Objective To develop an ultra-performance liquid chromatography-mass spectrometry(UPLC-MS/MS)method for the simultaneous quantification of dolutegravir,raltegravir,efavirenz,lamivudine and tenofovir in human plasma and to apply it to the therapeutic monitoring.Methods Dolutegravir-D5,raltegravir-D4,efavirenz-D5,lamivudine-13 C-15 N2 and tenofovir-D7 were used as internal standard,respectively.All samples were extracted using the protein precipitation method with acetonitrile and then diluted for analysis.Chromatographic separation was performed on Shim-pack XR-ODS Ⅲ(2.0 mmx50 mm,1.6 μm)column.Mobile phases A and B consisted of 0.1％formic acid in water and acetonitrile respectively.A programmed mobile phase gradient was used at a flow rate of 0.3 mL·min-1 and column temperature of 40 ℃.The tandem mass spectrometer was equipped with an electrospray ionization(ESI)source operating in multiple reaction monitoring(MRM)modes.After methodological validation,it can be used for therapeutic drug monitoring in HIV patients.Results There was good linearity in the validated concentration ranges of 62.5-3 000 ng·mL-1 for dolutegravir,10-500 ng·mL-1 for raltegravir,125-6 000 ng·mL-1for efavirenz,10-500 ng·mL-1 for lamivudine and 10-500 ng·mL-1 for tenofovir with the linear correlation coeffificients of determination(R2)of all higher than 0.998.The accuracy of both intra-day and inter-day studies ranged from 94.0％-109.3％,and the relative standard deviations were less than 7％.The IS-normalized matrix factor and extraction recoveries of all analytes were 95.7％-106.0％and 98.7％-104.5％at all concentrations.All analytes were stable in plasma at a certain storage environment.The trough blood concentrations of dolutegravir,efavirenz,lamivudine and tenofovir were 107.7-2 366.0,740.0-3 410.0,38.5-1 229.3,31.6-224.4ng·mL-1 in HIV patients,respectively.Conclusion The method is highly aceurate,easy to perform,low-cost,and suitable for therapeutic drug monitoring of dolutegravir,raltegravir,efavirenz,lamivudine and tenofovir in HIV patients.

Objective:To observe the lipid metabolism characteristics of type 2 diabetes mellitus (T2DM) patients with different levels of blood glucose control and preliminarily analyze their relationship with diabetic retinopathy (DR).Methods:A retrospective clinical study. From January 2019 to January 2024, 232 T2DM patients who underwent fundus examination in Department of Ophthalmology of Yichang Central People’s Hospital were included in the study. Based on the glycated hemoglobin A1c (HbA1c) test results, patients were divided into blood glucose standard group and blood glucose non standard group, with 100 and 132 cases respectively. Based on the results of fundus fluorescein angiography, patients were divided into non DR (NDR) group and DR group, with 89 and 143 cases, respectively. 100 healthy individuals who underwent physical examinations during the same period were selected as the control group. The thickness of peripapillary retinal nerve fiber layer (pRNFL) around the optic disc, the blood flow density of radial peripapillary capillaries (RPC) around the optic disc, and the thickness of ganglion cell complex (GCC) in the upper and lower parts of the optic disc and macular area were measured by optical coherence tomography angiography instrument. Fully automated biochemical analyzer was used to detect serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and HbA1c. After adjusting for confounding factors, multiple linear regression model was used to analyze the correlation between HbA1c and blood lipids. Multiple logistic regression analysis was conducted to investigate the correlation between TG, HDL-C, and the occurrence of DR.Results:Compared with the control group, both the blood glucose standard group and the blood glucose non standard group had higher levels of HbA1c ( F=8.115), TC ( F=4.373), TG ( F=20.220), and LDL-C ( F=12.271), and lower levels of HDL-C ( F=6.349), with statistically significant differences ( P<0.05). Compared with the blood glucose standard group, patients in the blood glucose non standard group had higher levels of serum HbA1c ( t=3.531), TC ( t=2.561), TG ( t=6.418), LDL-C ( t=7.880), and lower levels of HDL-C ( t=5.152), with statistically significant differences ( P<0.05). Correlation analysis showed that HbA1c was positively correlated with TC, TG, and LDL-C ( P<0.05), and negatively correlated with HDL-C ( P<0.05). Multiple logistic regression analysis showed that TG, LDL-C, and HDL-C were independent risk factors for the occurrence of DR ( Ptrend<0.05). Compared with the NDR group, the DR group had thinner GCC and pRNFL thickness in the upper part of the optic disc, and lower overall and RPC blood flow density in the upper part of the optic disc, with statistically significant differences ( t=4.964, 2.406, 2.685, 2.404; P<0.05). Correlation analysis results showed that TG, LDL-C, HDL-C, HbA1c were correlated with GCC thickness, pRNFL thickness, and RPC blood flow density ( P<0.05). Conclusions:The higher the blood glucose level in T2DM patients, the more likely they are to experience dyslipidemia. TG, LDL-C, and HDL-C are independent risk factors for the occurrence of DR. Abnormal blood lipids and blood glucose levels in T2DM patients can affect retinal nerves, blood vessels, and function.

Purpose: Oligometastatic non–small cell lung cancer (NSCLC) patients have been increasingly regarded as a distinct group that could benefit from local treatment to achieve a better clinical outcome. However, current definitions of oligometastasis are solely numerical, which are imprecise because of ignoring the biological heterogeneity caused by genomic characteristics. Our study aimed to profile the molecular alterations of oligometastatic NSCLC and elucidate its potential difference from polymetastasis. Materials and Methods: We performed next-generation sequencing to analyze tumors and paired peripheral blood from 77 oligometastatic and 21 polymetastatic NSCLC patients to reveal their genomic characteristics and assess the genetic heterogeneity. Results: We found ERBB2, ALK, MLL4, PIK3CB, and TOP2A were mutated at a significantly lower frequency in oligometastasis compared with polymetastasis. EGFR and KEAP1 alterations were mutually exclusive in oligometastatic group. More importantly, oligometastasis has a unique significant enrichment of apoptosis signaling pathway. In contrast to polymetastasis, a highly enriched COSMIC signature 4 and a special mutational process, COSMIC signature 14, were observed in the oligometastatic cohort. According to OncoKB database, 74.03% of oligometastatic NSCLC patients harbored at least one actionable alteration. The median tumor mutation burden of oligometastasis was 5.00 mutations/Mb, which was significantly associated with smoking, DNA damage repair genes, TP53 mutation, SMARCA4 mutation, LRP1B mutation, ABL1 mutation. Conclusion Our results shall help redefine oligometastasis beyond simple lesion enumeration that will ultimately improve the selection of patients with real oligometastatic state and optimize personalized cancer therapy for oligometastatic NSCLC.

Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN).AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.

Objective:To investigate the effects of probiotics on intestinal flora, intestinal function, and T lymphocyte level in patients with cervical cancer after radiotherapy.Methods:A total of 92 patients with cervical cancer who underwent pelvic radiotherapy in The Second Affiliated Hospital of Zhengzhou University from September 2020 to February 2022 were included in this study. They were randomly divided into control and experimental groups ( n = 46/group). The patients in the experimental group took probiotics during radiotherapy, while the patients in the control group did not take probiotics during radiotherapy. The amount of intestinal flora, D-lactic acid, diamine oxidase, and T lymphocyte subset levels pre- and post-radiotherapy were compared between the two groups. Urinary lactulose (L) and mannitol (M) concentrations were determined in each group. Urinary excretion ratios of L to M were calculated. Results:After 10, 15, and 20 times of radiotherapy and after all radiotherapies, the amount of Escherichia coli and Enterococcus in the experimental group was significantly lower than that in the control group ( F = 128.60, 224.99, all P < 0.05). The amount of Bifidobacteria and Lactobacilli in the experimental group was significantly higher than that in the control group ( F = 2 065.46, 948.23, both P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma D-lactic acid level in the experimental group was (9.34 ± 1.63) μg/L, (9.15 ± 1.36) μg/L, (8.68 ± 1.06) μg/L, and (8.05 ± 0.82) μg/L, respectively. After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma diamine oxidase level in the experimental group was (86.34 ± 20.25) μg/L, (84.28 ± 17.45) μg/L, (80.40 ± 13.35) μg/L, and (76.85 ± 10.87) μg/L, respectively, and urinary excretion ratio of L to M in the experimental group was (1.84 ± 0.16), (1.55 ± 0.12), (1.26 ± 0.09), (0.98 ± 0.06), respectively, all of which were significantly lower than those in the control group ( F = 121.60, 31.73, 417.84, all P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, CD4 + level in the experimental group was (39.80 ± 4.90)%, (40.92 ± 5.30)%, (42.52 ± 6.14)%, (43.83 ± 6.55)%, respectively, CD4 +/CD8 + was (1.52 ± 0.25), (1.63 ± 0.22), (1.71 ± 0.39), (1.83 ± 0.22), respectively, all of which were significantly higher than those in the control group ( F = 58.69, 31.07, all P < 0.05). Conclusion:Probiotics can improve the status of intestinal flora and intestinal barrier function in patients with cervical cancer after radiotherapy, and simultaneously improve the cellular immune function of patients.

Pulmonary hypertension is a severe disease with a wide spectrum of underlying etiologies, which was characterized by increased pulmonary arterial pressure and pulmonary vascular resistance, and eventually leads to right heart dysfunction and even death with a high mortality rate. Melatonin, as a neuroendocrine hormone, is produced primarily by the pineal gland. Melatonin, a pleotropic molecule, plays a key role in the pathophysiology of various cardiovascular diseases.The effects of melatonin which can attenuate pulmonary vascular remodeling and pulmonary artery pressure have been widely concerned by researchers in recent years. This review summarized the progress of melatonin on pulmonary hypertension.

Objective:To investigate the effect of lncRNA UCA1 on the radiosensitivity of in vitro cultured glioma cell lines SHG-44, U87 and U251 by regulating the miR-873-5p expression. Methods:The survival of glioma cells SHG-44, U87 and U251 treated with different radiation intensities (0, 2, 4, 6 and 8 Gy) was detected by colony formation assay. The expression levels of UCA1 in glioma cells SHG-44, U87 and U251 were measured by qRT-PCR. The radiation-resistant glioma cells U87 and U251 were selected for subsequent study. After silencing UCA1 expression and/or over-expressing miR-873-5p, the cell survival rate was detected by colony formation assay, and the cell apoptosis rate was determined by flow cytometry. The dual luciferase reporter gene assay and qRT-PCR were employed to verify the targeting relationship between UCA1 and miR-873-5p.Results:UCA1 was up-regulated in the radiation-resistant U87 and U251 cells. Silencing UCA1 or over-expressing miR-873-5p inhibited the survival of U87 and U251 cells, and promoted the cell apoptosis induced by radiation exposure. miR-873-5p was a target gene of UCA1, and UCA1 negatively regulated the expression of miR-873-5p. The inhibition of miR-873-5p could reverse the effect of silencing UCA1 on the radiosensitivity of glioma cells. Silencing UCA1 increased the inhibitory effect of radiation on the glioma cell U251 xenografts.Conclusion:Silencing UCA1 inhibits the survival of glioma cells and promotes the cell apoptosis by up-regulating the expression of miR-873-5p, thereby increasing the radiosensitivity of glioma cells.

Objective:To explore the effect of nursing intervention based on the integration theory of health behavior change in pelvic floor muscle exercise of patients with pelvic floor dysfunction (PFD) after early cervical cancer surgery.Methods:A total of 134 patients with pelvic floor dysfunction after early cervical cancer surgery were selected and divided into test group ( n=67) and control group ( n=67) by random digits table method. The test group received three months of pelvic floor muscle exercise based on the theory of healthy behavior change. The control group received conventional pelvic floor rehabilitation care. The Pelvic Floor Distress Inventory-Short Form 20(PFDI-20), Pelvic Floor Impact Questionnaire-7(PFIQ-7), General Self-Efficacy Scale (GSES) and Social Support Rating Scale (SSRS) were evaluated before the intervention (T0), After one month of intervention (T1) and at the end of the intervention (T2). Results:The results of repeated measurement analysis of variance showed significant group/time interaction between PFDI-20 and PFIQ-7 scores ( Finteraction=29.196,13.213, both P<0.05); significant group/time interaction between GSES and SSRS scores ( Finteraction value was 25.664,70.240, both P<0.05). At the end of the intervention, the scores of PFDI-20 and PFIQ-7 in the test group were 66.54±14.04 and 73.20±7.41, which were lower than 77.47±15.91 and 78.39±8.51 in the control group over the same period. The difference was statistically significant ( t value was -4.216, -3.765, both P<0.01); the GSES and SSRS scores of the test group were 2.86±0.30 and 50.37±2.45, which were higher than the control group (2.51±0.34 and 46.42±2.82), the differences between the groups were statistically significant ( t value was 6.447, 8.662, both P<0.01). Conclusion:The nursing intervention plan based on the integration theory of health behavior change can significantly improve the self-efficacy, social support level and quality of life of patients with PFD after early cervical cancer surgery, and enhance the short-term and long-term pelvic floor rehabilitation effect of patients.

Objective:To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p.Methods:The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR. The colorectal cancer cells SW480 and SW620 were cultured in vitro, and transfected with si-NC, si-LINC00909, miR-NC, miR-584-3p mimics, si-LINC00909, and anti-miR-NC and si-LINC00909, and anti-miR-584-3p, respectively. The cells were irradiated with a dose of 4 Gy. The cell survival fraction and sensitization enhancement ratio (SER) were detected by clone formation assay. Cell proliferation was detected by MTT assay. Cell migration and invasion were assessed by Trans well chamber assay. The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay. The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice. Results:The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated ( P<0.05), whereas the expression level of miR-584-3p was significantly down-regulated ( P<0.05). After interfering with the expression of LINC00909 or miR-584-3p overexpression, the cell survival fraction score was significantly reduced ( P<0.05), the SERs were 2.017 and 1.762, and cell proliferation, migration and invasion were suppressed (all P<0.05). Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p. After interfering with the expression of LINC00909, the weight of the transplanted tumor was significantly reduced ( P<0.05), whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p ( P<0.05). Conclusion:Interfering with the expression of LINC00909 can inhibit the proliferation, migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p, thereby enhancing the cell radiosensitivity.

Objective:To observe the efficacy and safety of urokinase arterial thrombolysis in the treatment of central retinal artery occlusion (CRAO) at different time window.Methods:A retrospective study. From January 2014 to November 2019, 157 eyes (157 CRAO patients) in the Xi’an People's Hospital (Xi’an Fourth Hospital) were included in the study. There were 120 males and 37 females, with the average age of 54.87±12.12 years. The mean onset time was 65.66±67.44 h. All patients were tested with BCVA using international standard visual acuity chart, and the results were converted into logMAR visual acuity record. The arm-retinal circulation time (A-Rct) and the filling time (FT) of retinal arterial trunk-terminal filling time were measured by FFA. The mean logMAR BCVA was 2.44±0.46, the mean A-Rct and FT were 27.72±9.78 and 13.58±14.92 s respectively. According to the time window, the patients were divided into the onset 3-72 h group and the onset 73-240 h group, which were 115 patients and 42 patients respectively. There were no statistically significant difference between the 3-72 h group and the 73-240 h group in age, A-Rct and LogMR BCVA before treatment ( χ2=-0.197, -1.242, -8.990; P=0.844, 0.369, 0.369); the difference was statistically significant in FT comparison ( χ2=-3.652, P=0.000). Urokinase artery thrombolytic therapy was performed at different time window of 3-24 h, 25-72 h, 73-96 h, 97-120 h, 121-240 h after the onset of onset. Age and A-Rct of patients with different treatment time windows were compared, and the differences were not statistically significant ( χ2=6.588, 6.679; P=0.253, 0.246).In comparison of FT and logMAR BCVA, the difference was statistically significant ( χ2 =30.150, 71.378; P=0.000, 0.000). FFA was rechecked 24 hours after treatment, BCVA was rechecked 30 days after treatment. The changes of A-Rct, FT and BCVA before and after treatment were compared and analyzed. The occurrence of adverse reactions during and after treatment were observed. The two groups of measurement data were compared. The t test was used for those with normal distribution and χ2 test was used for those with non-normal distribution. Spearman correlation analysis was used to analyze the correlation between onset time and the difference of A-Rct, FT shortening time and logMAR BCVA after treatment. Results:At 24 h after CRAO treatment, A-Rct and FT of 157 cases were 19.64±6.50 and 6.48±7.36 s respectively, which were significantly shorter than those before treatment, and the differences were statistically significant ( χ2=-16.236, -14.703; P=0.000, 0.000). The logMAR BCVA at 30 d after treatment was 1.72±0.76, which was significantly higher than that before treatment. The difference was statistically significant ( χ2=-14.460, P=0.000). After CRAO urokinase arterial thrombolysis at different time window, there were statistically significant differences in A-Rct shortening time, FT shortening time, and logMAR BCVA difference ( χ2=12.408, 24.200, 104.388; P=0.030, 0.000, 0.000). There was no statistically significant difference between the 3-72 h group and the 73-240 h group ( χ2 =-1.042, P=0.297) in shortening time of A-Rct after treatment. The difference of FT shortening time was statistically significant ( χ2=-3.581, P=0.000). The difference of logMAR BCVA was statistically significant ( χ2=-9.905, P=0.000). The results of Spearman correlation analysis showed that there was no correlation between the onset time and the shortening time of A-Rct and FT after treatment ( rp=-0.040, -0.081; P=0.436, 0.115), and negative correlation with the logMAR BCVA difference ( rp=-0.486, P=0.000). One case of intracranial hemorrhage occurred after treatment, and it improved after dehydration to reduce cerebral edema, scavenging free radicals and brain protection. Conclusions:Urokinase arterial thrombolytic therapy is effective for CRAO within time window of 3-240 h, A-Rct, FT and LogMRA BCVA are all improved. However, with the prolongation of thrombolytic therapy time window, the therapeutic effect of urokinase arterial thrombolytic therapy is decreased. The therapeutic effect of Urokinase arterial thrombolytic therapy was better within 72 h.