[Observation of the consistency between intellectualized and manual-based cognitive assessment tools in the outpatient clinic].

Objective: The intellectualized versions of the Montreal Cognitive Assessment Scale (MoCA) and the Mini-mental State Examination (MMSE) (i-MoCA/i-MMSE) were developed. The validity of this system was evaluated in a clinical sample through comparing with the manual-based assessments. Methods: A total of 88 patients [aged (66.82±11.37) years, 30 males and 58 females] were enrolled in the outpatient clinic of Xuanwu Hospital of Capital Medical University with complaints of cognitive decline, from February to October 2023. All participants completed manual-based and intellectualized assessments in a randomized order, with an interval of 2 weeks to control for the practice effect. The reliability of the intellectualized version of assessments was evaluated based on the manual-based version using the Concordance correlation coefficient (CCC). The difference between the intellectualized and the manual-based assessments was tested by the Repeated ANCOVA with demographic information controlled. The accuracy of evaluation of the i-MoCA and i-MMSE was analyzed by the Receiver Operating Characteristic (ROC) analysis. Results: High concordance was observed between the intellectualized version and the manual-based assessments (CCCMoCA=0.87, CCCMMSE=0.83). Controlling for basic demographic information, there was no significant difference in the scores of the intellectualized version and the manual-based assessments (all P>0.05). The accuracy of i-MoCA in screening patients with cognitive impairment was 94.3% (sensitivity=94.6%, specificity=78.1%), while the accuracy of i-MMSE in screening patients with cognitive impairment was 94.9% (sensitivity=94.9%, specificity=77.6%). In addition, the majority of subdomains measured by the cognitive assessments exhibited high consistency across the intellectualized the manual-based versions (CCCMoCA=0.32-0.78; CCCMMSE=0.54-0.79). Conclusion: Both the i-MoCA and i-MMSE showed high consistency and diagnostic accuracy with the manual-based versions in terms of overall cognitive function and subdomains.

[Imaging study on determining the rationality of atlantoaxial fixation angle based on the ratio of line segments between anatomical markers on lateral X-ray films].

Objective: To propose a method to determine the unreasonableness of the fixed angle in posterior atlantoaxial fusion surgery based on the ratio of line segments between anatomical landmarks of the atlantoaxial joint. Methods: A cross-sectional study was conducted. According to the inclusion criteria, a screening was performed on the database of asymptomatic volunteers who had full-spine lateral X-ray films taken at the Second Affiliated Hospital of Wenzhou Medical University from May 2016 to May 2021. A total of 207 volunteers were included, comprising 98 males with an age of (40.68±13.87) years and 109 females with an age of (42.64±14.45) years. On the lateral X-ray film, a line (L) parallel to the posterior margin of the odontoid process was drawn at the posterior edge of the lower articular surface of the axis (a), intersecting the atlas at points b, c, and d. The line segments ab, bd, bc, and the C1-C2 angle were measured, and the ratios of bd/ab and bc/ab were calculated. The ability of bd/ab and bc/ab to predict the unreasonable fixed angle of the atlantoaxial joint (≥22°) was analyzed by receiver operating characteristic (ROC) curve analysis in both male and female. The areas under the ROC curves (AUC) were calculated, and the performance of the two prediction methods was compared using the Delong's test. The cutoff value for distinguishing the unreasonableness of the C1-C2 angle and the sensitivity and specificity were calculated. Results: The ROC curve analysis in the male group showed that the AUC of bc/ab for predicting the unreasonable C1-C2 angle was 0.791 (95%CI: 0.696-0.867, P<0.001), with a cutoff value of 0.449, sensitivity of 97.3%, and specificity of 70.0%. The performance was significantly better than that of bd/ab (cutoff value 1.100, AUC=0.532, 95%CI: 0.428-0.634, sensitivity 26.3%, specificity 83.3%, P<0.001). The ROC curve analysis in the female group showed that the AUC of bc/ab for predicting the unreasonable C1-C2 angle was 0.804 (95%CI: 0.745-0.852, P<0.001), with a cutoff value of 0.488, sensitivity of 90.5%, and specificity of 58.6%. The performance was significantly better than that of bd/ab (cutoff value 0.960, AUC=0.687, 95%CI: 0.624-0.748, sensitivity 90.5%, specificity 44.8%, P=0.041). Conclusions: The bc/ab value can be used as an effective indicator to predict the unreasonable C1-C2 angle in posterior atlantoaxial fusion surgery with high diagnostic accuracy. The cutoff value for males is<0.449, and for females is<0.488.

[Imaging study on the safety of axial pedicle screw placement by the position of the screw trajectory tip on the anteroposterior and lateral radiographs].

Objective: To propose a method to judge the safety of axial pedicle screw placement based on the position of the tip of the screw trajectory on the anteroposterior and lateral X-ray radiographs. Methods: The cervical CT data of 40 patients admitted to the Second Affiliated Hospital of Wenzhou Medical University from December 2020 to December 2021 were selected, including 24 males and 16 females, with a mean age of (47.6±13.2) years. Based on the three-dimensional model reconstruction of Mimics software and its function of X-ray, the transmission of the axial pedicle screw and its anteroposterior and lateral films was simulated. The position of the tip of the simulated screw trajectory was divided into 5 regions (regions Ⅰ-Ⅴ) from the inside to the outside on the anteroposterior virtual radiographs, and the upper and lower regions (regions a, b) on the lateral virtual radiographs. By adjusting the direction of the screw, the tip of the screw was located in the corresponding 10 regions (80 screws in each area) on the virtual projections of the anteroposterior and lateral virtual radiographs respectively, and its accuracy was analyzed by CT to determine whether each screw penetrated the medial wall of the pedicle or vertebral artery foramen. The anteroposterior and lateral X-rays and postoperative CT data of 34 patients who underwent axial pedicle screw placement (67 axial pedicle screws were placed in total) from January 2014 to December 2021 were collected, including 18 males and 16 females, with a mean age of (45.8±14.1) years. The position of the tip of the screw trajectory on the anteroposterior and lateral films was divided in the same way. The number of screws in the corresponding 10 positions was counted, and CT analysis was used to determine whether each screw penetrated the medial wall of the axial pedicle or the vertebral artery foreman. Results: The results of the imaging simulation screw placement study showed that the perforation rate of the vertebral artery foramen in region Ⅳ and Ⅴ was 75.0% (120/160) and 100% (160/160), respectively, while the perforation rate of the medial wall of the axial pedicle in the region Ⅰ was 85.6%(137/160). The failure rate in regions Ⅱ and Ⅲ was relatively lower, and the performance of simulated screws located in the region a was better than those in region b. The perforation rates of the medial wall in regions (a-Ⅱ) and (a-Ⅲ) was 7.5% (6/80) and 0 (0/80), respectively, and the perforation rates of the vertebral foramen was 0 (0/80) and 21.3% (17/80), respectively. The retrospective imaging study also showed a higher rate of placement failure in regions Ⅰ, Ⅳ and Ⅴ, and relatively lower in regions Ⅱ and Ⅲ. There were total of 15 screws in region a-Ⅱ and a-Ⅲ, and no destruction of the medial wall of the axial pedicle and the vertebral artery foreman occurred there. Conclusions: Regions a-Ⅱ and a-Ⅲ are the "safety areas" of the tip of the pedicle screw trajectory in the axial vertebra. By analyzing the tip of the pedicle screw trajectory on the anteroposterior and lateral radiographs, the operator can determine the reasonable trajectory of axial pedicle screw placement, prevent the injury of the cervical spinal cord and vertebral artery, and reduce the risk of operation.

[Analysis of hepatic pathological inflammation and fibrosis condition and its influencing factors in 721 patients with chronic hepatitis B with normal ALT].

Objective: To analyze the hepatic pathological inflammation and fibrosis condition in order to explore the relationship with related clinical indicators in patients with chronic hepatitis B patients with normal alanine aminotransferase (ALT). Methods: 721 cases of chronic hepatitis B with normal ALT who were initially diagnosed in the Department of Infectious Diseases of Henan Provincial People's Hospital from August 2016 to December 2019 were retrospectively collected. Liver biopsy was performed in all patients. General data of patients such as gender, age, liver function indexes, blood routine indexes, HBsAg level, HBeAg status, HBV DNA level, spleen thickness and prothrombin time were collected. Univariate and multivariate analysis methods were used to determine the influencing factors of inflammation and fibrosis degree with liver biopsy. A receiver operating characteristic curve (ROC) was used to evaluate the established multi-factor prediction model. Alpha=0.05 was considered as a standard orientation of test. Results: The average age of 721 cases with chronic hepatitis B was 36.1±9.7 years, and the male to female ratio was 1.28/1, with inflammation and fibrosis grade mainly concentrated in G1S1 (349 cases), G1S2 (132 cases), G2S2 (119 cases), and G2S1 (57 cases). Among them, there were 349 (48.4%) cases of G1S1, and 372 (51.6%) cases of G/S≥2. The main manifestations were mild to moderate inflammation and fibrosis, and only 64 (8.88%) cases had severe G/S≥3. HBsAg level (stratified with 4 log10 IU/ml as the boundary) analyzed in 721 cases were correlated with the relevant clinical indicators stratification and liver pathological inflammation and fibrosis, and the difference was statistically significant (inflammation grade, χ2=6.182, P=0.013; Fibrosis grade, χ2=36.534, P=0.001). Univariate analysis of the relevant clinical indicators that may influence the patient's liver pathological G/S ≥2 showed the patient's age, albumin, γ- glutamyltransferase (GGT), platelet, prothrombin time (PT), spleen thickness and HBsAg level were all statistically significant (P<0.05), while multivariate analysis showed that age, GGT, PT, and spleen thickness had statistical differences (P<0.05). The prediction model was established in accordance to multivariate analysis, and the area under the ROC curve was 0.642. Maximization of the sum of sensitivity and specificity as cut-off value of Logit P=0.497, the diagnostic sensitivity, specificity, and Youden's index were 60.6%, 64.5%, and 0.252, respectively. Conclusion: More than half of patients with chronic hepatitis B with normal ALT have significant inflammation and fibrosis and require timely antiviral therapy. Age, GGT, PT and spleen thickness can help comprehensively evaluate the liver inflammation and fibrosis status among patients, but the lack of accurate prediction models suggests that more effective indicators that can help predict the inflammation and fibrosis status of such patients have yet to be discovered. Therefore, liver biopsy should still be actively performed in patients with normal ALT to confirm the diagnosis and timely treatment.

[Analysis of bone mineral density and its influencing factors in 211 patients with chronic hepatitis B treated with long-term entecavir monotherapy].

Objective: To investigate the changes of bone mineral density and its related influencing factors in chronic hepatitis B patients treated with long-term entecavir monotherapy. Methods: 211 cases with chronic hepatitis B treated with entecavir monotherapy in the Department of Infectious Diseases of Henan Provincial People's Hospital from June 2018 to September 2019 were retrospectively collected. Age, gender, body mass index, number of years of medication use, presence or absence of liver cirrhosis and current bone mineral density level (using dual-energy X-ray detection, taking lumbar L1 ~ 4 and left femur as observation region) and other related data were collected. 211 cases general situation was descriptively analyzed by case-control study design. Two independent sample t-tests were used to compare the differences in serum calcium, phosphorus, and renal function levels in patients with different medication durations. Univariate logistic regression was used to screen the influencing factors of bone mineral density level. Significant variables of univariate analysis were included in multivariate logistic regression to obtain the independent influencing factors leading to the decrease of bone mineral density level. The test level was set as α = 0.05. Results: The average age of 211 cases with chronic hepatitis B was (42.36 ± 11.10) years. The average medication time use was (2.52 ± 1.94) years. The body mass index (23.95 ± 3.11), and male-to-female ratio was 2.25/1. The incidence of liver cirrhosis was 35.5%. The incidence of low bone mass in the two observation sites (lumbar spine L1~4 and left femur) was 24.6% and 29.4%, respectively. There were statistically significant differences in serum calcium, phosphorus and renal function levels among patients with different entecavir treatment duration (≥3 years and < 3 years) (P < 0.05). Univariate analysis result showed that the influencing factors of BMD were age, the number of years of medication use, gender, liver cirrhosis (L1~4 of the lumbar spine region) and age, the number of years of medication, and gender (left femoral region). The variables that entered the two models after the multivariate analysis were age (L1~4 region of lumbar spine: OR = 2.225, left femur OR = 1.660), gender (L1~4 region of lumbar spine: OR = 3.048, left femur OR = 2.496), number of years of medication use (L1~4 region of lumbar spine: OR = 1.387, left femur OR = 1.276). Conclusion: Age, gender, and the number of years of medication use are independent factors that influence the bone mineral density of patients with chronic hepatitis B treated with long-term entecavir. Low bone mass risk at the two observation sites is 2.225 and 1.66 times the normal level for every 10 years of age increase. Compared with men, the risk of low bone mass at the two observation sites is 3.048 and 2.496 times for women, and for every additional year of medication use, the risk of low bone mass at the two observation sites is 1.387 and 1.276 times the normal level. Female patients with older age and prolonged medication use are at high risk of developing bone mineral density reduction.

[Gene mutation pattern of Gilbert's syndrome combined with viral hepatitis and its relationship with the exploration of clinical data].

Objective: To study whether gene mutation pattern of Gilbert's syndrome (GS) is combined with viral hepatitis and its relationship with relevant clinical data. Methods: Clinical data of GS patients combined with viral hepatitis who was admitted to the Department of Infectious Diseases of Henan Provincial People's Hospital from August 2013 to December 2018 was retrospectively analyzed. The relationship between gene mutation pattern, general data (age, gender, etc.) and liver biochemical indexes was analyzed. The differences of the above data in patients with or without combined viral hepatitis were analyzed. The measurement data were compared by t-test. The categorical data was compared by the χ (2) test. The median and interquartile range of non-normally distributed data was used to indicate the central and discrete tendency. Results: A total of 107 GS eligible cases data were collected. The male to female ratio was 4.94:1 (89:18). The average age of onset was (36.36 ± 12.51) years. Alanine aminotransferase and total bilirubin levels were normal or slightly elevated, while aspartate aminotransferase, alkaline phosphatase, and γ-glutamyltransferase were all within the normal range. There were 49 cases in the combined viral hepatitis group (36 cases with HBV and 13 cases with HCV), and 58 cases in the GS alone group. Total bilirubin level in GS alone group was higher than the combined viral hepatitis group (z = 0.035, P < 0.05), and there were no statistically significant differences in gender, age, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma glutamyltransferase (P > 0.05). Uridine diphosphate glucuronide transferase 1A1 (UGT1A1), specifically encoded by GS was detected in all 107 cases. Mutations was mainly occurred in the upstream promoter PBREM-3263 (-3279) (86 cases) and TATA box TA insertion mutation (71 cases), and GGA-AGA Gly71Arg (57 cases) mutation in EXON1 of the coding region. All mutation forms had manifestations of homozygous and heterozygous abnormalities. The combined incidence of main mutation forms in the genetic testing data were sequenced as: A2 + B2 + C2 (17 cases, 25.23%), A1 + B1 (17 cases, 15.89%), A2 (11 cases, 10.28%), C2 (10 Cases, 9.34%), A2 + B2 (7 cases, 6.54%), A1 + B2 (7 cases, 6.54%), C1 (7 cases, 6.54%), and there was no statistically significant difference between different mutation combinations in patients with or without hepatitis (P > 0.05). The results of total data analysis showed that the total bilirubin level in the single-site mutation group was higher than the multi-site mutation group (Z=2.019, P = 0.043), and other biochemical indicators had no effect (P > 0.05) and the differences were not statistically significant. Further analysis showed that the total bilirubin level of the single-site mutation subgroup in the GS alone group was higher than the multi-site mutation subgroup (Z = 1.999, P = 0.046), and the statistical difference was similar to the combined viral hepatitis group (P > 0.05). Different mutation combinations had no effect on biochemical indexes, and had no relationship with combined viral hepatitis (P > 0.05). Conclusion: GS is common in patients with combined viral hepatitis, and there is no significant difference between the incidence of gene mutation, mutation forms, biochemical indexes, and non-hepatitis group. The increase in the number of GS mutation sites does not aggravate the deterioration of bilirubin levels due to the decrease in the content and activity of uridine diphosphate glucuronosyltransferase, and the combination of different mutation sites does not affect the changes of various biochemical indexes, and at the same time it is not related to hepatitis.

[Features of a modified WHO/UCLA AVLT performance in amnestic mild cognitive impairment and mild Alzheimer's disease].

Objective: To compare the features of a modified WHO/UCLA AVLT performance in the cognitive normal, amnestic mild cognitive impairment (aMCI) and mild Alzheimer's disease (mild AD) patients. Method: A total of 105 cases of cognitivenormal (CN), 48 aMCI and 50 mild AD patients were included between 2016 and 2018. All subjects undertook detailed neuropsychological tests and brain MRI/CT scan. Results: The total score of five learning trials in CN, aMCI and AD groups were 53.9±6.9, 34.6±8.3 and 23.7±6.2, respectively (P<0.001). The score of 20-min delay recallwere 12.5±1.6, 4.3±3.0 and 0.6±1.0, respectively (P<0.001) in three groups. The score of cued recall were 13.0±1.4, 7.0±2.4 and 2.6±2.0, respectively (P<0.001). The score of 20-min delay recall had the largest effect sizes between CN and aMCI groups (Cohen'd=3.8, 95%CI,3.3-4.4), and CN and mild AD groups (Cohen'd=8.1, 95%CI 7.1-9.1). Cued recall had the largest effect size between aMCI and mild AD groups (Cohen'd=2.04, 95%CI 1.5-2.5). The scores of learning total score, 20-min delay recall, cued recall and recognition had the strong relationships with the scores of mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA) , but obtaining moderate relationships with Boston naming test and trail making test (TMT) and weak relationships with digit span and figure copy. Age and education had no relationship with the main indices of this modified AVLT. Conclusions: The modified WHO/UCLA AVLT is still an age and education fair test to assess memory domain function. Qualitative analysis of AVLT profiles may be useful to differentiate the CN, aMCI and mild AD in Chinese sample.

The microRNA-1908 up-regulation in the peripheral blood cells impairs amyloid clearance by targeting ApoE.

OBJECTIVE: To give a new insight into the mechanism of ApoE dysregulation and microRNA-1908 in Alzheimer's disease (AD). METHODS: Plasma ApoE levels were measured in 20 AD patients and 20 healthy controls. THP-1 was maintained in RPMI1640 with 10% fetal bovine serum. Quantitative real-time polymerase chain reaction was performed to detect 13-microRNA and ApoE mRNA in cultured cell lines. Enzyme-linked immunosorbent assay was used to measure human ApoE in the plasma or culture medium of cell lines and also used to quantify the human Aß42 in the culture medium of cell lines. RESULTS: We found plasma ApoE level reduced in AD patients (2.28 vs 3.78 µg/mL, P < .001), and microRNA-1908 was up-regulated in AD patients and was negatively associated with plasma ApoE (r = -0.32, P = .012). In human macrophage cell line THP-1 and astrocytoma cell line U87, microRNA-1908 could inhibit the mRNA and protein levels of ApoE by targeting its 3'untranslated region. Consistently, microRNA-1908 inhibits the ApoE-mediated Aß clearance. CONCLUSIONS: Our study provides new insight into the mechanism of ApoE dysregulation in AD patients, and microRNA-1908 might be a therapeutic target for AD treatment.

[A real-world study of paritaprevir/ritonavir-ombitasvir combined with dasabuvir in the treatment of genotype 1b chronic hepatitis C].

Objective: To recognize the efficacy and safety of paritaprevir/ritonavir-ombitasvir combined with dasabuvir (OBV/PTV/RTV+DSV) in the treatment of genotype 1b chronic hepatitis C. Methods: Patients with genotype 1b chronic hepatitis C who were admitted to the People's Hospital of Henan Province, Huashan Hospital of Shanghai and the Fifth Medical Center of the General Hospital of the People's Liberation Army of China between November 2017 to August 2018 were enlisted. All patients received OBV/PTV/RTV+DSV antiviral therapy. HCV RNA levels were measured at baseline, weeks 1, 2, 3, 4, 8, 12, and 24, then 12 weeks, and 24 weeks after completion of treatment; patients' comorbidity, concomitant medications, and clinical adverse events were recorded. Results: 108 patients were enrolled in the study, with an average age of 49.1 years, 44 patients were male (40.8%), 96.3% (104/108) were newly diagnosed, and four patients had previous treatment history, of whom three were treated with IFN and one with IFN + DAA. Ninety-eight cases completed 12 weeks treatment and 89 cases were in follow up for 12 weeks, after discontinuation of the drug. Overall, 89 cases (100%) achieved SVR12.One patient treated with PR and DAA had HCV RNA level of 869175 IU/mL at 4 weeks of treatment, which was significantly higher than the baseline HCV RNA level (301776IU/ML), and was judged as failure of treatment; and follow-up was discontinued. Of all enrolled patients, 19 (17.6%) had underlying diseases and 15 (13.9%) had combined medications. During treatment, adverse events (AE) occurred in 11 patients (10.1%). The main adverse events were pruritus and elevated bilirubin. Conclusion: Combined antiviral therapy (OBV/PTV/RTV+DSV) of 12 weeks are highly effective with good safety profile in the treatment of Chinese patients with genotype 1b chronic hepatitis C.

Effects of calcitriol on experimental spinal cord injury in rats.

STUDY DESIGN: Experimental, controlled, animal study. OBJECTIVES: To evaluate the effects of calcitriol on oxidative stress, apoptosis, autophagy and locomotor recovery in rats after spinal cord injury (SCI). SETTING: China. METHODS: Ninety female rats were randomly divided into three groups. Laminectomy only was performed in the control group. The SCI group received laminectomy as well as spinal cord compression injury. In the calcitriol group, SCI rats received an intraperitoneal injection of calcitriol (2 µg kg(-1)day(-1)). Oxidative stress was assessed by the tissue superoxide dismutase (SOD) activity and the contents of glutathione (GSH) and malondialdehyde (MDA). The extent of apoptosis was assessed by immunohistochemistry for C-caspase3, TUNEL staining and western blotting for C-caspase3, Bax and Bcl2. Transmission electron microscopy was used to examine autophagosomes in the injured spinal cord of calcitriol-treated rats. Autophagy was detected by western blotting for LC3-II, Beclin1 and p62. Histological changes were assessed by haematoxylin and eosin staining and Nissl staining. Functional recovery was reflected by the Basso, Beattie and Bresnahan locomotion rating scale and the inclined plane test. RESULTS: With calcitriol treatment, oxidative stress was decreased, SOD activity and GSH content were increased and MDA content was decreased. Moreover, apoptosis was inhibited in the SCI plus calcitriol group. However, a higher level of autophagy was detected in the lesions of the calcitriol group compared with the SCI group. Histological damage and neuron loss after SCI were reduced in calcitriol-treated rats, and functional recovery was significantly promoted in the calcitriol group compared with controls. CONCLUSIONS: Calcitriol promotes locomotor recovery after SCI by reducing oxidative stress and inhibiting apoptosis, as well as promoting autophagy.

[Profiles of cognitive and language impairment of logopenic and non-fluent variant of primary progressive aphasia].

OBJECTIVE: To decipher the cognitive and linguistic feature of logopenic variant primary progressive aphasia (lv-PPA) and nonfluent variant primary progressive aphasia (nfv-PPA) and to explore the extent to which cognitive and language impairment contribute to the dysfunction of activity of daily living(ADL). METHODS: Seven lv-PPA and five nfv-PPA were enrolled in memory clinic of Xuanwu Hospital, Capital Medical University from January 2015 to January 2016 accordig to the international consensus criteria for PPA and its three subtypes. 20 age-matched normal controls (NC) were included. Both the patients and the NC completed a battery of neuropsychological test, lingusitic test and brain magnetic resonance imaging. All the patients conducted (11)C Pittsburgh compound B (PiB) PET imaging. RESULT: Lv-PPA patients were characterized by deficits in lexical retrieval and long sentenses repetition, while nfv-PPA were with motor speech apraxia and phonetic distortion. Compared with nfv-PPA, lv-PPA patient displayed more severe cognitive deficit with younger onset of age (56±5 vs 61±5, P<0.05) , rapid decline of MMSE score within 1.5 years and pariental cortex dysfunctions such as ideomotor praxis, Gerstmann syndrome and contructional apraxia. Correlation analysis indicated that there was more significant association between pariental cortex dysfunction and ADL/mini-mental state examination(MMSE) than that of language deficit(r=-0.868, r=-0.922; r=0.312, r=-0.257). All seven lv-PPA were PiB-PET positive and five nfv-PPA were negative. CONCLUSION: This study enriched the chinical and linguistic characterization of lv-PPA and nfv-PPA, which has implication for diagnosis, disease management and treatment for clinicians.

Low adenosine triphosphate activity in CD4+ cells predicts infection in patients with lupus nephritis.

OBJECTIVES: The ImmuKnow (Cylex) assay has been reported to predict the risk of infection in some diseases; however, it is uncertain whether ImmuKnow can predict the risk of infection in lupus nephritis (LN) patients receiving immunosuppressive therapy. METHODS: The ImmuKnow Immune Cell Function Assay (Cylex, Inc., Columbia, MD, USA) was applied to measure the activity of CD4+ T cells, as a marker of global immune-competence. The correlation between changes in T cell activation and the relative risk of over-immunosuppression as well as infection was studied. The amount of adenosine triphosphate (ATP) produced by CD4+ T cells in response to phytohemagglutinin (PHA) was measured for 74 LN patients without infection, 22 LN patients with severe infection (i.e. required hospitalisation), and 28 healthy controls. RESULTS: No correlation was found between the ATP level and systemic lupus erythematosus (SLE) activity. The mean ATP level was significantly lower in LN patients with infection than that in healthy controls (p<0.01) and non-infected LN patients (p<0.01). The mean ATP level in non-infected LN patients was not significantly different compared to healthy controls. A cut-off ATP value of 300 ng/mL predicted infection in LN patients with a specificity of 77% and a sensitivity of 77%. Multi-variable partial correlation coefficient between the ATP assay and severe infection was r =-0.040, p<0.001; CRP was r=0.962, p<0.001. CONCLUSIONS: The ImmuKnow assay may be effective in identifying an increased risk of infection in LN patients but is not correlated with SLE activity. Combined CRP value will increase the diagnostic rate of severe infection in SLE. Larger studies are required to establish clinical advantages of this assay in SLE treatment.

[The correlation of CD49d expression pattern with molecular genetics and hotspot gene mutants in patients with chronic lymphocytic leukemia].

Objective: To explore the correlation of CD49d expression patterns with molecular genetics and hotspot gene mutants in patients with chronic lymphocytic leukemia. Methods: The expression of CD49d was detected by flow cytometry and grouped into homogeneous, bimodal, negative and positive expression. Panel fluorescence in situ hybridization (FISH) was used for molecular genetics analysis and next-generation sequencing (NGS) was conducted for gene mutation detection. Results: There were 43 patients (23.89% ) with positive CD49d expression, 137 patients (76.11% ) with negative CD49d expression, 96 patients (53.33% ) with homogeneous CD49d expression and 84 patients (46.67% ) with bimodal CD49d expression. Compared with patients in the CD49d negative group, patients in the CD49d positive group had higher Rai stage (P=0.048) and higher proportion of spleen enlargement (P=0.030) . Compared with patients with homogeneous expression of CD49d, patients with bimodal expression of CD49d had a higher proportion of spleen enlargement (P=0.009) . The expression rate of 11q22- in bimodal CD49d(-) group was significantly higher than that in homogeneous CD49d(-) group (24.29% vs 10.45% , P=0.043) . The incidence of +12 in homogeneous CD49d group was higher than that in bimodal CD49d group (16.67% vs 5.95% , P=0.035) . The incidence of +12 in homogeneous CD49d(+) group was higher than that in bimodal CD49d(-) group (17.24% vs 4.29% , P=0.045) . The incidence of +12 in homogeneous CD49d(-) group was higher than that in bimodal CD49d(-) group (16.42% vs 4.29% , P=0.024) . BIRC3 mutation rate in CD49d positive group was higher than that in CD49d negative group (11.63% vs 2.92% , P=0.037) . Conclusion: There were significant correlations between CD49d and 11q22-, +12 and BIRC3 gene mutation. Patients with bimodal CD49d were more correlated with poor prognosis indexes.

[Study of cytogenetics and molecular biology in typical and atypical immunophenotypic chronic lymphocytic leukemia].

Objective: To analyze the differences in immunophenotype, cytogenetics, and molecular biology between typical and atypical immunophenotype chronic lymphocytic leukemia (CLL) , and explore the correlation of cytogenetic anomalies with gene mutations. Methods: This study included 488 patients diagnosed in the First Affiliated Hospital of Nanjing Medical University between November 2014 and May 2021. Of these, 382 patients scored 4-5 points, which was typical CLL (tCLL) , and 106 scored 3 points, which was atypical CLL (aCLL) as per the Royal Marsden Hospital Immunomarker Integral System. Peripheral blood cells were collected for immunophenotype by multiparameter flow cytometry in 488 patients, fluorescence in situ hybridization (FISH) was employed to detect cytogenetic anomalies in 359 patients, and gene mutations were detected by next-generation sequencing (NGS) in 330 patients. Results: The positive rates of CD10, CD22, CD49d, CD81, and FMC7 were significantly higher in the aCLL compared with the tCLL group (P=0.020, P<0.001, P<0.001, P=0.027, and P<0.001, respectively) , while the positive rates of CD5, CD23, CD148, and CD200 were lower in the former compared to the latter (P<0.001, P=0.017, P=0.041, and P<0.001, respectively) . aCLL exhibited a higher frequency of trisomy 12 and lower frequency of del (13q14) compared to the tCLL group (P<0.001 and P<0.001, respectively) . Moreover, aCLL patients also showed a higher incidence of NOTCH1 mutations than the tCLL patients (P=0.038) , while no statistically significant differences in other gene mutations occurred between the two groups. No significant differences in overall survival (OS) and treatment-free survival (TFS) occurred between aCLL and tCLL using Kaplan-Meier analysis (P>0.05) . Conclusion: aCLL has characteristic immunophenotype, cytogenetic, and somatic mutation that differ from tCLL, and this can provide reliable information for the diagnosis and differential diagnosis between the two groups.

Differential analyses of angiogenesis and expression of growth factors in micro- and macrovascular endothelial cells of type 2 diabetic rats.

AIMS: This study observed the relationship of angiogenesis and differential expression of growth factors and their receptors in micro- and macrovascular endothelial cells of diabetic and normal rats. MAIN METHODS: Myocardial microvascular endothelial cells (MMVEC) and aortic endothelial cells (AEC) were isolated from type 2 diabetic-Goto-Kakizaki (GK) rats and age-matched normal Wistar rats. In vitro and in vivo Angiogenesis assay were used to observe the difference between GK rats and Wistar rats. mRNA and protein expression were analyzed by Real-time RT-PCR and Western blotting. KEY FINDINGS: MMVEC but not AEC of diabetic rats had reduced abilities of angiogenesis in vitro. Real-time RT-PCR showed increased mRNA levels of VEGF, fms-like tyrosine kinase (Flt-1) and kinase insert domain containing receptor (Flk-1) in GK-MMVEC, but not the hypoxia-inducible factor-1alpha (Hif-1alpha), basic fibroblast growth factor (bFGF), fibroblast growth factor receptor 1 (FGFR1), Angiopoietin-1(Ang-1), Angiopoietin-2(Ang-2), Tie-1 and Tie-2. In contrast, Western blotting showed decreased protein levels of VEGF and receptors, including the phosphorylation of receptors. No significant differences in the expression of theses genes were observed between AEC from diabetic and control rats. Anti-rat VEGF antibodies inhibited MMVEC angiogenic function including cell proliferation, adhesion, migration, scratch wound healing and capillary-like tube formation. The in vivo angiogenesis assay had similar results. SIGNIFICANCE: These result indicated that decreased expression of VEGF and its receptors caused by post-transcription disorder in MMVEC may be responsible for diabetic impaired cardiac angiogenesis.

MicroRNA-320 expression in myocardial microvascular endothelial cells and its relationship with insulin-like growth factor-1 in type 2 diabetic rats.

1. The aim of the present study was to determine the role of myocardial microvascular endothelial cells (MMVEC) in impaired angiogenesis of type 2 diabetic Goto-Kakizaki (GK) rats. 2. A microRNA (miRNA) microarray was used to assess miRNA expression in MMVEC from GK and Wistar rats. Upregulation of miRNA-320 was observed in MMVEC from GK rats using real-time reverse transcription-polymerase chain reaction (RT-PCR). 3. So far, nine miRNAs have been reported to target angiogenic factors and/or receptors, including kinase insert domain containing receptor (Flk-1), insulin-like growth factor 1 (IGF-1) and insulin-like growth factor 1 receptor (IGF-1R). The predicted genes targeted by miR-320 include Flk-1, IGF-1 and IGF-1R. Western blot analysis and RT-PCR were used to analyse the protein and mRNA expression, respectively, of the putative genes IGF-1 and IGF-1R. The expression of IGF-1 and IGF-1R proteins decreased significantly in diabetic MMVEC. However, the expression of IGF-1 mRNA increased rather than decreased. The mRNA expression of IGF-1R did not differ significantly between diabetic and control MMVEC. 4. Transfection of an miR-320 inhibitor into MMVEC from GK rats confirmed that miR-320 impaired angiogenesis. The proliferation and migration of diabetic MMVEC improved after transfection of the miR-320 inhibitor. In addition, the miR-320 inhibitor significantly increased the expression of IGF-1 protein, but had no effect on the expression of IGF-1R. 5. Eleven miRNAs were upregulated in MMVEC from GK rats compared with those in Wistar rats: let-7e, miR-129, miR-291-5p, miR-320, miR-327, mir-333, miR-363-5p, miR-370, miR-494, miR-503 and miR-664. 6. The results indicate that upregulation of miR-320 in MMVEC from GK rats may be responsible for the inconsistency between the expression of IGF-1 protein and mRNA and therefore related to impaired angiogenesis in diabetes. Transfection of an miR-320 inhibitor may be a therapeutic approach for the treatment of impaired angiogenesis in diabetes.

Human cytomegalovirus glycoprotein genotypes in the genital tract tissue of tubal pregnancy patients.

This study investigated human cytomegalovirus (HCMV) glycoprotein genotypes in the genital tract tissue of 125 tubal pregnancy patients. The HCMV glycoprotein-B N-terminus (gBn, 54 - 485 NT), gB endoprotease cleavage site (gBclv, 1284 - 1600 NT) and glycoproteinH (gH, -58 - 213 NT) gene fragments were amplified by nested polymerase chain reaction and sequenced to identify gB and gH genotypes. Of 16 gBn-positive samples, four were gBn1, one was gBn2 and 11 were the gBn3 genotype. Of 13 positive gBclv samples, seven were gBclv1, two were gBclv2 and four were the gBclv3 genotype. Of 20 positive gH samples, 10 were gH1, six were gH2 and four were a combined gH1/gH2 genotype. In 10 of the samples that were positive for the gBn and gBclv genotypes, the gBn and gBclv genotypes were not consistent (four were gBclv1-gBn3). This study showed that: (i) HCMV infection with the gB1 - gB3 glycoprotein genotypes is present in tubal pregnancy; (ii) the gBclv and gBn genotypes are not strictly consistent; and (iii) intragenetic variability within the gB gene due to homologous recombination occurs frequently.

Strategies for the in vitro evolution of protein function: enzyme evolution by random recombination of improved sequences.

Sets of genes improved by directed evolution can be recombined in vitro to produce further improvements in protein function. Recombination is particularly useful when improved sequences are available; costs of generating such sequences, however, must be weighed against the costs of further evolution by sequential random mutagenesis. Four genes encoding para-nitrobenzyl (pNB) esterase variants exhibiting enhanced activity were recombined in two cycles of high-fidelity DNA shuffling and screening. Genes encoding enzymes exhibiting further improvements in activity were analyzed in order to elucidate evolutionary processes at the DNA level and begin to provide an experimental basis for choosing in vitro evolution strategies and setting key parameters for recombination. DNA sequencing of improved variants from the two rounds of DNA shuffling confirmed important features of the recombination process: rapid fixation and accumulation of beneficial mutations from multiple parent sequences as well as removal of silent and deleterious mutations. The five to sixfold further enhancement of total activity towards the para-nitrophenyl (pNP) ester of loracarbef was obtained through recombination of mutations from several parent sequences as well as new point mutations. Computer simulations of recombination and screening illustrate the trade-offs between recombining fewer parent sequences (in order to reduce screening requirements) and lowering the potential for further evolution. Search strategies which may substantially reduce screening requirements in certain situations are described.

Chronic social defeat stress increases dopamine D2 receptor dimerization in the prefrontal cortex of adult mice.

The present study aimed to examine the effects of chronic social defeat stress on the dopamine receptors and proteins involved in post-endocytic trafficking pathways. Adult mice were divided into susceptible and unsusceptible groups after 10 days of social defeat stress. Western blot analysis was used to measure the protein expression levels of dopamine D2 receptors (D2Rs), a short (D2S) and a long form (D2L) and, D2R monomers and dimers, dopamine D1 receptors (D1Rs), neuronal calcium sensor-1 (NCS-1) and G protein-coupled receptor-associated sorting protein-1 (GASP-1), and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the mRNA expression levels of D2S, D2L, D2R monomers and dimers, and D1Rs in different brain areas. We observed increased expression of D2S, D2L and D2Rs dimers in the prefrontal cortex (PFC) of susceptible and/or unsusceptible mice compared with controls. The only significant findings with regard to mRNA expression levels were lower expression of D2S mRNA in the amygdala (AMYG) of susceptible and unsusceptible mice compared with controls. The present study demonstrated that chronic social defeat stress induced increased expression of D2S, D2L, and D2R dimers in the PFC of susceptible and/or unsusceptible mice.