Objective To analyze the expression levels of serum microRNA(miR)-211 and miR-202 in patients with Alzheimer's disease and their correlation with cognitive function,anxiety and depression.Methods A total of 90 patients with Alzheimer's disease admitted to Hebei Yanda Hospital from March 2019 to March 2022 were selected as the research group.According to the Clinical Dementia Rating(CDR)score,the patients were grouped into mild group(n=24),moderate group(n=48)and severe group(n=18).Another 90 healthy individuals who underwent physical examination were collected as the control group.The expression levels of miR-211 and miR-202 in serum were compared.Pearson method and Spearman method were used to analyze serum miR-211 and miR-202 and their correlation with cognitive function,anxiety and depression.Logistic regression analysis was used to analyze the influencing factors of Alzheimer's disease.Results The expression levels of serum miR-211(0.59±0.16,1.01±0.31)and miR-202(0.35±0.10,1.00±0.32)were significantly reduced in the research group and control group,with significant differences(t=11.422,18.393,all P<0.05).Serum miR-211(0.73±0.21,0.62±0.17,0.32±0.08)expression levels,miR-202(0.51±0.15,0.33±0.10,0.19±0.04)expression levels,mini-mental state examination(MMSE)score(22.54±1.41 score,19.35±1.01 score,16.23±1.00 score)and Montreal cognitive assessment(MoCA)score(25.35±2.60 score,18.59±1.32 score,16.59±1.24 score)in the mild,moderate and severe groups gradually decreased,and the differences were statistically significant(F=32.006,46.917,163.048,163.703,all P<0.05).Compared with mild group,the serum miR-211,miR-202,MMSE and MoCA scores of severe group and moderate group were reduced,and the differences were statistically significant(t=3.685～25.375,all P<0.05).The mild,moderate and severe groups had a gradual increase in Hamilton anxiety scale(HAMA)score(12.34±1.27 score,20.59±2.09 score and 31.29±2.19 score)and Hamilton depression scale(HAMD)score(14.35±2.13 score,23.89±2.20 score and 35.35±1.21 score),and the differences were statistically significant(F=496.059,553.939,all P<0.05).According to Pearson correlation analysis,miR-211 was positively correlated with miR-202(r=0.651,P<0.05).According to Spearman correlation analysis,miR-211 and miR-202 were positively correlated with MMSE and MoCA(r=0.539～0.585,all P<0.05)and negatively correlated with HAMA and HAMD(r=-0.651～-0.539,all P<0.05).Logistic regression analysis showed that the low expression of miR-211[OR(95%CI):5.321(1.648～17.180)]and miR-202[OR(95%CI):3.158(1.989～5.012)]were risk factors for Alzheimer's disease(P<0.05).Conclusion The serum expression levels of miR-211 and miR-202 in patients with Alzheimer's disease were reduced,indicating miR-211 and miR-202 were closely related to cognitive function,anxiety and depression.

Objective:To evaluate the efficacy and safety of antaitasvir phosphate 100 mg or 200 mg combined with yiqibuvir for 12 weeks in patients with various genotypes of chronic hepatitis C, without cirrhosis or compensated stage cirrhosis.Methods:Patients with chronic hepatitis C (without cirrhosis or compensated stage cirrhosis) were randomly assigned to the antaitasvir phosphate 100 mg+yiqibuvir 600 mg group (100 mg group) or the antaitasvir phosphate 200 mg+yiqibuvir 600 mg group (200 mg group) in a 1∶1 ratio. The drugs were continuously administered once a day for 12 weeks and observed for 24 weeks after drug withdrawal. The drug safety profile was assessed concurrently with the observation of the sustained virological response (SVR12) in the two patient groups 12 weeks following the drug cessation. The intention-to-treat concept was used to define as closely as possible a full analysis set, including all randomized cases who received the experimental drug at least once. The safety set was collected from all subjects who received the experimental drug at least once (regardless of whether they participated in the randomization group) in this study. All efficacy endpoints and safety profile data were summarized using descriptive statistics. The primary efficacy endpoint was SVR12. The primary analysis was performed on a full analysis set. The frequency and proportion of cases were calculated in the experimental drug group (antaitasvir phosphate capsules combined with yiqibuvir tablets) that achieved "HCV RNA

Objective To investigate the correlation between serum Angiopoietin-like protein 4(ANGPTL4)and soluble bone marrow cell-like transcription factor-1(sTLT-1)levels and cognitive function and prognosis in elderly patients with vascular dementia(VD).Methods A total of 92 elderly patients with vascular dementia admitted to Hebei Yanda Hospital from June 2020 to June 2022 were selected as the study group.They were divided into mild(n=30),moderate(n=36)and severe groups(n=26)according to the minimum mental state examination(MMSE)score,and 92 patients with healthy physical examination during the same period were selected as the control group.According to the prognosis of patients,the cognitive dysfunction was divided into grade Ⅰ～Ⅱ and grade Ⅲ.ELISA method was used to detect the levels of serum ANGPTL4 and sTLT-1.Pearson and Spearman methods were used to analyze the correlation between serum ANGPTL4 and sTLT-1 and MOCA scores.Logistic regression analysis was used to analyze the influencing factors of the prognostic grade Ⅲ of elderly patients with vascular dementia.The receiver operating characteristic(ROC)curve was plotted to analyze the predictive value of serum ANGPTL4 and sTLT-1 levels in the prognostic grade Ⅲ in elderly patients with vascular dementia.Results The serum ANGPTL4 level(987.57±53.25 pg/ml)in the study group was lower than that in the control group(1 108.35±62.13 pg/ml),and the serum sTLT-1(68.01±5.15 pg/ml)level was higher than that in the control group(50.12±4.57 pg/ml),with significant differences(t=14.158,24.922,all P＜0.05).Serum ANGPTL4 levels and MOCA scores were decreased sequentially in the mild,moderate and severe groups(F=33.495,66.617),while serum sTLT-1 level was increased sequentially(F=66.718),and the differences were statistically significant(all P＜0.05),respective.Pearson analysis showed that serum ANGPTL4 was negatively correlated with sTLT-1(r=-0.621,P＜0.05).Spearman analysis showed serum levels of ANGPTL4 and sTLT-1 were positively and negatively correlated with MoCA scores,respectively(r=0.545,-0.557,all P＜0.05).The serum ANGPTL4 level(953.45±51.16 pg/ml)in the prognostic patients of grade Ⅲ was lower than that of grade Ⅰ～Ⅱ(1 005.76±54.27 pg/ml),and the serum level of sTLT-1(73.14±5.40 pg/ml)was higher than that of grade Ⅰ～Ⅱ(65.28±5.02 pg/ml),with significant differences(t=4.490,6.967,all P＜0.05).Logistic regression analysis showed low expression of ANGPTL4[OR(95％CI):5.089(1.833～14.129)]and high expression of sTLT-1[OR(95％CI):4.258(1.739～10.428)]were risk factors affecting the prognosis of elderly patients with vascular dementia(P＜0.05).According to the ROC curve,the combined prediction of two for the grade Ⅲ prognosis of elderly vascular dementia patients was better than the individual prediction of ANGPTL4 and sTLT-1(Z=2.135,3.268,all P＜0.05).Conclusion Serum ANGPTL4 level was decreased but sTLT-1 level was increased in elderly patients with vascular dementia,and ANGPTL4 and sTLT-1 were closely related to cognitive function and prognosis.

Objective:To investigate the expressions of tissue inhibitor of matrix metalloproteinase-1 (TIMP1) and fibronectin 1 (FN1) in pregnancy associated breast cancer (PABC) and their correlations with expression of E-cadherin (E-cad).Methods:The clinicopathological data of 55 PABC patients in Binzhou People's Hospital Affiliated to Shandong First Medical University from January 2011 to December 2020 were retrospectively analyzed. Immunohistochemistry was used to detect expressions of TIMP1, FN1 and E-cad in cancer tissues and corresponding paracancerous tissues (>3 cm from the edge of the tumor foci). The expressions of TIMP1 and FN1 proteins in fresh intraoperative frozen cancer tissues and paracancerous tissues of 10 PABC patients were detected by Western blotting. The correlations of TIMP1 and FN1 expressions with clinicopathological characteristics of patients were analyzed by χ2 test, the correlation of TIMP1 and FN1 expressions with E-cad expression was analyzed by Spearman method, and the correlation of TIMP1 and FN1 expressions with survival was analyzed by Kaplan-Meier method. Results:The positive rates of TIMP1 and FN1 in PABC tissues were 72.7% (40/55) and 58.2% (32/55), and 25.5% (14/55) and 18.2% (10/55) in paracancerous tissues, and the differences were statistically significant ( χ2 values were 24.59 and 18.64, both P < 0.001). The results of Western blotting showed that the relative expressions of TIMP1 and FN1 proteins in the fresh cancer tissues of 10 PABC patients was higher than those in the corresponding paracancerous tissues (1.60±0.76 vs. 0.62±0.29, 1.31±0.62 vs. 0.44±0.15), and the differences were statistically significant ( t values were 5.92 and 4.86, both P < 0.001). The expressions of TIMP1 and FN1 in PABC tissues were correlated with estrogen receptor expression, Ki-67 positivity index, TNM stage and lymph node metastasis (all P < 0.05). The expressions of TIMP1 and FN1 were negatively correlated with expression of E-cad in PABC ( r values were -0.471 and -0.432, both P < 0.001). Five cases were lost to follow-up, and the remaining 50 cases had a median follow-up time of 43 months (12-90 months). Among the 50 cases, 36 cases were TMP1-positive and 29 cases were FN1-positive. The overall survival of TIMP1-negative group and FN1-negative group were better than those of the corresponding positive group ( χ2 values were 4.49 and 6.06, both P < 0.05); the median overall survival time of TIMP1-positive group and FN1-positive group were 51 months (95% CI 37-65 months) and 43 months (95% CI 32-53 months), while that of TIMP1-negative group and FN1-negative group were 89 months (95% CI 84-93 months) and 87 months (95% CI 85-92 months). Conclusions:TIMP1 and FN1 expressions are elevated in PABC tissues and negatively correlated with E-cad expression, TIMP1 and FN1 may be involved in PABC invasion through epithelial-mesenchymal transition and affect the prognosis of patients.

Objective:To investigate the effect of the artificial facial nerve on the restoration of orbicularis oculi muscle function for unilateral peripheral facial paralysis in rabbit.Methods:Artificial facial nerve was implanted into the rabbit with unilateral peripheral facial paralysis between January 2018 and May 2019. At different time points after operation, the affected orbicularis oculi muscles′ stimulation closure threshold and the synchronism about the motion of the two sides of orbicularis oculi muscles were monitored. T test was used with SPSS 13.0 software. Results:There was no significant difference in the closure threshold of the orbicularis oculi muscle on the 7th and 28th days after artificial facial nerve implantation ( P>0.05). The synchronism of the normal side eye closing movement triggered the affected orbicular oculi muscle movement with the artificial facial nerve was up 19/20 to 20/20 at different times. Conclusions:The artificial facial nerve system had the features of stable working condition and high synchronizing effect for stimulating movement. It could restore the closed eye function in animals with peripheral facial paralysis animals and had great clinical application prospects.

Objective: To evaluate the efficacy and safety of sofosbuvir (Nanjing Zhengda Tianqing Pharmaceutical Co., Ltd.) combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection. Methods: Treatment-naïve or treatment experienced genotype 2 chronic hepatitis C patients from sixteen research centers of China were screened. All subjects received once-daily dose of sofosbuvir (400 mg) combined with ribavirin (body weight < 75 kg, 1 000 mg/day, 400 mg in the morning and 600 mg in the evening; body weight > 75 kg, 1 200 mg/d, 600 mg in the morning and 600 mg in the evening) for 12 weeks. Patients were followed-up for a period of 12 weeks after discontinuation of treatment. Continuous variables were expressed as mean ± standard deviation. The proportion of subjects with virologic response at different follow-up time points and 95% confidence intervals were estimated by maximum likelihood ratio and Clopper-Pearson interval. Results: 132 cases with genotype 2 chronic hepatitis C virus infection from sixteen research centers of China were included, 12 cases of whom were associated with cirrhosis, and the remaining 120 cases were not associated with cirrhosis. One hundred and thirty-one cases completed the study, and one patient lost to follow-up at week 4 after the end of treatment. The sustained virological response rate was 96.2% (95% confidence interval: 92.37% - 99.16%) after 12 weeks of drug withdrawal. Virological relapse occurred in four cases. Of the 132 subjects enrolled in the study, 119 (90.2%) reported 617 adverse events during treatment, of which 359 (76.5%) were TEAE related to sofosbuvir and/or ribavirin. There were nine TEAEs of grade 3 and above, and six cases (4.5%) of them had six severe adverse events. Only one serious adverse event was associated with sofosbuvir and ribavirin (unstable angina pectoris). There were no adverse events leading to drug discontinuation or death. Conclusion Sofosbuvir combined with ribavirin has a high SVR rate in the treatment of genotype 2 chronic hepatitis C virus infection, and most of the adverse events occurred were mild with acceptable safety profile.

OBJECTIVE: To explore the clinical, electrophysiological and imaging features of a patient with Krabbe disease caused by GALC mutation. METHODS: A comprehensive analysis including clinical investigation and genetic testing was carried out. RESULTS: The patient presented with peripheral neuropathy with electrophysiological anomaly suggestive of asymmetric demyelinating neuropathy. Brain imaging revealed leukoencephalopathy. Genetic analysis has identified compound heterozygous mutations in exons 5 and 11 of the GALC gene, namely c.461C>A and c.1244G>A. CONCLUSION Krabbe disease is a group of disorders featuring substantial phenotypic heterogeneity. Genetic and enzyme testing has become indispensable for accurate diagnosis for this disease.
DNA Mutational Analysis ; Galactosylceramidase ; genetics ; Genetic Testing ; Humans ; Leukodystrophy, Globoid Cell ; complications ; genetics ; Mutation ; Peripheral Nervous System Diseases ; etiology

Objective To analyze clinicopathological characteristics and the potential risk-related factors of female breast hyperplasia in different age groups.Method From Jan 2015 to Dec 2016,patients diagnosed with breast hyperplasia in 12 hospitals were evaluated.All patients completed the self-designed questionnaires on women'health,including basic demographic information,clinic examination information,radiologic information and pathologic results.The patients were divided into a young group (＜ 45 years old) and an elderly group (from 45 to 75 years old).Results There were 3 684 cases of breast hyperplasia,including 2 291 cases in young group and 1 393 cases in elder group,respectively Clinically breast pain type were most commonly observed in both young and older groups (50.3％ vs.42.7％,P ＜ 0.001).While pathological research based on biopsy showed that breast adenopathy were the most common changes in both groups (67.9％ vs.61.7％,P ＜0.001).More breast cancer cases were identified in elder group than that in young group,especially in clinically lump type patients (9.4％ vs.4.2％,P ＜ 0.001).Compared with elder group,patients in young group have different distribution characteristics regarding to fertility factors,lifestyle factors and psychology scale including anxiety and depression.Conclusion Distributions of clinicopathological characteristics and risk factors of female breast hyperplasia differ across different age groups.

OBJECTIVE： To establish a determination method for the concentration of cajanonic acid A （CAA） in liver microsome incubation system， and to compare the metabolism characteristics of it in different species of liver microsomes. METHODS： CAA was dissolved in liver microsome incubation system of rat， Beagle dog and human initiated by reduced nicotinamide adenine dinucleotide phosphate （NADPH）， and was incubated in water at 37 ℃. The reaction was terminated with acetonitrile at 0， 5， 10， 15， 30， 45 and 60 min， respectively. Using genistein as internal standard， the concentration of CAA in  different incubation systems was determined by UPLC-MS/MS. The determination was performed on Waters BEH C18 column with mobile phase consisted of water （containing 0.1％ formic acid）-acetonitrile （containing 0.1％ formic acid） （45 ∶ 55， V/V） at the flow rate of 0.25 mL/min. The column temperature was 30 ℃， and the sample size was 2 μL. The electrospray ionization source was used to the select reaction monitoring mode for negative ion scanning. The ion pairs for quantitative analysis were m/z 353.14→309.11 （CAA）， m/z 269.86→224.11 （internal standard） respectively. The residual percentage and enzymatic kinetic parameters of CAA in different incubation systems were calculated according to the mass concentration of CAA at 0 min. RESULTS： The linear range of CAA was 0.05-20 μg/mL； the limit of quanti- tation was 0.05 μg/mL， and the lowest detection limit was 0.01 μg/mL. RSDs of intra-day and inter-day were lower than 10％； relative errors ranged －4.83％-8.94％； extraction method and matrix effect did not affect the determination of the substance to be measured. At 60 min of incubation， residual percentages of CAA in rat， Beagle dog and human liver microsomes were（62.79±9.99）％，（64.07±11.59）％，（96.66±5.71）％， respectively. The half-life period （72.19， 68.61 min） of CAA in rat and Beagle dog liver microsomes were significantly shorter than human liver microsome （364.74 min）. The clearance rates [0.019 2， 0.020 2 mL/（min·mg）] were significantly higher than human liver microsome [0.003 8 mL/（min·mg）] （P＜0.05）. CONCLUSIONS： Established UPLC-MS/MS method is simple， rapid， specific and sensitive， and can be used for the determination of CAA concentration in liver microsome incubation system and the study of metabolism stability in vitro. The stability of CAA metabolism in rat and Beagle dog liver microsomes are poorer than human liver microsome.

A 28-year-old female patient presented with indurated erythema and nodules on the right lower limb for 2 years,with mild itching and pain.Skin examination showed a well-circumscribed irregular dark red patch measuring about 10 cm × 5 cm in size on the extensor aspect of the right thigh.On the patch,there were scattered or densely distributed mung bean-to soybean-sized quasi-circular violaceous nodules with a smooth surface,which were hard on palpation.Subcutaneous nodules with medium hardness were found on palpation,and hyperpigmentation was observed on the surface of some nodules.Local skin temperature was increased,with tenderness on palpation.Histopathologically,mononuclear cells showed nodular or sheet-like distribution in the middle and upper dermis,some of which had pale-staining cytoplasm.Moreover,plenty of plasma cells were observed.Immunohistochemistry revealed that histiocytes were stained strongly positive for S100.The number of IgG4-positive plasma cells increased obviously,and was more than 50 per high-power field (× 200).The proportion of IgG4-positive plasma cells in IgG-positive plasma cells was 45％.Finally,the patient was diagnosed with cutaneous Rosai-Dorfman disease with increased IgG4-positive plasma cells.