Objective:To investigate the effects of low-frequency and high frequency repetitive transcranial magnetic stimulation (rTMS) combined with levodopa and benserazide hydrochloride on mild cognitive impairment in patients with Parkinson disease (PD).Methods:Totally 90 PD patients with mild cognitive impairment who visited from January 2020 to June 2022 were included , and they were divided into a simple drug group ( n=30), drug+ low-frequency group ( n=30), and drug+ high-frequency group ( n=30) according to the order of admission.The patients in the simple drug group were treated with oral levodopa and benserazide hydrochloride, while the patients in drug+ low-frequency and drug+ high-frequency groups were treated with low-frequency or high-frequency rTMS on the basis of oral levodopa and benserazide hydrochloride.Montreal cognitive assessment(MoCA), digital span (DS), Chinese auditory learning test (CALT), the judgment of line orientation test (JLOT) and verbal fluency test (VFT) were used to evaluate the cognitive function of patients before and after 4 weeks of treatment.SPSS 26.0 was used for statistical analysis.The paired t-test was used for intra-group comparison before and after treatment, while one-way ANOVA was used for inter-group comparison. Results:There were no significant differences in MoCA, DS anterograde, DS backward, CALT immediate recall, CALT delayed recall, JLOT, and VFT scores among patients in the simple drug group before and after 4 weeks of treatment( t=-1.157, -0.648, -0.215, -0.290, -0.154, -0.782, -0.960, all P>0.05). After 4 weeks of treatment, MoCA, DS anterograde, DS backward, CALT immediate recall, CALT delayed recall, JLOT and VFT scores in drug+ low-frequency group and drug+ high-frequency group were higher than before treatment (drug+ low frequency group: t=-16.357, -11.379, -7.999, -11.805, -16.624, -15.996, -17.241, all P<0.05; drug+ high-frequency group: t=-25.198, -13.971, -13.904, -25.831, -26.382, -20.108, -15.643, all P<0.05). There were no statistically significant differences in the scores of MoCA, DS anterograde, DS backward, CALT immediate recall, CALT delayed recall, JLOT and VFT among the three groups before treatment (all P>0.05). After treatment, there were statistically significant differences in the scores of MoCA, DS anterograde, DS backward, CALT immediate recall, CALT delayed recall, JLOT and VFT among the three groups (simple drug group : (20.37±1.96), (4.37±1.19), (2.80±0.55), (6.93±1.70), (5.17±1.09), (15.50±2.69), (10.73±1.55); drug+ low-frequency group: (23.83±2.32), (5.87±0.94), (3.87±0.73), (9.17±1.74), (8.13±1.50), (20.77±2.19), (13.30±1.73); drug+ high-frequency group: (27.17±1.64), (6.73±1.01), (4.80±0.81), (11.20±2.06), (10.03±1.54), (25.17±3.14), (15.87±2.05)) (all P<0.05). Further analysis showed that both the drug+ low-frequency and drug+ high-frequency groups had higher scores than the simple drug group, and the drug+ high-frequency group had higher scores than the drug+ low-frequency group(all P<0.05). Conclusion:The combination of drug+ low-frequency or drug+ high-frequency rTMS and drug therapy can help improve cognitive function in patients with PD, and the efficacy of drug+ high-frequency rTMS may be more significant, which provides a new therapeutic idea for clinical treatment of patients with PD.

The amount of data generated by the human experience in medical cases of classic formulas is large and the research value is high， but how to extract valuable information from these massive data is an important problem of researches on classical formulas. It is concluded that there are many problems in the current study of human experience in classical formulas， such as large amount of data， uneven quality， weak extrapolation and clinical application， which seriously affected the evidence strength of human experience in classical formulas and the contribution to experience accumulation and theoretical support. Therefore， from the structural perspective of the research questions， this paper proposed the research ideas of human experience in medical cases of classic formulas： firstly， according to the characteristics of traditional Chinese medicine （TCM） cases， based on the PICO model， which refers parti-cipants （P）， intervention （I）， comparisons （C）， and outcomes （O）， a TCM medical record evaluation tool in line with the evidence-based concept was constructed to evaluate the quality of medical cases； secondly， constructing a standardized database of human experience in classical formulas and using data mining algorithms to explore the deep relationship hidden in human experience； finally， the clinical value evaluation is carried out to verify the effectiveness of the diagnosis and treatment rules of classical formulas to achieve clinical transformation.

Objective:To elucidate the quasispecies variation of 3′ half-length genome in HIV-1 CRF103_01B-infected patients in Beijing using single genome amplification (SGA).Methods:This study enrolled six CRF103_01B-infected patients who were diagnosed during a drug resistance monitoring for newly diagnosed cases or newly treated cases with antiviral therapy in Beijing from 2017 to 2020. RNA was extracted from their plasma samples, and 3′ end of cDNA was diluted by serial dilution method after reverse transcription. Nested PCR was used to amplify the 3′ half-length genome sequences of HIV-1 quasispecies. MEGA 11 was used to construct Neighbor-Joining (NJ) tree and calculate the intrahost genetic distance. Genetic variation in HIV-1 quasispecies was visualized by online Highlighter tool. BootScan analysis was performed using Simplot 3.5 software to analyze inter-quasispecies recombination. Virus tropism was predicted by online Geno2pheno tool.Results:Among the six CRF103_01B-infected patients, five were men who have sex with men. A total of 144 3′ half-length genome SGA sequences (19-36 sequences/case) were obtained. The NJ tree based on the 3′ half-length genome of HIV-1 quasispecies revealed different degrees of genetic diversity. The HIV-1 quasispecies in BL4748-00 case of acute infection has the least variation with the intrahost distance of 0.002±0.000, showing genetic homogeneity. The quasispecies sequences from BL4981-00, BL3150-00 and BL3558-00 cases formed at least three subclusters, respectively, with different evolutionary directions, and their intrahost distance ranked from 0.031±0.004 to 0.016±0.002 (BL3150-00>BL3558-00>BL4981-00). The quasispecies sequences from the couple BL3022-00 (female) and BL3023-00 clustered into a large monophyletic cluster (bootstrap value=100%), and the intrahost distance of the latter (0.025±0.003) was higher than that of the former (0.019±0.002). Inter-quasispecies recombination was observed in BL3558-00 case. The quasispecies from the six patients were CCR5-tropic viruses.Conclusions:The diversity of quasispecies variation in CRF103_01B-infected patients is related to disease progress. Genetic homogeneity is observed in acute HIV infection, while multiple evolutionary directions are detected in chronic infection. Co-infection or superinfection cases are not found, but there are recombination events among quasispecies in some cases.

Objective:To establish the clinical laboratory genetic diagnosis procedures for Marfan syndrome (MFS) and carry out clinical laboratory genetic diagnosis for MFS families.Methods:The second generation high-throughput sequencing was used to sequence and analyze the FBN1 gene of two MFS families who visited to Fuwai Central China Cardiovascular Hospital (Heart Center of Henan People′s Hospital) from January to December 2020, and then Sanger sequencing was used to verify the second generation high-throughput sequencing results. At the same time, the sanger sequencing of mutation sites was performed on normal family members and 100 healthy people to identify the pathogenic mutations of FBN1 gene in the MFS families. The pregnant women of two families were guided for prenatal diagnosis in the second trimester of pregnancy.Results:The clinical laboratory diagnosis of MFS showed that two MFS patients had the pathogenic mutation of c.2560T>C heterozygous mutation and c.6772T>C heterozygous mutation in FBN1 gene, respectively. The mutation was not observed in 100 healthy people and normal members in two families. The prenatal diagnosis showed that there was a heterozygous mutation of FBN1 gene c.2560T>C in the first fetus of the MFS family, which was MFS. There was no mutation in the FBN1 gene in the second fetus of the MFS family, so it was recommended to continue the pregnancy. The results of postpartum follow-up were consistent with the results of clinical laboratory diagnosis.Conclusion:The clinical laboratory genetic diagnosis procedures for MFS have been established successfully, which provides an important reference for clarifying the clinical diagnosis of MFS.

Objective:To investigate the effects of femtosecond laser assistcataract extraction on the levels of inflammatory factors in aqueous humor, corneal perception and high order aberration.Methods:Forty-eight patients (56 eyes) undergoing cataract surgery in the Shouguang People′s Hospital from June 2018 to February 2020 were enrolled in this study, and they were divided into observation group (24 patients and 27 eyes, femtosecond laser cataract phacoemulsification) and control group (24 patients and 29 eyes, coaxial microincision phacoemulsification) by random number table method. The levels of prostglndin E2 (PGE2), interleukin (IL)-6 and IL-1β in aqueous humor were detected before and after operation. Upper corneal and central corneal perception thresholds were recorded before surgery, 7 d after surgery, 1 month after surgery and 3 months after surgery. High order aberrations of the whole eye and cornea at 4mm pupil diameter before and after operation were recorded.Results:After operation, the levels of PGE2, IL-6 and IL-1β in aqueous humor in the observation group were lower than those in the control group: (45.62 ± 5.71) ng/L vs. (63.65 ± 5.62) ng/L, (15.25 ± 3.21) ng/L vs. (24.59 ± 6.13) ng/L, (17.16 ± 1.28) ng/L vs. (25.74 ± 4.62) ng/L, the differences were statistically significant ( P<0.05). The upper and central corneal perception threshold of observation group after operation for 7 d were higher than those in the control group: 4.55 ± 0.50 vs. 4.20 ± 0.33, 5.68 ± 0.49 vs. 5.52 ± 0.45, the differences were statistically significant ( P<0.05). The indexes of high order aberration of the whole eye decreased in the two groups ( P<0.05), while the indexes of cornea did not change significantly. The indexes of high order aberration of the whole eye and cornea between before and after surgery between the two groups had no significant differences ( P>0.05). Conclusions:Femtosecond laser assist cataractextraction can relieve inflammation, reduce the whole eye high order aberration, but not affect the cornea, and can improve corneal perception.

Objective:To determine the role and molecular mechanism of dexmedetomidine (DEX) in postmenopausal osteoporosis (PMOP) .Methods:Twenty-seven patients with PMOP admitted to Yantai Yantaishan Hospital from Jan. 2020 to Jan. 2021 were selected as PMOP group, and 20 healthy volunteers were selected as Normal group. The differentially expressed miRNAs in PMOP were screened, clinically, the expression of miR-483-3p and catenin beta 1 (CTNNB1) in serum samples of patients with PMOP was detected by qRT-PCR. In vitro experiment, Bone marrow mesenchymal stem cells (BMSCs) were induced into osteoblasts, Dex was used to treat BMSCs and intervene the expression of miR-483-3p, CTNNB1 in BMSCs, the expression level of osteogenesis related indexes (RUNX2、OCN、OPN) was detected. After coculturing Human umbilical vein endothelial cell (HUVECs) with BMSCs, angiogenesis experiment was utilized to detect the angiogenesis ability.Results:Compared with Normal group (1±0.46) (1.03±0.44) , the expression of miR-483-3p (3.23±1.61) was increased in serum of PMOP patients while expression of CTNNB1 (0.50±0.27) was inhibited ( t=5.99, P<0.001) ( t=5.14, P<0.001) . miR-483-3p has a good diagnostic effect on PMOP (AUC=0.86, P<0.001) . After Dex treatment, miR-483-3p level was decreased in BMSCs, CTNNB1 level was increased (all P<0.05) . Dex promoted the expression of RUNX2, OCN, OPN and number of angiogenesis, but this effect was partially reversed by miR-483-3p overexpression (all P<0.05) . CTNNB1 was confirmed as a target gene of miR-483-3p, the inhibition effects of miR-483-3p overexpression on osteogenic differentiation and angiogenesis of BMSCs induced by Dex was partially reversed by CTNNB1 overexpression (all P<0.05) . Conclusion:Dex enhanced CTNNB1 level in PMOP via inhibiting miR-483-3p, subsequently promoted osteogenic differentiation and angiogenesis of BMSCs and inhibited progression of PMOP.

Objective:To investigate the prevalence and risk factors of diabetic peripheral artery disease (PAD) in patients with type 2 diabetes mellitus (T2DM) managed in primary health care in China.Methods:A total of 2 528 T2DM patients were selected using a two-stage cluster random sampling method based on the baseline survey of the "China Diabetic Foot Prevention Model Project." The study was conducted in 2015 among T2DM patients in 8 primary healthcare centers in Changshu county and Jiang'an district of Wuhan, China. Data collection methods included a questionnaire, body measurement, and blood glucose detection. The Ankle-Brachial Index (ABI) is the most widely used noninvasive vascular test. A binary logistic regression model was used to analyze the influence factors.Results:The prevalence of PAD was 11.2% among the diabetic patients managed in primary health care in the two cities. The prevalence of PAD under 55 years old, 55- years old, 65- years old, and ≥75 years old were 7.8%, 6.0%, 12.9% and 22.5%, respectively. Multivariate stepwise logistic regression identified influence factors included older age, higher education level, smoking, drinking, postprandial glucose uncontrol, and prior myocardial infarction or angina. Compared to age <55 years, the odds ratio for PAD were 0.74 for 55- years (95% CI: 0.43-1.28), 1.72 for 65- years (95% CI: 1.05-2.81), 3.56 for 75 years and above (95% CI: 2.07-6.11), respectively. Compared to patients with education in primary school and below, the odds ratio was 1.37 (95% CI: 0.97-1.94), 2.48 (95% CI: 1.73-3.55), 1.99 (95% CI: 1.26-3.13) for those with education levels of junior high school, senior high school, and college, respectively. Current smoking ( OR=1.49, 95% CI: 1.02-2.17), current drinking ( OR=0.45, 95% CI: 0.28-0.71), postprandial glucose uncontrol (2 h postprandial plasma glucose >10.0 mmol/L: OR=1.72, 95% CI: 1.22-2.43), and prior myocardial infarction or angina ( OR=2.32, 95% CI: 1.50-3.61) were influencing factors of PAD. Conclusions:Despite the high prevalence of PAD in diabetes managed in primary health care; multiple risk factors are not effectively aware of and under control. It is urgent to promote ABI screening and standardized management for diabetes, especially in primary health care.

Objective:To evaluate the effect of epidural labor analgesia administration methods on occurrence of postpartum urinary retention in nulliparous parturients through a comparison between programmed intermittent epidural bolus (PIEB) and continuous epidural infusion.Methods:Two hundred nulliparous parturients who were at full term with a singleton fetus in vertex presentation, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 22-35 yr, with body mass index of 22.4-42.6 kg/m 2, were divided into 2 groups ( n=100 each) using a random number table method: continuous epidural infusion group (group CEI) and group PIEB.Patient-controlled epidural analgesia (PCEA) was performed in active phase of labor (cervical dilatation≥1 cm) during the first stage of labor.The PCEA solution contained the mixture (10 ml) of 0.1% ropivacaine with 0.5 μg/ml sufentanil.The PCEA pump was set up to deliver a 5-ml bolus dose with a 30-min lockout interval.The analgesia solution contained the mixture (100 ml) of 0.08% ropivacaine and sufentanil 0.5 μg/ml.In group CEI, the drugs were given at 8 ml/h immediately after the initial dose.PIEB regimens were programmed as 8 ml over 80 s once an h after the initial bolus, and the administration was stopped after delivery of fetus.The labor time, consumption of analgesia solution and the incidence of postpartum urinary retention were recorded. Results:Compared with group CEI, the duration of the second stage of labor was significantly shortened, and consumption of analgesia solution and the incidence of postpartum urinary retention were decreased in group PIEB ( P<0.05). Conclusion:Compared with the continuous epidural infusion, the application of PIEB in labor analgesia can reduce the incidence of postpartum urinary retention in nulliparous parturients.

Objective:To observe the characteristics of the phagocytosis and bactericidal function of multidrug-resistant Mycobacterium tuberculosis(MDR- Mtb)-infected macrophage model, and the changes of the immune response and metabolic function in the process of phagocytosis and bactericidal function, aiming to provide reference for studying the role and mechanism of macrophages in the occurrence and development of multidrug-resistant tuberculosis(MDR-TB). Methods:We established MDR- Mtb and H37Rv-infected macrophage models, and used the colony-forming unit (CFU), Magnetic Luminex ? Assay and Cholesterol Assay kit to observe the effects on phagocytosis and bactericidal function, the secretion of Th1(IL-12/23 p40, IL-27 and TNF-α) and Th2 cytokines (IL-6 and IL-10) and cholesterol metabolism. The data were analyzed by SPSS25.0 software. The data were expressed as Mean± SD and analyzed by t test or F test. P<0.05 was considered statistically significant. Results:(1) After MDR- Mtb-infected macrophages, the intracellular CFU gradually increased and reached the highest at 24 h, while the extracellular CFU gradually decreased and reached the lowest at 24 h. The intracellular CFU at 48 h was lower than that at 24 h, while the extracellular CFU was higher than that at 24 h ( P<0.05). Both intracellular and extracellular CFU at 48 h were close to those at 4 h ( P>0.05). The intracellular CFU was lower than the H37Rv group at 8-48 h, while the extracellular CFU was higher than the H37Rv group ( P<0.05). (2) The level of IL-12/23 p40, IL-27, TNF-α, IL-6 and IL-10 of MDR-TB group were higher than those of blank group ( P<0.05), but the level of TNF-α and IL-6 at 24 h and 48 h were higher than that at 4 h ( P<0.05). IL-12/23 p40 and TNF-α at 48 h and IL-6 at 24 h were lower than those of the H37Rv group, while IL-27 at 48 h was higher than that of the H37Rv group ( P<0.05). (3) The levels of cholesterol of MDR-TB group at 24 h and 48 h were lower than those of 4 h and blank group ( P<0.05), but the level of cholesterol was similar to the H37Rv group at any time ( P>0.05). (4) TNF-α reached the highest when the intracellular CFU reached the highest at 24 h, and IL-6 reached the highest when the intracellular CFU decreased at 48 h. With the decreasing of cholesterol expression, the intracellular CFU increased and then decreased. Conclusions:MDR- Mtb could induce the phagocytosis and bactericidal function of macrophages, increase the expression of Th1 and Th2 cytokines and promote the utilization and consumption of cholesterol, but this function was weaker than that of H37Rv strain.

Objective:To study the effect of the psychological counseling on the delivery mode of single birth elderly primiparas with anxiety and depression, and to seek a new and effective intervention way to improve the vaginal delivery rate.Methods:A preliminary screening was carried out for 486 single birth elderly primiparas received in the birth cohort research center of China Medical University from April 2018 to September 2019. Among them, 274 cases had mild, moderate or severe anxiety and depression. They were randomly divided into the observation group and the control group (n=137). The control group was given routine nursing measures and simple psychological education and the intervention group was given extra psychological counseling. Psychological counseling adopted the mode of combination of online and offline. The degree of anxiety and depression and the delivery mode were compared between two groups.Results:Before intervention, the degree of anxiety and depression between the observation group and the control group showed no significant difference. After intervention, the degree of anxiety and depression in the observation group was lighter than that of the control group ( χ2 value was 12.782, 6.647, P<0.05). The proportion of cesarean section, vaginal delivery, forceps delivery accounted for 32.1% (44/137), 67.2%(92/137), 0.7%(1/137) in the observation group, and 45.3% (62/137), 54.7%(75/137), 0 in the control group. The difference between the two groups showed statistical significance ( χ2 value was 5.787, P<0.05). Conclusion:The psychological counseling for the single birth elderly primiparas with anxiety and depression can effectively improve the psychological situation and increase the vaginal delivery rate.