Objective To study the effect of Liuwei Dihuangwan on autophagy level and its mechanism in SAMP8 mice and Aβ-stimulated BV2 cell model,and to explore the molecular mechanism of tonifying the kidney and filling up the essence to prevent and control Alzheimer's disease(AD)through interfering with autophagy.Methods Ten 7-month-old male anti-aging mice(SAMR1)were taken as the normal group,and 40 7-month-old male rapid aging mice(SAMP8)were randomly control and model groups,equal volumes of saline were administered by gavage twice a day for 4 weeks,and the levels of Aβ expression in the hippocampus of the mice in each group were detected by immunofluorescence;The expression levels of FcγRⅡB,c-Src and SHP-1 in the hippocampus of each group were detected by Western blot;BV2 cells were cultured and Fcγ receptor Ⅱ-b(FcγRⅡB)overexpression vectors were constructed;the AD state cell model was established by treating the BV2 cells with 5 μmol·L-1 Aβ1-42,and the Liuwei Dihuangwan drug-containing serum was prepared.The cells were divided into NC group,Aβ1-42 group,blank serum group,drug-containing serum group,Vector group,FcγRⅡB OE group,and drug-containing serum+FcγRⅡB OE group;immunofluorescence was used to detect the expression level of Aβ protein in the cells of each group;Western blot was used to detect the expression level of p62,LC3 Ⅱ/Ⅰ,FcγRⅡB,SHP-1,and c-Src in cells of each group.Results Compared with the normal group,the hippocampal Aβ,FcγRⅡB,SHP-1,and c-Src expression levels in the model group of mice were significantly higher(P<0.01),and compared with the model group,the expression levels of Aβ,FcγRⅡB,SHP-1,and c-Src in the low-,medium-,and high-dose groups of Liuwei Dihuangwan were significantly lower(P<0.01),it also showed a significant dose dependent relationship.Compared with NC group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in Aβ1-42 group were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with Aβ1-42 group and blank serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in drug-containing serum group were significantly decreased(P<0.01),and LC3Ⅱ/Ⅰ was significantly increased(P<0.01);Compared with NC group and Vector group,the expression of Aβ in FcγRⅡB OE group was increased,the protein expressions of p62,FcγRⅡB,SHP-1 and c-Src were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with the drug-containing serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in the drug-containing serum+FcγRⅡB OE group were significantly increased(P<0.01),and the protein expression levels of LC3Ⅱ/Ⅰ were significantly decreased(P<0.01).Conclusion Liuwei Dihuangwan improved AD by inhibiting microglia FcγRⅡB/c-Src pathway and increasing autophagy level.

Objective To study the effect of Liuwei Dihuangwan on autophagy level and its mechanism in SAMP8 mice and Aβ-stimulated BV2 cell model,and to explore the molecular mechanism of tonifying the kidney and filling up the essence to prevent and control Alzheimer's disease(AD)through interfering with autophagy.Methods Ten 7-month-old male anti-aging mice(SAMR1)were taken as the normal group,and 40 7-month-old male rapid aging mice(SAMP8)were randomly control and model groups,equal volumes of saline were administered by gavage twice a day for 4 weeks,and the levels of Aβ expression in the hippocampus of the mice in each group were detected by immunofluorescence;The expression levels of FcγRⅡB,c-Src and SHP-1 in the hippocampus of each group were detected by Western blot;BV2 cells were cultured and Fcγ receptor Ⅱ-b(FcγRⅡB)overexpression vectors were constructed;the AD state cell model was established by treating the BV2 cells with 5 μmol·L-1 Aβ1-42,and the Liuwei Dihuangwan drug-containing serum was prepared.The cells were divided into NC group,Aβ1-42 group,blank serum group,drug-containing serum group,Vector group,FcγRⅡB OE group,and drug-containing serum+FcγRⅡB OE group;immunofluorescence was used to detect the expression level of Aβ protein in the cells of each group;Western blot was used to detect the expression level of p62,LC3 Ⅱ/Ⅰ,FcγRⅡB,SHP-1,and c-Src in cells of each group.Results Compared with the normal group,the hippocampal Aβ,FcγRⅡB,SHP-1,and c-Src expression levels in the model group of mice were significantly higher(P<0.01),and compared with the model group,the expression levels of Aβ,FcγRⅡB,SHP-1,and c-Src in the low-,medium-,and high-dose groups of Liuwei Dihuangwan were significantly lower(P<0.01),it also showed a significant dose dependent relationship.Compared with NC group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in Aβ1-42 group were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with Aβ1-42 group and blank serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in drug-containing serum group were significantly decreased(P<0.01),and LC3Ⅱ/Ⅰ was significantly increased(P<0.01);Compared with NC group and Vector group,the expression of Aβ in FcγRⅡB OE group was increased,the protein expressions of p62,FcγRⅡB,SHP-1 and c-Src were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with the drug-containing serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in the drug-containing serum+FcγRⅡB OE group were significantly increased(P<0.01),and the protein expression levels of LC3Ⅱ/Ⅰ were significantly decreased(P<0.01).Conclusion Liuwei Dihuangwan improved AD by inhibiting microglia FcγRⅡB/c-Src pathway and increasing autophagy level.

Objective:To explore the effects of Liuwei Dihuang Wan on the behaviors and Toll-like receptor 4/nuclear factor kappa-B(TLR4/NF-κB) signal transduction pathway of amyloid β-precursor protein/presenilin-1(APP/PS1) double transgenic mice.Methods:Forty 3-month-old female APP/PS1 mice were randomly divided into model group, low-dose group(0.59 g/kg), medium-dose group(1.18 g/kg), high-dose group(2.36 g/kg)of Liuwei Dihuang Wan(gavaged according to grouped doses), and ibuprofen group(0.04 g/kg, gavage) using a random number table method, with 8 mice in each group.Eight 3-month-old wild-type female C57BL/6 mice with matched body weight were used as the control group.The mice in control group and model group were given an equal volume of 0.9% sodium chloride solution by gavage.The gavage administration was twice a day for a continuous period of 3 months.Morris water maze test was used to detect the learning and memory abilities of mice. ELISA was used to detect the serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and immunohistochemistry was used to detect the levels of amyloid β-protein (Aβ), glial fibrillary acidic protein(GFAP) and NF-κB in hippocampal tissue.Western blot was used to detect the expression levels of TLR4, myeloid differentiation primary response gene 88(MyD88), and phosphorylated NF-κB(p-NF-κB) proteins in hippocampal tissue.The SPSS 20.0 software was used for data analysis. Multiple group comparisons were conducted by repeated measure ANOVA or one-way ANOVA.Results:The results of repeated measure ANOVA showed that as for the escape latency of the 6 groups of mice, the interaction effect between time and group was significant ( Finteraction=117.219, P<0.001). The escape latencies of mice in the 6 groups on the 5th day were all lower than those on the 1st day (all P<0.05). The escape latencies of mice in the ibuprofen group and the medium-dose and high-dose groups of Liuwei Dihuang Wan were lower than that in the model group from 1st day to 5th day(all P<0.05). On the 3rd to 5th day, the escape latencies of mice in the medium-dose and high-dose groups of Liuwei Dihuang Wan were lower than those in the low-dose group of Liuwei Dihuang Wan (all P<0.05). There were statistically significant differences in the percentage of residence time in the platform quadrant and the numbers of crossing platform among the 6 groups of mice ( F=5.451, 4.824, both P<0.05). The percentage of residence time in the platform quadrant (50.77±5.49)%, (54.39±5.71)%, (51.98±6.12)%), and the numbers of crossing platform((5.9±1.1) times, (6.0±1.3) times, (5.1±0.8) times) in the high-dose and medium-dose groups of Liuwei Dihuang Wan and the ibuprofen group were all higher than those in the model group ((27.32±3.22)%, (2.2±1.0) times )(all P<0.05). The immunohistochemical results showed that there were statistically significant differences in the integrated optical density values of Aβ, GFAP and NF-κB in the hippocampal tissues of 6 groups of mice ( F=57.52, 45.37, 79.10, all P<0.05). The integrated optical density values of Aβ, GFAP and NF-κB in the high-dose and medium-dose groups of Liuwei Dihuang Wan and the ibuprofen group were all lower than those in the model group (all P<0.05). And the integrated optical density values of Aβ, GFAP, and NF-κB in the high-dose and medium-dose groups of Liuwei Dihuang Wan were all lower than those in the low-dose group of Liuwei Dihuang Wan (all P<0.05). There were statistically significant differences in the levels of serum TNF-α and IL-1β detected by ELISA ( F=3.996, 6.395, both P<0.05) and the proteins levels of TLR4, MyD88, and p-NF-κB in hippocampal tissue detected by Western blot among the 6 groups( F=15.710, 3.522, 4.119, all P<0.05). The serum TNF-α and IL-1β levels in the high-dose and medium-dose groups of Liuwei Dihuang Wan and ibuprofen group were all lower than those in the model group (all P<0.05). The serum TNF-α ((18.90±2.33) ng/L, (21.56±2.49) ng/L) and IL-1β ((5.88±0.80) ng/L, (6.75±0.83) ng/L) levels in the high-dose and medium-dose groups of Liuwei Dihuang Wan were lower than those in the low-dose group ((30.77±2.89) ng/L, (9.11±1.27) ng/L) (all P<0.05). The protein expression levels of TLR4, MyD88, and p-NF-κB in the hippocampus of the high-dose and medium-dose groups of Liuwei Dihuang Wan and ibuprofen group were lower than those of the model group (all P<0.05). The protein expression levels of TLR4 ((0.254±0.091), (0.318±0.122)), MyD88 ((0.229±0.077), (0.386±0.119)), and p-NF-κB ((0.412±0.188), (0.358±0.119)) in the hippocampus of the high-dose and medium-dose groups of Liuwei Dihuang Wan were lower than those of the low-dose group ((0.617±0.172), (0.672±0.166), (0.799±0.227)) (all P<0.05). Conclusion:Liuwei Dihuang Wan can significantly alleviate learning and memory impairment in Alzheimer's disease model mice, possibly by inhibiting TLR4/NF-κB signal pathway, reducing TNF-α and IL-1β expression, thereby alleviate central immune inflammatory response and exert anti Alzheimer's disease effects.

Objective:To construct a targeted and accurate evaluation system for facial and cervical wounds and scars of burn patients.Methods:The method combining literature analysis and survey research was adopted, and the basic principles of item system construction were followed. From June to August 2020, based on the aesthetic standards of facial and cervical plastic surgery, the topographic map assessment system for facial and cervical wounds and scars of burn patients was preliminarily formed, focusing on the assessment of wounds and scars in the necks and faces of patients after burns. In September 2020, 38 experts in the relevant fields were consulted in advance and the questionnaire was revised according to the experts' opinions. From December 2020 to March 2021, the Delphi method was applied to conduct inquiry by correspondence with 35 experts in relevant fields from Guangzhou, Shenzhen, Shanghai, Beijing, and other cities, who met the inclusion criteria, and the items were screened and established. The effective recovery rate of inquiry questionnaire was calculated to determine the level of enthusiasm of experts, the average authority coefficient of all items was calculated to determine the level of expert authority, the average importance expert score, the average coefficient of variation, and the average full score rate of all the third-level items were calculated to determine the concentration of expert opinions, the average coefficients of variation and Kendall's harmony coefficients of the importance, sensitivity, and operability expert scores of all the third-level items were calculated to determine the degree of coordination of expert opinions. The Kendall's harmony coefficients for the importance, sensitivity, and operability expert scores of all the third-level items were statistically analyzed with chi-square test.Results:Among the 35 experts consulted by Delphi method, mainly were male, aged (48±10) years, with 8-38 years of working experience, mainly with associate senior titles and above, all with a bachelor's degree or above education background, and of whom 11 were burn experts, 7 were wound repair experts, 4 were plastic surgery experts, and 13 were rehabilitation medicine experts. Finally, a topographic map assessment system for facial and cervical wounds and scars of burn patients was formed, including 4 first-level items, 21 second-level items, 40 third-level items, and 1 mask. The effective recovery rate of inquiry questionnaire was 100% (35/35). The average authority coefficient of all items was 0.89. The average importance expert score was 4.67, the average coefficient of variation of importance expert score was 0.01, and the average full score rate of all the third-level items was 86.3%. The average coefficients of variation of the importance, sensitivity, and operability expert scores of all the third-level items were 0.01, 0.01, and 0.02, respectively. The Kendall's harmony coefficients for the importance, sensitivity, and operability expert scores of all the third-level items were statistically significant (with χ2 values of 1 201.53, 745.67, and 707.07, respectively , P<0.05). Conclusions:The established topographic map assessment system for facial and cervical wounds and scars of burn patients has high scientificity and reliability, which can be used for the evaluation of facial and neck wounds or scars in burn patients.

Objective:To study the trend of menarche age in Han and Mongolian women born from 1951 to 2005 in Mongolian region.Methods:A cross-sectional cluster sampling survey method was adopted, From 2003 to 2019, a retrospective survey was carried out in three banners/counties in Tongliao region on the female population of Han and Mongols nationalities aged 16 to 46 and conducted under standardized survey procedures and quality control standards. The basic data of menarche age of women born between 1951 and 2005 were obtained. The changes and rules were analyzed by taking 1 year, 5 years and 10 years as nodes.Results:Totally 46 and conducted under standardized survey procedures and quality control standards 928 pepole (24 450 Han and 22 478 Mongolian) were recruited, the survey response rate was 96.09% (46 928/48 836). In one-year-period analysis, the menarche age gradually decreased from 1951 to 2005. The mean menarche age of Han and Mongolian women changed from (16.22±0.52) years and (15.86±1.24) years in 1951 to (12.37±1.15) years and (12.33±0.98) years in 2005, respectively. The mean menarche age of Han and Mongolian women decreased 3.85 years and 3.54 years. The trend of the mean menarche age's change showed a significant negative correlation with the years (all P<0.000 1). In five-year-period analysis, the mean menarche age of Han and Mongolian women changed from (15.54±1.45) years and (15.53±1.48) years from 1951 to 1955 to (12.41±0.97) years and (12.47±0.96) years from 2001 to 2005, the mean menarche age decreased 3.13 years (3.41 months ahead of schedule every 5 years on average) and 3.06 years (3.34 months ahead of schedule every 5 years on average) in Han and Mongolian women respectively. In ten-year-period analysis, the mean menarche age of Han and Mongolian women changed from (15.79±0.95) years and (15.53±1.33) years from 1951 to 1960 to (12.41±0.97) years and (12.47±0.96) years from 2001 to 2005, the mean menarche age decreased 3.38 years (6.76 months ahead of schedule every 10 years on average) and 3.06 years (6.12 months ahead of schedule every 10 years on average) in Han and Mongolian women respectively. During the 15 years from 1951 to 1965, 1966 to 1970, 1971 to 1990, and 1991 to 2000, they were concentrated at the ages of 15-16, 14-15, 13-14, and 12-13, respectively. The proportion of women at 11 years, 12 years and 13 years menarche age were 26.79% (457/1 706), 73.27% (1 250/1 706), and 92.85% (1 584/1 706) during 2001—2005 in Han women, while the proportion were 23.25% (653/2 809), 62.01% (1 742/2 809), and 90.14% (2 532/2 809) in Mongolian women. Conclusion:The menarche age decreased in Han and Mongolian women from 1951 to 2005, and the ethnic groups tended to be the same. It is recommended to start adolescent education at the age of 8-9 years and pay attention to the changing pattern of early onset of menarche.

Objective:To study the trend of menarche age in Han and Mongolian women born from 1951 to 2005 in Mongolian region.Methods:A cross-sectional cluster sampling survey method was adopted, From 2003 to 2019, a retrospective survey was carried out in three banners/counties in Tongliao region on the female population of Han and Mongols nationalities aged 16 to 46 and conducted under standardized survey procedures and quality control standards. The basic data of menarche age of women born between 1951 and 2005 were obtained. The changes and rules were analyzed by taking 1 year, 5 years and 10 years as nodes.Results:Totally 46 and conducted under standardized survey procedures and quality control standards 928 pepole (24 450 Han and 22 478 Mongolian) were recruited, the survey response rate was 96.09% (46 928/48 836). In one-year-period analysis, the menarche age gradually decreased from 1951 to 2005. The mean menarche age of Han and Mongolian women changed from (16.22±0.52) years and (15.86±1.24) years in 1951 to (12.37±1.15) years and (12.33±0.98) years in 2005, respectively. The mean menarche age of Han and Mongolian women decreased 3.85 years and 3.54 years. The trend of the mean menarche age's change showed a significant negative correlation with the years (all P<0.000 1). In five-year-period analysis, the mean menarche age of Han and Mongolian women changed from (15.54±1.45) years and (15.53±1.48) years from 1951 to 1955 to (12.41±0.97) years and (12.47±0.96) years from 2001 to 2005, the mean menarche age decreased 3.13 years (3.41 months ahead of schedule every 5 years on average) and 3.06 years (3.34 months ahead of schedule every 5 years on average) in Han and Mongolian women respectively. In ten-year-period analysis, the mean menarche age of Han and Mongolian women changed from (15.79±0.95) years and (15.53±1.33) years from 1951 to 1960 to (12.41±0.97) years and (12.47±0.96) years from 2001 to 2005, the mean menarche age decreased 3.38 years (6.76 months ahead of schedule every 10 years on average) and 3.06 years (6.12 months ahead of schedule every 10 years on average) in Han and Mongolian women respectively. During the 15 years from 1951 to 1965, 1966 to 1970, 1971 to 1990, and 1991 to 2000, they were concentrated at the ages of 15-16, 14-15, 13-14, and 12-13, respectively. The proportion of women at 11 years, 12 years and 13 years menarche age were 26.79% (457/1 706), 73.27% (1 250/1 706), and 92.85% (1 584/1 706) during 2001—2005 in Han women, while the proportion were 23.25% (653/2 809), 62.01% (1 742/2 809), and 90.14% (2 532/2 809) in Mongolian women. Conclusion:The menarche age decreased in Han and Mongolian women from 1951 to 2005, and the ethnic groups tended to be the same. It is recommended to start adolescent education at the age of 8-9 years and pay attention to the changing pattern of early onset of menarche.

Objective:To investigate the effect of Lee Silverman voice therapy (LSVT) on the speech and life quality of Chinese persons with Parkinson′s disease (PD).Methods:A total of 16 Chinese PD patients were enrolled and given standard LSVT for 4 weeks. Their acoustic data were analyzed using PRAAT software before and after the treatment. Their voice quality was evaluated using the Japanese GRBAS voice scale and their quality of life was quantified using the Voice Handicap Index (VHI) scale.Results:After 4 weeks of LSVT the patients′ average voice and life quality had improved significantly. The average maximum duration of sustained vowel phonation had increased significantly, as had the vowels′ mean loudness, reading and monologue delivery. The GRBAS grading indicated that hoarseness had decreased significantly. The average total voice handicap index had also decreased significantly.Conclusion:Lee Silverman therapy can significantly improve maximum phonation time and loudness of Chinese-speakers, which enhances sensory feedback. The quality of their speech and of their life also improve significantly. This technique is recommended for clinical application.

ObjectiveTo evaluate the effects of dexmedetomidine on the quality of emergence from general anesthesia in the elderly patients undergoing orthopedic operation.MethodsSixty ASA Ⅰ -Ⅲ patients,aged ≥65 yr,undergoing elective orthopedic operation,were randomly assigned to one of 3 groups ( n =20 each):control group (group C) ; dexmedetomidine 0.25 μg/kg group (group D1 ) and dexmnedetomidine 0.50 μg/kg group (group D2).In groups D1 and D2,15 min befone anesthesia induction,dexmedetomidine 0.25 μg/kg and 0.50 μg/kg were infused over 15 min respectively,while the equal volume of normal saline 15 ml was given instead of dexmedetomidine in group C.After anesthesia induction,tracheal intubation was performed and the patients were mechanically ventilated.General anesthesia was used in the three groups.Flurbiprofen 1 mg/kg was injected intravenously immediately before skin incision.The time for recovery of spontaneous breathing,emergence time,extubation time,and adverse reactions were recorded.Pain was assessed with verbal rating scale (VRS) at 5 min after emergence from anesthesia.ResultsCompared with group C,the incidence of agitation,bucking,and adverse cardiovascular events and VRS score were significantly decreased,and the rate of effective analgesia was significantly increased in groups D1 and D2,and the emergence time and extubation time were significantly prolonged in group D2(P ＜0.05).Compared with group D1,the emergence time and extubation time were significantly prolonged in group D2(P＜0.05).There was no significant difference in VRS score,the rate of effective analgesia and the incidence of adverse renctions between groups D1 and D2(P＞0.05).ConclusionⅣ infusion of dexmedetomidine 0.25 μg/kg before induction of general anesthesia can effectively improve the quality of emergence from general anesthesia in the elderly patients undergoing orthopedic operation.