Effect of miR1290 on the growth of endometrial cancer and the related mechanism / 中华肿瘤杂志

Objective: To explore the expression of miR1290 in endometrial cancer tissues and its relationship with the pathological grade, and to find out the effect of miR1290 on biological characteristics of endometrial cancer cells and its mechanism. Methods: A total of 38 cases of endometrioid adenocarcinoma tissues, 10 cases of adjacent tissues and 23 cases of normal endometrial tissues were collected in Provincial Hospital Affiliated to Shandong University from May 2020 to October 2020. The expression of miR1290 was detected by reverse transcription polymerase chain reaction (RT-PCR). The expressions of miR1290 in endometrial cancer cells including KLE and Ishikawa were knocked down by lentiviral transfection. Cell counting kit 8 (CCK-8) test and colony formation test were used to detect cell proliferation ability, wound healing and Transwell test were used to detect cell invasion and migration ability, western blot was used to detect the expressions of epithelial-mesenchymal transition (EMT), phospholipids acylinositide 3-kinase (PI3K)/Akt and Wnt/β-catenin pathway related proteins. Results: The relative expressions of miR1290 in endometrial cancer tissues were 5.40±3.20, which was 1.55 times of normal endometrial tissues (P<0.01) and 1.75 times of adjacent tissues (P<0.01). The relative expressions of miR1290 in 17 cases of endometrial tissues at proliferative stage and 6 cases of endometrial tissues at secretory stage were 3.00±1.08 and 4.97±0.58, respectively, and the difference was statistically significant (P<0.01). In KLE cells and Ishikawa cells, the expression of miR1290 in miR1290 knockdown (Sh-miR1290) group was decreased when compared with the negative control (Sh-NC) group. The absorbance value of Sh-miR1290 group detected by the CCK-8 method and the colony formation rate detected by the colony formation experiment were both increased, the number of cells penetrating the basement membrane in the Transwell experiment and the wound healing rate in the scratch experiment were decreased (P<0.05). In KLE cells, knockdown of miR1290 reduced the expressions of EMT-related proteins including N-cadherin, Vimentin, Snail and Slug(P<0.05), and the expressions of PI3K and P-Akt/Akt (P<0.05), while there was no significant change in the expressions of Wnt and β-catenin (P>0.05). In Ishikawa cells, knockdown of miR1290 decreased the expressions of EMT-related proteins including N-cadherin, Snail and Slug, and the expressions of Wnt and β-catenin, increased the expression of E-cadherin (P<0.05), while there was no significant change in the expressions of PI3K and P-Akt/Akt (P>0.05). Conclusions: The expressions of miR1290 in endometrial cancer tissues are higher than that in the adjacent tissues and normal endometrial tissues. Knockdown of miR1290 expression can promote the proliferation of endometrial cancer cells, but inhibit cell invasion, migration and EMT ability through the PI3K/Akt and Wnt/β-catenin pathways.

The Influence of Diabetes, Hypertension, and Hyperlipidemia on the Onset of Age-Related Macular Degeneration in North China: The Kailuan Eye Study / 生物医学与环境科学（英文）

Objective@#To analyze the prevalence of dry and wet age-related macular degeneration (AMD) in patients with diabetes, hypertension and hyperlipidemia, and to analyze the risk factors for AMD.@*Methods@#A population-based cross-sectional epidemiologic study was conducted involving 14,440 individuals. We assessed the prevalence of dry and wet AMD in diabetic and non-diabetic subjects and analyzed the risk factors for AMD.@*Results@#The prevalence of wet AMD in diabetic and non-diabetic patients was 0.3% and 0.5%, respectively, and the prevalence of dry AMD was 17% and 16.4%, respectively. The prevalence of wet AMD in healthy, hypertensive, hyperlipidemic, and hypertensive/hyperlipidemic populations was 0.5%, 0.3%, 0.2%, and 0.7%, respectively. The prevalence of dry AMD in healthy, hypertensive, hyperlipidemic, and hypertensive/hyperlipidemic populations was 16.6%, 16.2%, 15.2%, and 17.2%, respectively. Age, sex, body mass index, and use of hypoglycemic drugs or lowering blood pressure drugs were corrected in the risk factor analysis of AMD. Diabetes, diabetes/hypertension, diabetes/hyperlipidemia, and diabetes/hypertension/hyperlipidemia were analyzed. None of the factors analyzed in the current study increased the risk for the onset of AMD.@*Conclusion@#There was no significant difference in the prevalence of wet and dry AMD among diabetic and non-diabetic subjects. Similarly, there was no significant difference in the prevalence of wet and dry AMD among subjects with hypertension and hyperlipidemia. Diabetes co-existing with hypertension and hyperlipidemia were not shown to be risk factors for the onset of dry AMD.

The anti-venom potential of Andrographis paniculata (Burm.f.) Nees roots and its constituent skullcapflavone I

The anti-venom activity of Andrographis paniculata (Burm.f.) Nees roots (APR) dichloromethane crude extractsand a promising APR constituent, skullcapflavone I (SKI) was investigated by monitoring the inhibition of secretoryphospholipase A2 (sPLA2) of Naja philippinensis Taylor venom (NPV) crystallized samples. Gas chromatographymass spectrometry was used for the characterization of extracts, while molecular docking was utilized to understandanti-venom properties. Chromatographic analyses primarily revealed the presence of methoxylated flavones. NPV wasfound to have sPLA2 activity (0.0796 ± 0.0018 μmol/minutes/ml) that has been attributed to the poisonous effects.SKI (IC50: 51.1 ± 3.5 μg/ml), isolated from APR showed strong inhibitory effect on phospholipase activity comparedwith dichloromethane extracts of APR (IC50: 192.7 ± 10.9 μg/ml) indicating that SKI was the cause of the bioactivityin APR. Molecular docking simulations showed corresponding results with highly negative binding energies (−6.59 to−8.72 kcal/mol) predicted for the binding of SKI to PLA2 proteins. An important trend found was the presence of freebound Ca2+ lowered binding energies signifying that Ca2+ a has role in the binding of the SKI to PLA2 proteins. Theanti-venom property of APR and the pure compound SKI, upon further studies, could be the first line of defense in themedical protocol of snake venom neutralization.

The anti-venom potential of Andrographis paniculata (Burm.f.) Nees roots and its constituent skullcapflavone I

The anti-venom activity of Andrographis paniculata (Burm.f.) Nees roots (APR) dichloromethane crude extractsand a promising APR constituent, skullcapflavone I (SKI) was investigated by monitoring the inhibition of secretoryphospholipase A2 (sPLA2) of Naja philippinensis Taylor venom (NPV) crystallized samples. Gas chromatographymass spectrometry was used for the characterization of extracts, while molecular docking was utilized to understandanti-venom properties. Chromatographic analyses primarily revealed the presence of methoxylated flavones. NPV wasfound to have sPLA2 activity (0.0796 ± 0.0018 μmol/minutes/ml) that has been attributed to the poisonous effects.SKI (IC50: 51.1 ± 3.5 μg/ml), isolated from APR showed strong inhibitory effect on phospholipase activity comparedwith dichloromethane extracts of APR (IC50: 192.7 ± 10.9 μg/ml) indicating that SKI was the cause of the bioactivityin APR. Molecular docking simulations showed corresponding results with highly negative binding energies (−6.59 to−8.72 kcal/mol) predicted for the binding of SKI to PLA2 proteins. An important trend found was the presence of freebound Ca2+ lowered binding energies signifying that Ca2+ a has role in the binding of the SKI to PLA2 proteins. Theanti-venom property of APR and the pure compound SKI, upon further studies, could be the first line of defense in themedical protocol of snake venom neutralization.

The short-term intraocular pressure after 25G+ pars plana vitrectomy and analysis of possible influence factors in rhegmatogenous retinal detachment and proliferative diabetic retinopathy / 中华眼底病杂志

Objective To observe the short-term intraocular pressure after 25G+ pars plana vitrectomy (PPV) and analyze the possible influencing factors in rhegmatogenous retinal detachment (RRD) and proliferative diabetic retinopathy (PDR) eyes.Methods This is a retrospective case-control study.A total of 160 patients (163 eyes) of RRD and PDR who underwent 25G+ PPV were enrolled in this study.There were 88 males (89 eyes) and 72 females (74 eyes),with the mean age of(50.37± 13.24) years.There were 90 patients (92 eyes) with RRD (the RRD group) and 70 patients (74 eyes) with PDR (the PDR group).Best corrected visual acuity (BCVA) and intraocular pressure (IOP) were performed on all the patients.The BCVA was ranged from hand motion to 0.6.The average IOP was (12.61 ± 4.91) mmHg (1 mmHg=0.133 kPa).There were significant differences in crystalline state (x2=9.285,P=0.009),IOP (x2=58.45,P=0.000),history of PPV (x2=4.915,P=0.027) and hypertension (x2=24.018,P=0.000),but no significant difference in sex (x2=0.314,P=0.635) and age (x2=5.682,P=0.056) between the two groups.A non-contact tonometer has been used to measure IOP on postoperative day 1 and 3.The postoperative IOP distribution has been divided into five groups:severe ocular hypotension (≤5 mmHg),mild ocular hypotension (6-9 mmHg),normal (10-21 mmHg),mild ocular hypertension (22-29 mmHg),severe ocular hypertension (≥ 30 mmHg).Logistic regression analysis has been used to analyze the risk and protective factors.Results On the first day after surgery,there were 21 eyes (12.9％) in mild ocular hypotension,96 eyes (58.9％) in normal,22 eyes (13.4％) in mild ocular hypertension and 24 eyes (14.7％) in severe ocular hypertension.On the first day after surgery,there were 18 eyes (11.0％) in mild ocular hypotension,117 eyes (71.7％) in normal,23 eyes (14.1％) in mild ocular hypertension and 5 eyes (3.1％) in severe ocular hypertension.There was no significant difference of IOP distribution between the two groups (Z=-1.235,-1.642;P=0.217,0.101).The results of logistic regression analysis showed that silicone tamponade was a risk factor for ocular hypertension in PDR eyes on the first day after surgery [odds ratio (OR)=15.400,95％ confidence interval (CI) 3.670-64.590;P＜0.001],while intraocular lens was the risk factor for ocular hypotension in PDR eyes on third day after surgery (OR=19.000,95％CI 1.450-248.2;P=0.025).As for RRD eyes,the ocular hypotension before surgery was a risk factor for ocular hypertension on the third day after surgery (OR=3.755,95％CI 1.088-12.955;P=0.036).For all eyes,silicone tamponade (OR=0.236,95％CI 0.070-0.797),air tamponade (OR=0.214,95％CI 0.050-0.911) and inert gas tamponade (OR=0.092,95％CI 0.010-0.877) were protective factors for ocular hypotension on the first day after surgery (P=0.020,0.037,0.038);silicone tamponade was protective factor for ocular hypotension on the third day after surgery (OR=0.249,95％ CI 0.066-0.94,P=0.040);while aphakic eyes was the risk factor for ocular hypotension on third day after surgery (OR=7.765,95％ CI 1.377-43.794,P=0.020).The ocular hypotension before surgery was a risk factor for ocular hypertension on the third day after surgery (OR=4.034,95％ CI 1.475-11.033,P=0.007).Conclusions The abnormal IOP is common after 25G+ PPV with a rate from 28.3％ to 31.1％.Silicone tamponade,air tamponade and inert gases tamponade are protective factors for postoperative ocular hypotension,aphakic eye is risk factor for postoperative ocular hypotension.Ocular hypotension before surgery and silicone oil tamponade are risk factors for postoperative ocular hypertension.

Method establishment and preliminary application of sandwich ELISA for detecting Cryptosporidium parvum antigen / 中国人兽共患病学报

We established the method of Sandwich ELISA to detect Cryptosporidium parvum antigen.Purified anti-Cryp-tosporidium IgG and IgY were used as a capture antibody and detection antibody respectively to develop sandwich ELISA.A checkerboard titration study was carried out to determine the optimal conditions of ELISA.The PCR based on 18SrRNA was used to evaluate the pre-treatment effect of three methods (saturated sucrose solution floating,saturated salt water floating and PBST detergent washing).The optimum concentration of coated antibody,antigen,detection antibody and enzyme-labelled an-tibody were 1:800,2.5 μg/mL,1:100 and 1:5 000 respectively.The coating condition,antigen antibody reaction,opti-mum reaction time of enzyme-labelled antibody were 4 ℃ through the night after 37 ℃,incubated at 37 ℃ for 30 min and 45 min respectively;the optimal termination condition was 2 mol/L H2SO4,50 μL/well;TMB developed 10 minutes at room temperature.The developed sandwich ELISA has no cross reaction with the eggs/oocyts of Nematode,Coccidium and Asca-rid;coefficient of variation of intra-assay and inter-assay were all less than 10%.The results showed that the total coincidence rate of the three pre-treatment methods with nested PCR were 95.83%,91.67% and 83.33%,respectively,and the Saturated Sucrose Floatation method was the best one among the three methods,the sensitivity of the method was lower than the Cry p-tosporidium detection kit of IDEXX(6×103/mL),and whole test process was longer than the kit.While,its specificity and reproducibility were consistent with that of IDEXX kit,and the developed method was more economical.The method is simple,rapid,sensitive,and can be used for clinical epidemio-logical investigation of Cryptosporidiosis or pathogen detection.

Identification of deletion/duplication mutations in DMD gene by multiple ligation probe amplification and denaturing high-performance liquid chromatography / 中国医学科学院学报

<p><b>OBJECTIVE</b>To compare the effectiveness of using multiple ligation probe amplification (MLPA) and denaturing high-performance liquid chromatography (DHPLC) in screening the exon deletions and duplications of the DMD gene.</p><p><b>METHODS</b>MLPA technique was applied to detect exon deletions and duplications previously confirmed by denaturing high-performance liquid chromatography (DHPLC).</p><p><b>RESULTS</b>From October 2004 to October 2005, 22 unrelated DMD probands and their possible female relatives with clinical diagnosis with dystrophinopathy at our hospital entered this study. Both DHPLC and MPLA detected DMD gene depletion in 11 probands and DMD duplications in 3 probands. MLPA detected deletions and duplications in 2 probands, which were not detected by DHPLC. MLPA also successfully identified the carriage status of the potential female carriers of the probands.</p><p><b>CONCLUSION</b>Compared with DHPLC and traditional PCR techniques, MLPA is a superior tool to analyze the deletions and duplications in affected males as well as in the identification of the carriage status of potential females carriers.</p>

Sibling brother and sister both with Duchenne muscular dystrophy / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the clinical and lab features of sibling brother and sister both with Duchenne muscular dystrophy (DMD).</p><p><b>METHODS</b>We conducted comprehensive clinical and lab investigations including the test of serum enzymes, electromyography (EMG), electrocardiography, color Doppler echocardiography, HE staining of skeletal muscles, immunohistochemical study of dystrophin and utrophin, multiple ligation probe amplification (MLPA) on exon 1-79 of dystrophin gene, and short tandem repeat-poly- merase chain reaction of CA repeats located in dystrophin gene.</p><p><b>RESULTS</b>These two patients were confirmed to suffer from DMD. They were characterized by typical features of DMD including typical clinical manifestations, increased serum enzymes, EMG presenting myogenic impairment, HE staining presentation belonging to DMD, negative dystrophin in brother, and inconstantly positive on the sarcolemma of sister. Furthermore, no deletion or duplication was found in the 1-79 exons of dystrophin gene. The suffering brother and sister carried the same maternal X chromosome.</p><p><b>CONCLUSIONS</b>Carriers of DMD gene show typical clinical and laboratory manifestations of DMD. Comprehensive examinations should be performed for such carriers.</p>

Detection of genomic duplications and deletions of the DMD gene in affected males and female carriers by using mutiplex ligation-dependent probe amplification / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect genomic deletion and duplication mutations in the dystrophin gene of the Duchenne muscular dystrophy (DMD) patients and their potential female carriers.</p><p><b>METHODS</b>Genomic deletions and duplications of the DMD gene in 32 affected males and 27 potential female carriers were screened by mutiplex ligation-dependent probe amplification (MLPA).</p><p><b>RESULTS</b>Of the 32 investigated affected males, 24 were detected to have deletions of one or more exons of the DMD gene, 1 patient had a duplication from exon 5 to 55, 1 patient had a nonsense point mutation (R768X) in exon 19, the other 6 affected males were predicted to have possible disease-causing point mutations. MLPA analysis showed a DMD deletion or duplication in 18 female relatives, and the female carriers had the same deletion or duplication as their probands, respectively.</p><p><b>CONCLUSION</b>MLPA analysis is proven to be an efficient tool for identification of both affected males and female carriers of DMD rearrangements in cases in which the disease-causing mutation in the affected male was not known. It could provide useful information for the genetic counseling of the family involved.</p>

Transfection and in vitro expression of human microdystrophin gene in rat mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of human microdystrophin gene and observe its expression in rat mesenchymal stem cells (rMSCs) in vitro.</p><p><b>METHODS</b>The plasmid PBSK-MICRO containing human microdystrophin cDNA was digested by restriction endonuclease, and the resultant microdystrophin fragment was inserted into the NotI site of pcDNA3.1(+) to prepare the eukaryotic expression vector-pcDNA3.1(+)/ microdystrophin, which was identified by endonuclease digestion and sequencing. The recombinant plasmid was transfected into rMSCs via lipofectamine, and after G418 selection, the expression of microdystrophin was detected by RT-PCR and indirect immunofluorescence assay.</p><p><b>RESULTS</b>Microdystrophin gene fragment was correctly inserted into the plasmid pcDNA3.1(+), as conformed by sequencing and digestion with Not I and Hind III. The total mRNA of the transfected rMSCs was extracted and microdystrophin mRNA expression was found in the cells by RT-PCR. Indirect immunofluorescence assay for the protein expression of microdystrophin showed bright red fluorescence in the transfected rMSCs.</p><p><b>CONCLUSION</b>Eukaryotic expression plasmid pcDNA3.1(+)/microdystrophin has been constructed successfully and microdystrophin can be expressed in transfected rMSCs in vitro, which may facilitate further research of Duchenne muscular dystrophy treatment by genetically modified allogeneic stem cell transplantation.</p>

Identification of disease-causing point mutations in DMD patients' dystrophin gene without large deletions/duplications / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the disease-causing point mutations in the dystrophin gene of Duchenne muscular dystrophy (DMD) patients.</p><p><b>METHODS</b>The approach of denaturing high performance liquid chromatography (DHPLC) coupling with sequencing was used to screen the point mutations of 79 exons and the untranslated regions of dystrophin gene without large deletions/duplications, which was in 6 unrelated DMD probands from 6 DMD families.</p><p><b>RESULTS</b>Five disease-causing mutations, 697-698insGT, C616T, G1255T, C4279T, and C2302T, were ides created the new stop codons in downstream sites of mutations, respectively. In addition to the disease-causing point mutations, a point mutation T5586+61A in intron 39 was also found at patient 3, and a missense mutation A694T in exon 8 was detected at patient 5. Four point mutations, C2168+13T, 5740-13dupG, G5234A and C5280T, were also detected at patient 6 whose causative point mutation was unavailable. Seven point mutations have not been reported previously. Bi-directional PCR amplification of specific alleles (Bi-PASA) method was established to distinguish the haplotypes of heterozygote or homozygote in a single PCR reaction.</p><p><b>CONCLUSION</b>Via automated DHPLC screening or detecting the subexonic mutations in dystrophin gene is feasible to clinical laboratories, and also is a superior method in terms of sensitivity and efficiency.</p>

A survey on the diagnosis and treatment of biliary tract cancers in Shanghai / 中华外科杂志

<p><b>OBJECTIVE</b>To survey the status of diagnosis and treatment of biliary tract cancer in Shanghai.</p><p><b>METHODS</b>A clinical epidemiology investigation was carried out on 658 new cases of biliary duct cancers aged 35-74, that registered between June 1997 and May 2001 in urban Shanghai. Clinical findings were collected in 390 gallbladder cancer, 195 bile duct cancer and 73 ampullary cancer.</p><p><b>RESULTS</b>Biliary tract cancers mainly occurred in elderly patients. Ratio of males to female was 1:2.61 in gallbladder cancer, while bile duct cancer and ampullary cancer were slightly more common in men. Association with gallstones was 68.5%, 43.1% and 22.4% for gallbladder cancer, bile duct cancer and ampullary cancer, respectively. Diagnostic accuracy rate of B-ultrasonography was 63.1% in gallbladder cancer. Incidental gallbladder cancer accounted for 20%, while stage IVA and IVB patients reached up to 43.6%. Misdiagnosis rate was still high in bile duct cancer and ampullary cancer, it was 19.1% and 47.1% respectively. In addition, most patients presented jaundice at diagnosis. 69 cases (18.2%) of gallbladder cancer, 50 cases (25.6%) of bile duct cancer and 54 cases (74%) of ampullary cancer underwent radical resection, the 1-, 3- and 5-year survival rates were 58.5%, 42.8% and 40.7%, 58%, 28.3% and 11.1%, 81.5%, 39.2% and 26.9%, respectively. 79 patients with bile duct cancer underwent palliative drainage, and most cases died within 1 year. Metal endo-prostheses or plastic stents were placed into the biliary tract in 38 patients. The median survival was about 7 months.</p><p><b>CONCLUSIONS</b>It is difficult to make early diagnosis of biliary tract cancers. Standardization of the operation for gallbladder cancer must be respected. Surgical exploration should be undertaken when a bile duct cancer is suspected and there are no contraindications to surgery. Pancreatoduodenectomy should be recommended for ampullary cancer.</p>

Transcription factor Egr-1 is required for long-term fear memory and anxiety / 生理学报

The zinc finger transcription factor Egr-1 is critical for coupling extracellular signals to changes in cellular gene expression. In the hippocampus and amygdala, two major central regions for memory formation and storage, Egr-1 is up-regulated by long-term potentiation (LTP) and learning paradigms. Using Egr-1 knockout mice, we showed that Egr-1 was selectively required for late auditory fear memory while short term, trace and contextual memory were not affected. Additionally, synaptic potentiation induced by theta burst stimulation in the amygdala and auditory cortex was significantly reduced or blocked in Egr-1 knockout mice. Our study suggests that the transcription factor Egr-1 plays a selective role in late auditory fear memory.