatory response and neurodegeneration. Circular RNA (circRNA) is a type of endogenous non-coding RNA with a special circular structure, which is mainly composed of precursor RNA after shearing and processing. It is widely present in the retina and participates in the occurrence and development of various fundus diseases. CircRNAs express in an abnormal way in retina, serving as “the sponge” for miRNA so as to play roles in dysfunction of retinal vascular, inflammatory response and neurodegeneration in the development of DR. Further studies for circRNAs in DR will illustrate pathophysiology of DR more deeply, shedding light on circRNAs becoming novel biomarkers and molecular targets for diagnosis and treatment, thus achieving the goal of early diagnosis and precise therapy of DR.
OBJECTIVE To study the immunosuppressive effect of combined therapy with FK506 and RS-61443 in rat limb allotransplantation. METHODS: A total of 101 male SD rats were randomly divided into seven groups and used as recipients, and 101 Wistar rats were used as donors. All SD rats were performed limb allotransplantation without using immunosuppressants in control group. In experimental groups (Groups 1-6), the recipients were immunosuppressed with various dosages of FK506, RS-61443 or FK506 + RS61443, after transplantation for 5 weeks. To evaluate the results, we observed circulation of the transplanted limb, the mean rejection time, the histologic grading of skin rejection of limb grafts and the survival time of limb grafts. RESULTS: The control group showed rejection signs (edema and erythema of the skin) after a mean time of 3.36 +/- 1.15 days, and the mean survival time of the allografts was only 7.00 +/- 0.78 days. In the groups only using FK506 or RS-61443, the survival time were prolonged to varying degrees, but rejection occurred even in the period of using drug. As dosage increased, the rejection could not be prevented and the damage to liver and kidney could be induced. In the group using FK506 in combination with RS-61443, only skin and muscle of limb allografts showed slight rejection sign, function of liver and kidney was not obviously affected, the mean survival time of limb allografts was prolonged to 58.76 +/- 6.81 days. CONCLUSIONS: A combination of FK506 and RS-61443 is a more potent immunosuppressive agent than FK506 oro RS-61443 in preventing the rejection of limb allografts, and it can obviously prolong the survival time of limb allografts.
Objective To explore the relationship between nasopharyngeal microecology and diseases in children with bronchial asthma. Methods A total of 41 children with asthma who were treated in Hainan Provincial Hospital of Traditional Chinese Medicine between November 2020 and March 2023 were retrospectively included in the study, and 26 healthy children undergoing adenoid examination in the same period were selected as the control group. Samples of nasal mucosa were collected from the anterior and medial side of inferior turbinate, and the expression of DEFB2, IL17A, TSLP, IL13, IL5 and T1R3 genes was analyzed by polymerase chain reaction. Nasal swabs were collected from the children, and the bacterial composition was analyzed by 16S ribosomal RNA gene sequencing. Results Compared with the control group, the rate of atopy cases in the asthma group increased significantly (53.7% vs. 19.2%, P<0.05). At the phylum level, compared with the control group, the phylum Chloroflexi, the phylum Patescibacteria, the phylum Tenericutes and the phylum Nitrospirae in the asthma group increased significantly (P<0.05), and the phylum Elusimicrobia decreased significantly (P<0.05). At the genus level, compared with the control group, the members of Bacillus (Fimnicutes), Ruminococcus (Fimnicutes), Rhodococcus (Actinobacteria), Acinetobacter (Proteobacteria), Moraxella (Proteobacteria) and Asaia (Proteobacteria) in the asthma group increased significantly (P<0.05), and the members of Enterococcus (Fimnicutes), Alkanindiges (Proteobacteria), Rickettsia (Proteobacteria), and Rhizobium (Proteobacteria) in the asthma group decreased significantly (P<0.05). Compared with the control group, the Shannon index of the asthma group decreased significantly (2.63±1.45 vs. 3.90±1.44; t=2.708, P=0.010). According to receiver operating characteristic curve analysis, the optimal cut-off point of Shannon index was 3.10. In all study populations, compared with children whose Shannon index was higher than the cut-off point, children whose Shannon index was lower than the cut-off point were characterized by increased expression of IL17A and T1R3 (P<0.05) and decreased expression of TSLP (P<0.05). Conclusion The composition and abundance of nasopharyngeal microbiota are significantly different between children with asthma and healthy control children.
Objective To evaluate the inhibited effects of small interfering RNA targeting Rac1 (Rac1-siRNA) on rat retinal neovascularization in retinae. Methods Retinal vein occlusion was induced by retinal photodynamic medthod in 25 Sprague-Dawley rats. Rac1-siRNA vector DNA was injected into the vitrous of one eye of those rats (gene intervention group), and empty vector DNA was injected into the fellow eye (blank control group). Rac1-siRNA vector was injected in other 25 SD rats without retinal vein occlusion (blank intervention group). Two weeks after injection, fluorescein isothiocyanate (FITC)-dextran was perfused into the hearts of all the rats, and the retinal wholemount was made to observe the neovascularization. The numbers of endothelial cells which break through the internal limiting membrane were counted after hematoxylin-eosin staining. Results A massive of neovascularization and FITC leakage were found in blank control group. Small part of neovascularization and a little FITC leakage were observed in the gene intervention group. Retinal vessels were normal in blank intervention group. Compared with blank contrast group and blank intervention group, the difference of the mean numbers of endothelial cells which broke through the internal limiting membrane in the gene intervention group was significant（t=? P=0.000??lt;0.05）. Conclusion Rac1-siRNA can inhibit retinal neovascularization induced by retinal vein occlusion in rats.
ObjectiveTo analyze the clinical and genetic characteristics of ADGRV1 gene mutation epilepsy.MethodsA retrospective collection of 26 patients with epilepsy diagnosed and related gene sequencing was performed in the Affiliated Hospital of Jining Medical College from January 2018 to December 2018. Five epilepsy patients with ADGRV1 mutations were screened out, and their clinical characteristics and gene mutation characteristics were summarized.ResultsA total of 5 epilepsy patients with ADGRV1 mutation were collected, including 1 male and 4 females, with an average age of (7±5.83) years. Three patients had a family history of epilepsy, and the father of the other two patients had a history of febrile seizures. 2 cases showed generalized tonic-clonic seizures, and 3 cases showed partial seizures followed by generalized seizures. The results of genetic testing revealed 7 mutation sites in the ADGRV1 gene, of which one missense mutation site c.2039A>G has been reported in the literature. Two of the 5 patients underwent epilepsy surgery, and they were still treated with multiple anti-epileptic drugs for a long time after the operation, and the other 3 patients were treated with anti-epileptic drugs for a long time. At present, 4 out of 5 patients had seizures still not under effective control, and 1 case did not relapse after being followed up for nearly 1 year.ConclusionThe clinical features of epilepsy caused by ADGRV1 gene mutation are early onset, mainly manifested as general tonic-clonic seizures or partial seizures secondary to generalized seizures, accompanied by disturbance of consciousness during seizures. The combined treatment of anti-epileptic drugs and postoperative anti-epileptic drugs is less effective. Genetic testing can guide genetic counseling and assisted diagnosis.
Objective To construct small hairpin RNA (shRNA) expression plasmid targeting rat opticin gene.Methods Four pairs of opticin oligonucleotides were synthesized and inserted into the plasmid vector, resulting into four plasmids: shRNA-1, shRNA-2, shRNA-3 and shRNA-4. Then the four constructed shRNA expression vectors and empty vector were transfected into rat ciliary non-pigment epithelium (NPE) cells by lipofectmaine 2000. Nontransfected NPE cells were set as control group.The expression of opticin mRNA and protein were measured by Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot respectively.Results The opticin mRNA expression of the shRNA-1,shRNA-2,shRNA-3,shRNA-4 group were decreased compared with the control group (F=10.239，P＝0.000)；the inhibitory rate were 85.7%，62.87%，54.87% and 48.77% respectively.The opticin protein expression of the shRNA-1,shRNA-2,shRNA-3,shRNA-4 group were also decreased compared with the control group (F=17.870，P=0.000)；the inhibitory rate were 78.7%，34.6%，31.1% and 16.8% respectively.Conclusions The shRNA-1 expression plasmid has most potent inhibitory effect on opticin expression in rat ciliary NPE cells.