AIM: To investigate the correlation between serum levels of secreted frizzled-related protein-4(SFRP-4), chitinase 3-like protein 1(CHI3L1)and diabetic retinopathy(DR)in patients with type 2 diabetes mellitus.METHODS: Prospective study. A total of 103 DR patients who admitted to our hospital from October 2018 to October 2023 were selected as DR group, including 39 cases of early DR, 42 cases of mid DR, and 22 cases of late DR; 98 patients with simple type 2 diabetes were selected as DM group, 101 healthy individuals were selected as the control group, and baseline data and clinical indicators were collected. Enzyme linked immunosorbent assay(ELISA)was applied to detect serum levels of SFRP-4 and CHI3L1.RESULTS: The levels of serum SFRP-4, CHI3L1, triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), glycated hemoglobin(HbA1C), fasting plasma glucose(FPG), homeostasis model assessment of insulin resistance(HOMA-IR)in the DR group were higher than those in the DM group and the control group(all P<0.05); the serum levels of SFRP-4, CHI3L1, TG, LDL-C, HbA1C, FPG, HOMA-IR in the DM group were higher than those of the control group(all P<0.05). The course of disease, TG, LDL-C, HbA1C, FPG, HOMA-IR, SFRP-4, and CHI3L1 of late stage of DR were higher than those in the middle and early stages of DR(all P<0.05). The serum levels of SFRP-4 and CHI3L1 in DR patients were positively correlated with disease course, TG, LDL-C, HbA1C, FPG, HOMA-IR and DR staging(all P<0.05). The area under the curve(AUC)of serum SFRP-4, CHI3L1, and combined diagnostic DR was 0.809, 0.801, and 0.898, respectively. SFRP-4 and CHI3L1 were independent risk factors of DR(P<0.05).CONCLUSION: The levels of serum SFRP-4 and CHI3L1 are closely related to DR in patients with type 2 diabetes. The higher levels of SFRP-4 and CHI3L1 suggested that patients have a higher risk of DR.

OBJECTIVE: To analyze the differences in reflux patterns in 24-hour esophageal pH-impedance monitoring in patients with non-erosive reflux disease (NERD), reflux hypersensitivity (RH) and functional heartburn (FH) and explore the possible mechanism of symptoms in patients with heartburn and negative endoscopic findings. METHODS: Seventy-nine patients with heartburn as the main symptoms but negative endoscopic findings, including 35 with NERD, 16 with RH and 28 with FH, were enrolled in this study.All the patients underwent 24-h esophageal pH-impedance monitoring and esophagogastroscopy, and the results were compared among the 3 groups. RESULTS: Acid reflux episode was significantly increased and weakly alkaline reflux episode was significantly decreased in NERD group in comparison with RH group and FH group ( CONCLUSIONS Patients with NERD, RH and FH had different reflux patterns.Acid reflux is predominant in the NERD, while weakly alkaline reflux is significantly increased RH and FH.In patients with normal esophageal acid exposure but without symptoms or without recorded symptoms during esophageal pH-impedance monitoring, analysis of the total reflux episode, mixed reflux episode, proximal acid reflux episode and percentage can help in the differential diagnosis between RH and FH.
Electric Impedance ; Esophageal pH Monitoring ; Gastroesophageal Reflux/diagnosis* ; Heartburn/etiology* ; Humans ; Hydrogen-Ion Concentration

ObjectiveTo develop a two-dimensional high-performance liquid chromatography （2D-HPLC） for simultaneous determination of the contents of four kinds of Uncaria alkaloids： rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine. MethodsThe 2D-HPLC apparatus was comprised of a first chromatographic column in version Aston SC2 （3.5 mm×25 mm， 5 μm）， an intermediate column in version Aston SH C₁₈ （3.5 mm×10 mm， 5 μm）， and an analytical column in version Aston SCB （4.6 mm×125 mm， 5 μm）. The mobile phase of the first and second liquid chromatography system were CAA-1 and mixed mobile phase （V _{BPI-1 basic mobile phase} ∶ V _{MPI-1 mobile phase} ∶ V _{OPI-1 organic mobile phase} = 45∶14∶41）. The chromatographic parameters included a flow rate of 1.0 mL/min， a column temperature of 40℃， a wavelength of 254 nm， an injection volume of 500 μL and a detection time of 9.5 min. ResultsThe linear ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 9.77~10 000.00 ng/mL （r=0.999 6）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL（r=0.999 6）， respectively. The relative standard deviation （RSD） of precision， stability and repeatability were all less than 5.00%. The accuracy was 95.20%~104.01%， and the recovery rate was 93.63%~101.38%. ConclusionThe 2D-HPLC developed for simultaneous determination of four kinds of alkaloids in Uncaria is simple and accurate， which can be used as a new method for quality control of Uncaria.

ObjectiveTo determine the contents of four kinds of indole alkaloids （rhynchophylline， isorhynchophylline， corynoxeine， isocorynoxeine） in Uncaria by high-performance liquid chromatography-mass spectrometry-automatic internal standard （HPLC-MS-AIS）. MethodsChromatographic separation was performed using C₁₈ column （3.0 mm×50 mm， 3.3 μm）， and the mobile phase， comprising 0.1% formic acid aqueous solution-acetonitrile （82∶18， V/V）， was eluted at a flow rate of 0.5 mL/min and column temperature of 30℃. Mass spectrometric detection was performed using an electrospray ionization source and positive multiple-reaction monitoring mode at a voltage capillary of 4 000 V. The mass-to-charge ratio （m/z） transition was 385.25/160.10 for rhynchophylline， 385.30/160.10 for isorhynchophylline， 383.25/160.15 for corynoxeine and 383.25/160.15 for isocorynoxeine， respectively. The injection volume was kept constant at 2 μL. ResultsThe linear concentration ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 2.30~600.00 ng/mL （r=0.999 3）， 2.30~600.00 ng/mL （r=0.999 2）， 2.47~650.00 ng/mL （r=0.999 4） and 2.47~650.00 ng/mL （r=0.999 2）， respectively. The relative standard deviation （RSD） of precision and stability were all lower than 5.00%， the accuracy ranged from 92.40% to 104.10%， and the average recovery was 95.90%~104.60%. ConclusionHPLC-MS-AIS method is simple and accurate for the determination of four kinds of alkaloids in Uncaria， and can be used as a new method for quality control of Uncaria.

BACKGROUND: Lung cancer has the highest mortality in China. Different treatments are of great significance to the prognosis of patients. By comparing stage Ia non-small cell lung cancer (NSCLC) patients' survival rates for ablation and for sub-lobectomy, we studied the difference in the effects of the two treatments on patient prognosis. METHODS: Using the Surveillance, Epidemiology, and End Results (SEER) database, we screened eligible patients with stage Ia NSCLC from January 2004 to December 2015. Then, 228 patients treated with ablation and 228 patients treated with sub-lobotomy were then selected based on propensity score matching. After stratification, matching, and adjustment the Kaplan-Meier analysis was performed to compare the overall survival rates of patients treated with the two procedures. RESULTS: The Kaplan-Meier survival analysis showed that there is a significant difference between the ablation group and the sub-lobectomy group (P<0.05). In the univarlable analysis, the hazard ratio (HR) of sub-lobotomy group was 0.571 (95%CI: 0.455-0.717) compared with the ablation group. Patients treated with sub-lobectomy had a 0.571 times greater risk of adverse outcomes than those treated with ablation. In the multivariable analysis, the HR for sub-lobectomy group was 0.605 (95%CI: 0.477-0.766) compared with the ablation group. Patients treated with sub-lobectomy had a 0.605 time greater risk of adverse outcomes than those treated with ablation. The results suggested that the overall survival rate of patients with stage Ia NSCLC treated with sub-lobotomy was higher than that of patients treated with ablation. CONCLUSIONS This study suggests that there is a significant difference in overall survival of stage Ia NSCLC patients treated with ablation and with sub-lobotomy. Patients treated with sub-lobotomy for stage Ia NSCLC had higher overall survival than those treated with ablation.

Objective: To study the expression of interleukin-2(IL-2), soluble interleukin-2 receptor (sIL-2R), determine the alteration of erythrocytic immunity and T cell subgroup in the blood of outer circulation in patients with hypertensive cerebral hemorrhage so and to probe into the relationship between them, and to explore the clinical significance. Methods: Enzyme linked immnunosorbent assay (ELISA) was used to determine the content of IL-2 and sIL-2R. The immunoadsorption was employed to examine the erythrocytic immune activity and its regulating function. Streptavidin-peroxidase(S-P) was used to determine the cell number of CD3 (cluster of differentiation3), CD4 and CD8. Results: The content of IL-2 in the group with hypertensive cerebral hemorrhage was significantly lower than that in the control group (P<0.01), and the content of sIL-2R increased. Red blood cell C3b receptor (RBC. C 3bR) and RBC immune adherence enhancing factor (RFEB) dropped greatly (P<0.01), while RBC immune complex rosette (RBC. ICR) and RBC immune adherence inhibiting factor (RFIR) increased greatly. The cell number of CD3 and CD4 decreased (P<0.01) and there was no obvious change in CD8 (P<0.05). Conclusion: The decrease of immune function was observed in patients with hypertensive cerebral hemorrhage. The determination of the content of IL-2, sIL-2R, erythrocytic immunity and the activity of T subgroup has an important clinical significance in the occurrence, development, treatment, and prognosis of hypertensive cerebral hemorrhage.

Objective: To explore the role of the new regulator pseudogene phosphatase and tensin homolog pseudogene 1 (PTENp1) in the regulation of PTEN mRNA and gene expression in oral squamous cell carcinoma (OSCC) so as to provide a new target for the prevention, treatment and outcome of OSCC. Methods: First, we collected 42 specimens of OSCC and normal tissues, extracted RNA, detected the expressions of PTENp1, PTEN and miR-21 by qRT-PCR, and studied their correlation by Pearson correlation analysis. HEK293 cells were cultured and transfected with luciferase plasmid of 3'UTR of PTEN and mimic or inhibitor of miR-21 or full-length PTENp1 3'UTR plasmid, respectively. The regulatory role of PTENp1 in PTEN-miR-21 axis and its cancer promoting function were verified by luciferase activity test. Results: qRT-PCR showed that the expressions of PTEN (85.7%) and PTENp1 (90.4%) were significantly repressed in the OSCC tissues while miR-21 expression (76.2%) was remarkably increased. Pearson correlation analysis showed that PTEN expression was negatively correlated with miR-21 expression (R=0.123 5, P<0.001) but positively correlated with PTENp1 expression (R=0.051 8, P=0.01). The results of luciferase activity showed that PTEN expression was significantly up-regulated by overexpression of PTENp1, suggesting that PTENp1 could target competitive binding miR-21 to regulate PTEN expression. Results: PTENp1, as the ceRNA of PTEN, competitively binds the miR-21, which provides a new idea for predicting the early marker and targeting therapy of OSCC in the future.

ObjectiveTo study a therapeutic reference range and laboratory alert level of amisulpride in the clinical treatment of schizophrenia. MethodsPatients who met the diagnostic criteria for schizophrenia in the International Classification of Diseases， tenth edition （ICD-10） were enrolled， and all patients received amisulpride treatment. Data including age， gender， duration of treatment， single daily dose， clinical diagnosis， amisulpride concentration， the scores of the Scale for the Assessment of Positive Symptoms （SAPS）， Scale for the Assessment of Negative Symptoms （SANS） and Positive and Negative Syndrome Scale （PANSS）， the adverse reaction rate and multitherapy were collected. The concentration of amisulpride was compared within different efficacy groups and different dosage groups， meantime， the incidence rate of adverse reactions was compared within different amisulpride concentration groups， and between monotherapy and multitherapy groups. Thereafter， the therapeutic reference range and laboratory alert level of amisulpride in the clinical treatment of schizophrenia were explored via estimating the negative and positive predictive values. ResultsThe amisulpride concentration yielded no statistical difference within different dosage groups and different efficacy groups （F=0.745， 1.343， P>0.05）. The single daily dose among patients in different efficacy groups demonstrated no significant difference （F=0.902， P>0.05）. The correlation between amisulpride treatment efficacy and its concentration denoted no statistical significance （r=0.023， P=0.744）. The clinical efficacy and adverse reaction rate showed no significant difference between monotherapy group and multitherapy group （F=2.198， 0.095， P>0.05）. The concentration of amisulpride was not linearly correlated with the adverse reaction rates ［y=100x/（78.13+x）， r=0.960］. When amisulpride concentrations ranged from 100 to 600 ng/mL， the mean reduction rate was equal to or above 42%， the effective detection rate of the reference cut-off value was equal to or above 1.485， and the incidence of adverse reactions was equal to or below 85%. When amisulpride concentrations ranged from 1400 to 1800 ng/mL， there was a decreasing trend in reduction rate （all<42%） and an increasing trend in adverse reaction rate （all>85%） as the concentration of amisulpride increased. ConclusionA reference range of 100~600 ng/mL and an alert level of 1400 ng/mL are recommended for the clinical safety of amisulpride.

OBJECTIVETo investigate the effects of sorafenib on human acute promyelocytic leukemia cell NB4 and its mechanism.

METHODSThe human acute promyelocytic leukemia cell NB4 was treated with different concentrations (0, 1.5, 3, 6 and 12 µmol/L) of sorafenib, the proliferation inhibitory rate of NB4 cells was assayed by MTT, the apoptosis of NB4 was determined with flow-cytomatry after treatment; after extraction of total protein, the Western blot was performed to determine the expressions of apoptosis-relatived molecules Caspase-3, Caspase-8 and MCL-1. The mRNA expressions of Caspase-3, Caspase-8 and MCL-1 were determined by RT-PCR.

RESULTSAs compared with the control group, the proliferation of NB4 significantly decreased after treatment with different concentrations of sorafenib. The sorafenib significantly induced the apopotosis of NB4 cells in time- and dose-dependent manners. Furthermore, sorafenib treatment resulted in the obvious increase of the Caspase-3 and Caspase-8 protein and mRNA expressions, and down-regulated the MCL-1 protein and mRNA expressions in NB4 cells.

CONCLUSIONSorafenib can inhibit proliferation and induce apopotosis of human acute promyelocytic leukemia cell NB4 through the expression of Caspase-3 and Caspase-8, and down-regulation of the expression of MCL-1.

Antineoplastic Agents ; Apoptosis ; Caspase 3 ; Caspase 8 ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; T-Lymphocytes, Helper-Inducer

ObjectiveTo resolve the problems related to the abnormal interpretations of real-time fluorescence polymerase chain reaction （PCR） results for tri-allele， to formulate the interpretation methods of real-time fluorescence PCR by referring to multiplex PCR fragment analysis， so as to obtain an accurate， simple and cheap detection method for ABCB1 tri-allele. MethodsA total of 2 794 DNA samples were collected from Xi'an Mental Health Center from March 2020 to March 2021， and 5% of which were selected as experiments. Real-time fluorescence PCR method and multiplex PCR fragment analysis method were performed respectively. According to the comparison of Ct values of PCR curves and the comparison of base peak intensity in multiplex PCR fragment analysis， comparison and analysis were conducted on the interpretation results of the two methods， and samples with different interpretation results were verified， thereafter， PCR interpretation method was formulated. ResultsA total of 139 samples were collected， of which 120 alleles and 19 tri-allele were detected. The results of allele detection by two methods were absolutely consistent. In combination with the results of multiplex PCR fragment analysis， a method for the interpretation of real-time fluorescence PCR for 19 cases of tri-allele was developed as follows： when ∣∣Ct._G－Ct._T∣－∣Ct._G－Ct._A∣∣ in amplification curve was less than 3， the interpretation results were the combination of the base pairs with small Ct values； when ∣∣Ct._G－Ct._T∣－∣Ct._G－Ct._A∣∣ was greater than or equal to 3， the interpretation results were homozygotes from the base pairs with minimum Ct values. According to the interpretation method， the results of real-time fluorescence PCR were revised， and it showed 1 case of G/G， 1 case of A/A， 4 cases of T/G， 5 cases of T/A and 8 cases of T/T， which were consistent with the results of multiplex PCR fragment analysis. ConclusionReferring to the multiplex PCR fragment analysis method， the interpretation of ABCB1 tri-allele by real-time fluorescence PCR is developed， and the two interpretation methods are in good agreement.