A novel fluorescence homogeneous biosensing strategy was developed for simple, rapid and sensitive detection of melamine in milk by using the polythymine oligonucleotide T24 and SYBR Green Ⅰ. In the absence of melamine, the fluorescence of SYBR Green Ⅰ was weak. The interactions between the single strand oligonucleotide T24 and SYBR Green Ⅰ were weak. In contrast, the presence of melamine drove the formation of T-melamine-T folded structure and enabled the SYBR Green Ⅰ to intercalate into double-strand DNA, resulting in the enhancement of fluorescence intensity. The results revealed that the method allowed a sensitive, simple, and rapid assay of melamine with a linear response range from 0. 1 μmol/L to 10 μmol/L and a detection limit of 35 nmol/L.

Objective To observe erythropoietin (EPO) and its mRNA expression changes in rats cortical neurons when suffering hypoxia and investigate the endogenous EPO protective effect of hypoxia neuronal cells.Methods Cultured cortical neurons were prepared from hypoxia rats and divided into control,hypoxia 12,24,48,72 h group.Using immunohistochemistry,reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot,we observed EPO and its mRNA expression in cells,and also observe the culture medium containing lactate dehydrogenase to evaluate the activity of neurons in the whole process.Results Immunohistochemistry,RT-PCR and Western blot analysis showed:the EPO (20.79 ± 2.98) and its mRNA (0.78 ± 0.05) at 12 h had a basic expression in hypoxia group,compared with the control group (EPO:17.12 ± 1.99; mRNA:0.39 ± 0.05),and the difference was statistically significant (t =2.51,P ＜ 0.05 ; t =13.51,P ＜ 0.01) ; the strongest expression was observed at 48 h (EPO:28.88 ± 3.41,mRNA:1.45 ± 0.07),the difference was statistically significant (t =7.29,P ＜ 0.01 ; t =33.24,P ＜ 0.01) ; and neuronal activity was strongest.Lactate dehydrogenase activity was significantly decreased after hypoxia 72 h,and also a statistically significant difference was found between the groups at each point.Conclusions The EPO and EPO mRNA expression are increased after hypoxia in neuron cells,and may enhance the activity of neurons.Our study suggests that EPO might be involved in the development process of neuronal hypoxia and play an important role in neuronal hypoxia process.

Background Corneal biomechanical properties is important in the safety assessment of corneal refractive surgery.Corvis is a new device for measuring corneal biomechanics properties.Objective This study was to observe the correlation among corneal thickness, Corvis intraocular pressure and corneal biomechanical properties with Corvis.Methods A prospective observational study was performed.One hundred and fifty eyes of 75 patients with corneal thickness from 501 μm to 590 μm were divided into three groups according to the corneal thickness:low corneal thickness group (corneal thickness range from 501 μm to 530 μm), middle corneal thickness group (corneal thickness range from 531 μm to 560 μm) ,and high corneal thickness group (corneal thickness range from 561 μm to 590 μm);and 50 eyes of 25 patients for each group.The difference of intraocular pressure, corneal thickness and deformation amplitude (DA) among the three groups were analyzed by one-way ANOVA and the correlation among the groups were analyzed by liner regression.Results The DA in the low corneal thickness group and middle corneal thickness group were significantly higher than that in the high corneal thickness group (P ＜ 0.05).The intraocular pressure was statistically different among the 3 groups (F =9.98, P＜0.05).DA was negatively correlated with intraocular pressure and corneal thickness (r=-0.84,-0.33;both at P＜0.01), with the linear regression DA =1.69-0.04×IOP (F=366.19, t=-19.14,P＜0.01).Conclusions Corneal thickness cannot simply represent the corneal biomechanical properties in the safety assessment of corneal refractive surgery,IOP should be considered.

Objective:To understand the recognition status of cancer pain in basic medical staff from small towns to provide the basis for the improvement of cancer pain management in these areas. Methods:The medical staff of Hengjiang Town and subordinate villages was selected. The study area is situated in southwest China. Centralized questionnaires regarding cancer pain were collected and analyzed. A program and education of cancer pain were provided for these medical workers. Results: The medical staff from Hengjiang asserted that only 17%of cancer pain patients receive treatments. Approximately 70%of the medical staff did not consider the popularization and explanation of cancer pain treatment in their patients. Approximately 64%of the medical staff was not familiar with standardized cancer pain control, 87%did not believe that narcotics could suffice the need of patients, and 44%did not participate in the training for cancer pain control. Conclusion: The medical staff in Hengjiang possesses less knowledge on the importance of cancer pain. Hence, further training is necessary. The specific management of cancer pain as a part of community chronic diseases is mandatory.

Objective To explore the relationship of leptin,leptin receptor and esophageal squamous cell carcinoma,and provide a scientific basis for prevention and treatment of esophageal cancer.Methods Samples were collected from 32 patients with esophageal squamous cell carcinoma,20 healthy control subjects in Shanxi Tumor Hospital.ELISA and immunohistochemistry were used to analyze leptin and leptin receptor,respectively.The correlation between their expression and clinicopathologieal parameters was also analysized.Results Patients with esophageal squamous cell carcinoma had significandy(t =4.64,P ＜ 0.001)higher leptin levels [(4.7 ±1.9)ng/ml] compared with normal [(2.54±1.01)ng/ml] oesophagus tissues.The expression rate of leptin receptor in esophageal carcinoma and normal esophagus was 81.25 ％ and 75.00 ％ respectively,not differ significantly.The expression levels of leptin was associated with position,(l)ymphatic metastasis.Conclusion Higher leptin levels seem to represent an additional,independent risk factor for esophageal squamous cell carcinoma,leptin receptor expression on oesophageal epithelial cells provides a pathway for leptin-mediated signal transduction.

OBJECTIVETo construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.

METHODSThe total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.

RESULTSThe libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).

CONCLUSIONWe have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; Cell Surface Display Techniques ; Gene Library ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Peptide Library ; Urinary Bladder Neoplasms ; genetics ; immunology

Objective:To explore the histopathological features of ulcer plaque identified by carotid CT angiography (CTA), and to explore the relationship between ulcer plaque and ischemic stroke from the microscopic perspective.Methods:Clinical data of 124 patients with carotid artery stenosis underwent carotid endarterectomy in our hospital from September 2016 to September 2020 were collected. According to the CTA results of carotid arteries, these patients were divided into ulcer plaque group ( n=47) and non-ulcer plaque group ( n=77). The clinical data, and vulnerability index and histopathological differences of placque tissues between the two groups were compared. Results:The proportion of patients with cerebral infarction history in ulcer plaque group was significantly higher than that in non-ulcer plaque group ( P<0.05). As compared with the non-ulcer plaque group, the ulcer plaque group had significantly higher proportions of fibrous cap rupture, intra-plaque hemorrhage and large necrotic core of lipid ( P<0.05). The vulnerability index of ulcer plaque group and non-ulcer plaque group was 2.01±0.51 and 0.49±0.25, respectively, with significant difference ( t=19.300, P<0.001). Conclusion:The ulcer plaque identified by CTA is correlated with fibrous cap rupture, intra-plaque hemorrhage and large necrotic core of lipid; at the same time, ulcer plaque is unstable and these patients are trended to have ischemic stroke.

Objective To investigate the remodeling of atrial fibrosis in rheumatic heart disease (RHD) patients with atrial fibrillation and its possible mechanism.Methods The clinical data and right atrial tissue specimens were obtained from patients who had undergone mitral valve replacement for rheumatic valvular disease or who had undergone thoracotomy for congenital heart disease in the Second People's Hospital of Yibin from Oct.2013 to Oct.2015.According to the heart rhythm characteristics,the specimens obtained from these patients were divided into sinus rhythm group (RHD-SR group,18 cases),paroxysmal atrial fibrillation group (RHD-pAF group,21 cases) and persistent atrial fibrillation group (RHD-cAF group,42 cases).The right atrial tissue specimens obtained from those who had undergone thoracotomy for congenital heart disease with sinus rhythm were taken as the control group(CHD-SR group,21 cases).The structures of atrial tissues were observed under a light microscope.The picric acidsirius red staining was used to detect collagen volume fraction (CVF) of type Ⅰ and Ⅲ collagen and the type Ⅰ /Ⅲ collagen CVF ratio.The expressions of TGF-β1 protein and mRNA were detected by using Western blot and reverse transcription polymerase chain reaction (RT-PCR).Results The left atrial diameter (LAd) in the RHD-cAF group was significantly higher than that in the RHDSR group (P＜0.05),but no statistically significant difference was found in other clinical data among these groups (P＞0.05).The results of picric acid-sirius red staining demonstrated that the volume fraction of type Ⅰ and type Ⅲ1 collagen and the ratio of type Ⅰ /Ⅲ collagen were increased in CHD-SR group,RHD-SR group,RHD-pAF group and RHD-cAF group,gradually,there were statistically significant differences (P＜0.05),The expression levels of TGF-β1 protein and mRNA were increased gradually in CHD-SR,RHD-SR,RHD-pAF and RHD-cAF groups (P＜0.05).Conclusion Atrial fibrosis remodeling is an important mechanism of atrial fibrillation in patients with RHD.High expression of TGF-β1 may be involved in the remodeling of atrial fibrosis.

Objective To compare the efficacy,safety and tolerability between bromfenac sodium ophthalmic solution 0.1％ eye drop and loteprednol etabonate 0.5％ eye drop after femtosecond laser in situ keratomileusis (FS-LASIK).Methods A prospective randomized controlled trail was performed.Seventy-six patients who had been undergone FS-LASIK in Daping Hospital were enrolled from December 2013 to December 2014.They were divided into two groups randomly by using the random number table method.The bromfenac sodium group included 38 patients administered 0.1％ bromfenac sodium ophthalmic solution,and the loteprednol etabonate group comprised 38 patients treated with 0.5％ loteprednol etabonate.Patients in both groups initially administered tobramycin-dexamethasone four times a day after FS-LASIK for 1 week.Then the patients in the bromfenac sodium group were treated with bromfenac sodium twice a day until 1 month postoperatively.While the patients from the loteprednol etabonate group were treated with loteprednol etabonate twice a day until 1 month after surgery.Visual acuity,refraction,intraocular pressure and ocular manifestation were examined preoperative and 1 day,1 week,1 month,3 months,6 months after FS-LASIK.This study was approved by the medical ethics Committee of Daping Hospital.Results No significant difference was found in the rate of uncorrected visual acuity (UCVA) which achieved to preoperative best corrected visual acuity (BCVA) between the two groups at each time point after operation (x2 =8.000,P =0.613).There was no difference in spherical equivalent (SE) between the two groups postoperatively (Fgroup =1.137,P =0.288),but a significant difference was found between the different time points (Ftime =29.771,P =0.000).Postoperative SE in the two groups tended to have mild hyperopia in the early days,and then gradually tended to " zero" or mild myopia.There was a significant difference with intraocular pressure among different time points (Ftime =87.519,P =0.000),but no significant difference of intraocular pressure was found between the groups (Fgroup =1.989,P =0.161).One month after surgery,the intraocular pressure were higher than 20 mmHg in two eyes in the loteprednol etabonate group,and two eyes were 5 mmHg higher than preoperative intraocular pressure.In bromfenac group,no one was higher than 5 mmHg compared with the preoperative intraocular pressure,and the pressures of all the eyes were lower than 18 mmHg.All the patients in bromfenac group were no drug withdrawal.But three patients were forced to guide drug withdrawal due to high intraocular pressure in loteprednol group.Conclusions Bromfenac sodium is safe,effective and well tolerated after FS-LASIK.

The application of high-throughput sequencing technologies has greatly enhanced our understanding to the human microbiome. The causal relations between human microbiome and diseases have become a critical issue to elucidate disease development and develop precision medicine. Recently, the study about vaginal microbiome (the microbial flora that inhabits the female vagina) has received wide interests. It has been shown that dysbiosis of vaginal microbiome was closely related to the development of genital tract diseases. This article summarizes the interaction between vaginal microbiome and disease and the treatment for the dysbiosis of vaginal microbiome. The culturomics of virginal microbiome, engineered probiotics and synthetic microbiome were also proposed.
Female ; Humans ; Microbiota ; Probiotics ; Vagina