ObjectiveThe detection of RNA single nucleotide polymorphism (SNP) is of great importance due to their association with protein expression related to various diseases and drug responses. At present, splintR ligase-assisted methods are important approaches for RNA direct detection, but its specificity will be limited when the fidelity of ligases is not ideal. The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection. MethodsIn this study, a dual-competitive-padlock-probe (DCPLP) assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation. To verify the method, we employed dual competitive padlock probe-mediated rolling circle amplification (DCPLP-RCA) to genotype the CYP2C9 gene. ResultsThe specificity was well improved through the competition and strand displacement of dual padlock probe, with an 83.26% reduction in nonspecific signal. By detecting synthetic RNA samples, the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L. Furthermore, clinical samples were applied to the method to evaluate its performance, and the genotyping results were consistent with those obtained using the qPCR method. ConclusionThis study has successfully established a highly specific direct RNA SNP detection method, and provided a novel avenue for accurate identification of various types of RNAs.

Rheumatoid arthritis (RA) is an autoimmune disease driven by antigens and mediated by T cells. Collagen II (CII) and fibrinogen (Fib) are the two main antigens in the pathogenesis of RA. The antigen produced after citrulline modification (Cit) is also one of the inducements to induce the body to produce a pathogenic anti-citrulline protein antibody (ACPA). To provide a reference for RA-related research, this study intends to establish an RA animal model by using CII, Cit-CII, Fib, and Cit-Fib antigens, emulsification with complete Freund's adjuvant and immunization with DBA/1 mice, respectively, to compare the pathological characteristics of RA models induced by different antigens from the aspects of pathology, imaging and serum biochemistry. Animal welfare and experimental process are in accordance with the regulations of the Experimental Animal Ethics Committee of the China Academy of Chinese Medical Sciences. The results showed that the CII, Cit-CII, and Cit-Fib induced mice all had symptoms such as joint redness and swelling, and toe deformation and the clinical score and incidence rate were higher than those of the normal group. The CII group had the most serious lesions, with a incidence rate of 100%, and the Cit-CII and Cit-Fib groups had mild symptoms, with a incidence rate of 25% and 37.5%, respectively; pathological and imaging examination results showed that the joints of mice in CII-induced group showed severe synovial inflammation, cartilage and bone destruction, while those in Cit-CII and Cit-Fib group showed only slight inflammatory infiltration, joint cavity stenosis and bone destruction; the results of serum antibody detection showed that CII, Cit-CII and Cit-Fib groups all produced high levels of anti-cyclic citrullinated peptide (CCP) antibodies, among which, Cit-Fib group > Cit-CII group > CII group > Fib group, and both Cit-CII and Cit-Fib groups produced high levels of citrullinated epitope-specific antibodies, while the total IgG level was the highest in CII group; serum ELISA and RT-PCR analysis of joint tissue showed that the expression of pro-inflammatory factors and bone destruction-related molecules increased most significantly in the CII-induced group, followed by Cit-Fib and Cit-CII. The above results showed that among the four different antigens, the symptoms and conditions of arthritis in RA mice induced by CII were the most serious, and IgG instead of anti-CCP antibody was its typical immunological feature, and CII could be the first choice for the model of RA mice; Cit-Fib has certain immunogenicity, can partially induce the symptoms and conditions of RA arthritis in mice, and produce high-level anti-CCP antibody and anti-Cit-Fib antibody, which is more suitable for the study of citrulline-related RA; although Cit-CII has certain immunogenicity, the incidence, and severity of RA arthritis induced by Cit-CII in mice are low.

OBJECTIVE To investigate the mechanism of Xiaoai Jiedu Recipe in promoting ferroptosis of tumor cells and inhibi-ting the growth of transplanted tumors in hepatocellular carcinoma mice by proteomics method.METHODS C57BL/6J mice were randomly divided into,model group,low-and high-dose groups of Xiaoai Jiedu Recipe,Xiaoai Jiedu Recipe combined with ferropto-sis inhibitor group,ferroptosis inhibitor group and cisplatin group.The H22 mouse transplantation tumor model was constructed and the drug administration interventions were as follows:the low-and high-dose groups of Xiaoai Jiedu Recipe were given Xiaoai Jiedu Reci-pe by gavage at the doses of 10 and 20 g·kg-1·d-1;the ferroptosis inhibitor group was given Liproxstatin-1 by intraperitoneal injec-tion at the dose of 10 mg·kg-1·d-1;the cisplatin group was given cisplatin by intraperitoneal injection at the dose of 10 mg·kg-1·d-1;the Xiaoai Jiedu Recipe combined with ferroptosis inhibitor group was given a gavage dose of 20 g·kg-1·d-1of Xiaoai Jiedu Recipe and an intraperitoneal injection of 10 mg·kg-1·d-1 of the ferroptosis inhibitor Liproxstatin-1;the model group was given an equal amount of saline by gavage.The drugs were administered for 11 d continuously.The tumors were stripped to calcu-late tumor inhibition rate.Pathological changes were observed by hematoxylin-eosin(HE).Mitochondrial structural changes were ob-served by transmission electron microscopy.Reactive oxygen species(ROS)levels were detected by flow cytometry.Serum was pre-pared and analysed by TMT peptide labelling combined with LC-MS/MS to find differential protein expression profiles,applying IPA software.Serum iron ions,glutathione(GSH)and malondialdehyde(MDA)levels were measured biochemically.Protein expression levels of nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HMOX1),cystine/glutamate reverse transporter protein(SLC7A11)and glutathione peroxidase 4(GPX4)were measured by protein Western blot.RESULTS The tumor growth was inhibi-ted in the low-and high-dose groups and the cisplatin group,with inhibition rates of 36.12%,51.63%and 57.43%,respectively,while the tumor growth was promoted in the ferroptosis inhibitor group,with an inhibition rate of-45.56%,and the tumor size was re-duced in the Xiaoai Jiedu Recipe combined with ferroptosis inhibitor group compared with the ferroptosis inhibitor group,with an inhi-bition rate of 18.11%.HE staining showed that apoptotic cells and a large number of vacuoles accumulated in the tumor tissues of the high-dose group and the cisplatin group.Transmission electron microscopy showed that the mitochondrial atrophy and membrane densi-ty increased to a certain extent in the low and high-dose groups of Xiaoai Jiedu Recipe.Flow cytometry results showed that ROS levels were significantly increased after the intervention of Xiaoai Jiedu Recipe(P<0.01).Proteomic tests showed that there were 129 differ-ential proteins,including 62 down-regulated proteins and 67 up-regulated proteins,in the high-dose group of Xiaoai Jiedu Recipe compared with the model group,and these differential expressions were involved in lipid metabolism,et al.The results of biochemical tests showed that the intervention of Xiaoai Jiedu Recipe increased the serum iron and MDA levels,and decreased the GSH level in mice.Western blot results showed that the protein expression levels of Nrf2 and HMOX1 increased,and those of SLC7A11 and GPX4 decreased after the intervention of the high-dose group of Xiaoai Jiedu Recipe(P<0.01).CONCLUSION Xiaoai Jiedu Recipe pro-motes ferroptosis in hepatocellular carcinoma cells by inducing the Nrf2/HMOX1 signaling pathway,regulating oxidative stress and ele-vating the level of lipid peroxidation,which may be one of its mechanisms of action in inhibiting the growth of transplanted tumors.

AIM: To evaluate the effect of 0.02% mitomycin-C(MMC)on the corneal density after transepithelial photorefractive keratectomy(Trans-PRK). METHODS: Retrospective case analysis. Selected 28 patients with 56 eyes in moderate myopia who underwent Trans-PRK surgery from January 2021 to June 2021 in our hospital. They were divided into MMC group in 28 eyes with a combination of 0.02% MMC 20s during the surgery and the control group in 28 eyes was not use MMC during the surgery. The Pentacam anterior segment analyzer was used to measured the corneal density in different diameter ranges and different thickness layers before and after surgery at 14d, and after surgery at 1 and 3mo.RESULTS: The total corneal density value of MMC group was 16.60(15.70,17.10 )before the surgery, after the surgery at 14d was 16.63(15.90,17.50 ), at 1mo was 16.57(15.10,16.70 ), at 3mo was 16.04(14.60,16.60 ). The total corneal density value of control group was 16.30(15.50,17.30 )before the surgery, after the surgery at 14d was 16.20(15.20,17.10 ), at 1mo was 16.08(14.90,16.40 )and at 3mo was 15.60(14.60,16.40 ). In the zone of 0-2mm diameter was centered on the corneal vertex, the corneal density of the two groups at 14d after the surgery was higher than those before surgery(P<0.001 ). In the zone of 2-6mm diameter, the corneal density of the two groups at 1mo and 3mo after surgery was higher than those before the surgery(P<0.001). In the zone of 6-10mm, the corneal density of the two groups at 14d, 1 and 3mo after surgery was higher than those before the surgery(P<0.001). In the layer of anterior 120 μm, the corneal density of the two groups at 1mo and 3mo after the surgery was decreased than that before surgery(P<0.01). In the middle layer, the corneal density of the two groups at 1mo after the surgery was decreased than those before surgery(P<0.01).CONCLUSION:The use of 0.02% MMC during the operation can reduce the corneal density and increase the corneal light transmittance in the early postoperative period. The occurrence and prognosis of haze can be effectively quantified by observing the changes of corneal optical density in different ranges in different time periods after operation.

Objective: To compare the short-term and long-term outcomes between robotic-assisted and laparoscopic-assisted radical right hemicolectomy in patients with adenocarcinoma of the right colon. Methods: Retrospective review of a prospectively collected database identified 288 right colon cancer patients who underwent either robotic-assisted (n=57) or laparoscopic-assisted right hemicolectomy (n=231) between October 2014 and October 2020 at Department of Gastrointestinal Surgery, the Affiliated Hospital of Qingdao University. There were 161 males and 127 females, aging (60.3±12.8) years (range: 17 to 86 years). After propensity score matching as 1∶4 between robotic-assisted and laparoscopic-assisted right hemicolectomy, there were 56 cases in robotic group and 176 cases in laparoscipic group. Perioperative outcomes and overall survival were compared between the two groups using t test, Wilcoxon rank sum test, χ² test, Fisher exact test, Kaplan-Meier method and Log-rank test, respectively. Results: The total operative time was similar between the robotic and laparoscopic group ((206.9±60.7) minutes vs. (219.9±56.3) minutes, t=-1.477, P=0.141). Intraoperative bleeding was less in the robotic group (50 (20) ml vs. 50 (50) ml, Z=-4.591, P<0.01), while the number of lymph nodes retrieved was significantly higher (36.0±10.0 vs. 29.0±10.1, t=4.491, P<0.01). Patients in robotic group experienced significantly shorter hospital stay, shorter time to first flatus, and defecation (t: -2.888, -2.946, -2.328, all P<0.05). Moreover, the overall peri-operative complication rate was similar between robotic and laparoscopic group (17.9% vs. 22.7%, χ²=0.596,P=0.465). The 3-year overall survival were 92.9% and 87.9% respectively and the 3-year disease-free survival rates were 83.1% and 82.6% with no statistical significance between the robotic and laparoscopic group (P>0.05). Conclusions: Compared to laparoscopic-assisted right hemicolectomy, robot-assisted right hemicolectomy could improve some short-term clinical outcomes. The two procedures are both achieving comparable survival.
Colectomy ; Colonic Neoplasms/surgery* ; Female ; Humans ; Laparoscopy ; Male ; Prognosis ; Propensity Score ; Retrospective Studies ; Robotic Surgical Procedures ; Treatment Outcome

Objective: To investigate the association between blood pressure and all-cause mortality in Shanxi, China. Methods: The '2002 China Nutrition and Health Survey' baseline data in Shanxi province was used. A retrospective investigation was performed in 2015. The effects of SBP and DBP on the all-cause mortality were analyzed using the Cox regression model. The hazard ratio ( Results: The follow-up rate was 76.52% over 13 years, while the cumulative mortality rate for all participants was 917.12/100,000 person-years. The mortality rose with an increasing SBP ( Conclusion Adults with SBP > 160 mmHg and DBP > 100 mmHg had a higher mortality risk. Sex and age difference was noted in both DBP and mortality risk.
Adolescent ; Adult ; Aged ; Blood Pressure ; China ; Cohort Studies ; Female ; Health Surveys ; Humans ; Hypertension/mortality* ; Male ; Middle Aged ; Mortality/trends* ; Proportional Hazards Models ; Young Adult

Objective:To observe the effect of Liuwei Dihuangtang on the mitochondrial apoptotic pathway in rats with chronic renal failure. Method:The 45 male SD rats were randomly divided into normal group （n=10）， sham group （n=10） and operation group （n=25）. The operation group received 5/6 nephrectomy， and the rats in operation group were randomly divided into the model group and the treatment group after successful modeling. The treatment group received Liuwei Dihuangtang by gavage administration， while the rest of the three groups received equal volume of distilled water by gavage administration， with a treatment course of 8 weeks. The quantitative levels of serum creatinine （SCr）， blood urea nitrogen （BUN） and urine protein in each group were determined. Hematoxylin eosin （HE） staining and Masson staining were used to observe the histopathological changes of rat kidney. The structural changes of mitochondria in renal tubular epithelial cells were observed under electron microscope. The apoptotic rate was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling（TUNEL）. The expression of B cell lymphoma -2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cytochrome C （Cyt C） and cysteine aspartic acid protease -3 （Caspase-3） were analyzed by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:As compared with the normal group， the quantitative levels of SCr， BUN and urinary protein in the model group were significantly increased （P<0.01）， and the above indicators were improved in the treatment group as compared with the model group （P<0.05）. No significant pathological changes and damage of mitochondrial structure were observed in the kidney tissues of the control group and the sham group. In the model group， there were different degrees of renal tubular atrophy and dilatation， interstitial inflammatory cell infiltration and collagen deposition， and mitochondria swelling and breaking， but such lesions were significantly alleviated in the treatment group. As compared with the normal group， apoptotic rate was increased significantly in model group （P<0.01）， the expression of Bax and Caspase-3 was significantly increased （P<0.01）， and the expression of Bcl-2 was significantly decreased （P<0.01）. As compared with the model group， apoptotic rate was significantly reduced in the treatment group （P<0.01）， the expression of Bax and Caspase-3 was significantly decreased （P<0.01）， and the expression of Bcl-2 was significantly increased （P<0.01）. In the normal group and sham group， Cyt C was mainly located in mitochondria. The expression of Cyt C in the cytoplasm of the model group was increased significantly （P<0.01）， while that of the treatment group was decreased （P<0.01）. Conclusion:Liuwei Dihuangtang can alleviate renal fibrosis and delay the progression of renal function. The mechanism of reducing apoptosis may be associated with protecting mitochondrial structure and regulating the expression of proteins related to mitochondrial apoptotic pathway.