Objective Based on real-time PCR technique and ring primers,to establish a simple,accurate,cost-effective and easily standardized quantitative assay for quantification of HER2 mRNA,and apply to provide medication guidance for clinical tumor personalized molecular targeted therapy.Methods Screening reference gene which was stable expression in breast cancer,and optimizing the PCR reaction system.Then a real-time PCR with Eva Green for quantification of the mRNA expression levels of HER2 gene was developed.The specificity,sensitivity and reproducibility of the method were evaluated 87 specimens including 55 liquid nitrogen-frozen breast cancer tissues and 32 normal tissues were detected by the real-time quantitative reverse transcription (FQ RT)-PCR and immunohistochemistry(IHC).Results The standard curve of the method indicated a good linear relationship between the Ct value and the template concentration with the correlation coefficient being 0.997.The linear range of the system was from 101 to 106 copies/μl and the lower detection limit was 101 copies/μl.It had a high sensitivity and good specificity.The inter-assay coefficients of variation of HER and RPL37A genes were (5.93 ± 0.57)％ and (5.11 ± 0.59)％,(2.49 ±0.81)％ and (2.98 ±0.97)％ respectively.The intra-assay coefficients of variation were (5.76 ±0.58)％and (7.71 ±0.61)％,(3.75 ±0.76)％ and (4.40 ±0.96)％ respectively.Using the optimized FQ RTPCR system,HER2 gene of 87 specimens was quantificated.The sensitivity of the assay was 96.36％ (53/55),the specificity was 78.13％ (25/32),the positive predictive value was 88.33％ (53/60),the negative predictive value was 92.59％ (25/27),and the total coincidence rate between FQ RT-PCR and IHC was 89.66％ (78/87).The correlation of the results between the FQ RT-PCR and IHC was good (Kappa =0.770,P ＞ 0.05).Conclusions The method can quantify the mRNA expression levels of HER2 gene rapidly and cost-effictively with high sensitivity and specificity.It can be applied to clinical molecular diagnosis with attractive prospect.

Objective To investigate if atorvastatin is effective in reducing carotid intima media thickness (CA-IMT) in normocholesterolaemic patients with transient ischemic attack (TIA). Methods Forty-two patients with TIA were recruited. Patients were randomised to receive atorvastatin(5-10 mg/d) or placebo daily. Serum concentration of cholesterol and CA-IMT were measured before randomisation and at 6-and 18-month intervals. Results CA-IMT was significantly lower in atorvastatin group than that in placebo group at 18-month (P

AIM: To observe the effectiveness, safety and ethnic differences of 0.005% atropine eye drops combined with orthokeratology in controlling adolescents' low myopia between different ethnic groups.METHODS:A total of 246 Han and Hani patients(246 eyes)with low myopia treated in our hospital from January to October 2021 were selected, with 120 patients(120 eyes)treated with 0.005% atropine eye drops combined with orthokeratology in experimental group, and 126 patients(126 eyes)treated with orthokeratology in control group. The uncorrected visual acuity, spherical equivalent(SE), axial length(AL), intraocular pressure, tear film break-up time(BUT), corneal curvature and corneal thickness of the two groups before and 1 a after wearing lenses were observed, and the incidence of complications were recorded.RESULTS:At 1 a after wearing lenses, the changes of AL and SE in the experimental group(0.16±0.35 mm, -0.39±0.47 D)were lower than those in the control group(0.22±0.89 mm, -0.48±0.54 D), uncorrected visual acuity(LogMAR)was better than the control group(0.11±0.25 vs 0.14±0.19; P<0.05), there were differences in BUT, anterior chamber depth, corneal curvature and corneal thickness(P<0.05), but there were no differences in intraocular pressure of the two groups(P>0.05). In the Han and Hani groups, there were no differences in the changes of uncorrected visual acuity, AL and SE(P>0.05). During the follow-up period, no significant local or systemic adverse reactions occurred in the two groups, and there was no difference in the incidence of ocular complications between the two groups of patients(P>0.05).CONCLUSION: The 0.005% atropine eye drops combined with orthokeratology can effectively delay the progression of low myopia in adolescents without significant adverse reactions and ethnic differences.

Metabolic-associated fatty liver disease (MAFLD), which is previously known as non-alcoholic fatty liver disease (NAFLD), represents a major health concern worldwide with limited therapy. Here, we provide evidence that ferroptosis, a novel form of regulated cell death characterized by iron-driven lipid peroxidation, was comprehensively activated in liver tissues from MAFLD patients. The canonical-GPX4 (cGPX4), which is the most important negative controller of ferroptosis, is downregulated at protein but not mRNA level. Interestingly, a non-canonical GPX4 transcript-variant is induced (inducible-GPX4, iGPX4) in MAFLD condition. The high fat-fructose/sucrose diet (HFFD) and methionine/choline-deficient diet (MCD)-induced MAFLD pathologies, including hepatocellular ballooning, steatohepatitis and fibrosis, were attenuated and aggravated, respectively, in cGPX4-and iGPX4-knockin mice. cGPX4 and iGPX4 isoforms also displayed opposing effects on oxidative stress and ferroptosis in hepatocytes. Knockdown of iGPX4 by siRNA alleviated lipid stress, ferroptosis and cell injury. Mechanistically, the triggered iGPX4 interacts with cGPX4 to facilitate the transformation of cGPX4 from enzymatic-active monomer to enzymatic-inactive oligomers upon lipid stress, and thus promotes ferroptosis. Co-immunoprecipitation and nano LC-MS/MS analyses confirmed the interaction between iGPX4 and cGPX4. Our results reveal a detrimental role of non-canonical GPX4 isoform in ferroptosis, and indicate selectively targeting iGPX4 may be a promising therapeutic strategy for MAFLD.