ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 （CCK-8） assay was employed to measure the viability of human ovarian granulosa cells （KGN cells） treated with Tusizi prescription extracts 1-11， and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP） and encyclopedia of traditional Chinese medicine （ETCM）. The common targets shared by Tusizi prescription and diminished ovarian reserve （DOR） were selected and imported into search tool for the retrieval of interacting genes/proteins （STRING） to construct a protein-protein interaction （PPI） network and into gene function annotation database （DAVID） for gene ontology （GO） analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay， 5-ethynyl-2'-deoxyuridine （EdU） staining， terminal-deoxynucleoitidyl transferase mediated nick-end labeling （TUNEL）， and enzyme-linked immunosorbent assay （ELISA） were employed to examine the proliferation， apoptosis， and estradiol （E₂） secretion of KGN cells treated with the water extract 11 of Tusizi prescription （Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1） and the optimal prescription screened by uniform design. On this basis， the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes， primarily involving cellular responses to exogenous compound stimuli， negative regulation of apoptotic process， and positive regulation of cell proliferation. It identified 84 cellular components， including cell membrane， mitochondria， and neuronal cell body， as well as 144 molecular functions such as enzyme binding， estrogen response element binding， and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen， Lycii Fructus， Dioscoreae Rhizoma， Poria， and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14， the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group， the hyperoside group demonstrated increased viability of ovarian germline stem cells （P<0.01）. The CCK-8， EdU， and ELISA results showed that compared with the normal group， the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells （P<0.05， P<0.01）. ELISA results showed that compared with the normal group， the water extract 11 of Tusizi prescription promoted the E₂ secretion of KGN cells （P<0.05）， while the optimal prescription screened by uniform design had no significant effect on the E₂ secretion. ConclusionBoth the water extract 11 of Tusizi prescription （Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1） and the optimal prescription screened by uniform design （Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14） can improve the ovarian reserve function， and the former has better effect. Tusizi prescription can modulate biological processes （such as cell proliferation and apoptosis） and molecular functions （such as enzyme binding and estrogen response element binding） through active components like hyperoside to promote the proliferation and E₂ secretion and inhibit the apoptosis of KGN cells， thereby protecting the ovarian reserve function.

ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 （CCK-8） assay was employed to measure the viability of human ovarian granulosa cells （KGN cells） treated with Tusizi prescription extracts 1-11， and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP） and encyclopedia of traditional Chinese medicine （ETCM）. The common targets shared by Tusizi prescription and diminished ovarian reserve （DOR） were selected and imported into search tool for the retrieval of interacting genes/proteins （STRING） to construct a protein-protein interaction （PPI） network and into gene function annotation database （DAVID） for gene ontology （GO） analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay， 5-ethynyl-2'-deoxyuridine （EdU） staining， terminal-deoxynucleoitidyl transferase mediated nick-end labeling （TUNEL）， and enzyme-linked immunosorbent assay （ELISA） were employed to examine the proliferation， apoptosis， and estradiol （E₂） secretion of KGN cells treated with the water extract 11 of Tusizi prescription （Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1） and the optimal prescription screened by uniform design. On this basis， the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes， primarily involving cellular responses to exogenous compound stimuli， negative regulation of apoptotic process， and positive regulation of cell proliferation. It identified 84 cellular components， including cell membrane， mitochondria， and neuronal cell body， as well as 144 molecular functions such as enzyme binding， estrogen response element binding， and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen， Lycii Fructus， Dioscoreae Rhizoma， Poria， and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14， the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group， the hyperoside group demonstrated increased viability of ovarian germline stem cells （P<0.01）. The CCK-8， EdU， and ELISA results showed that compared with the normal group， the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells （P<0.05， P<0.01）. ELISA results showed that compared with the normal group， the water extract 11 of Tusizi prescription promoted the E₂ secretion of KGN cells （P<0.05）， while the optimal prescription screened by uniform design had no significant effect on the E₂ secretion. ConclusionBoth the water extract 11 of Tusizi prescription （Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1） and the optimal prescription screened by uniform design （Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14） can improve the ovarian reserve function， and the former has better effect. Tusizi prescription can modulate biological processes （such as cell proliferation and apoptosis） and molecular functions （such as enzyme binding and estrogen response element binding） through active components like hyperoside to promote the proliferation and E₂ secretion and inhibit the apoptosis of KGN cells， thereby protecting the ovarian reserve function.

Humans ; Arthroplasty, Replacement, Knee/adverse effects* ; Surgical Wound Infection/etiology* ; Retrospective Studies ; Risk Factors ; Arthroplasty, Replacement, Hip/adverse effects* ; Prosthesis-Related Infections/etiology*

OBJECTIVE: To determine the composition and distribution of beta-thalassemia-associated genotypes in Liuzhou area of Guangxi, China. METHODS: From January to December 2017, 13 847 individuals who came for premarital examination, maternity examination or health check were recruited with informed consent. The subjects were analyzed by reverse dot blotting (RDB) for 17 common beta-thalassemia-associated variants among the Chinese population. Individuals with inconsistent results by blood test, electrophoresis, and RDB were subjected to Sanger sequencing to detect rare variants of the beta globin gene. RESULTS: In total 2098 individuals were found to harbor beta-thalassemia-associated variants, which included 2075 heterozygotes (98.90%), 12 compound heterozygotes (0.57%) and 11 homozygotes (0.52%). CD41-42 (48.43%) and CD17 (31.45%) were the most common variants. Three hundred and thirty eight-individuals were found to also carry heterozygous variants of the alpha globin gene, with the most common types being --SEA/aa, -a3.7/aa, aCSa/aa, -a4.2/aa. Through Sanger sequencing, rare genotypes such as beta-32/betaN, betaCD41-42/betaIVS-II-5 and betaCD30/betaN were detected. CONCLUSION Liuzhou area has a high incidence of beta-thalassemia, but with a complex variant spectrum and clinical phenotypes different from other regions. Genetic counseling and prenatal diagnosis for the carrier population is crucial for the reduction of the related birth defects. Our result may provide valuable information for the prevention and control of beta-thalassemia in this area.
China ; Female ; Genetic Counseling ; Genetic Variation ; Genotype ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; alpha-Globins ; genetics ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; genetics

Objective? To explore the correlation between preoperative cognitive function and anxiety and depression in patients with laryngocarcinoma. Methods? Totally 42 patients with laryngocarcinoma who were hospitalized in the Department of Otorhinolaryngology Head and Neck Surgery of 2 ClassⅢ Grade A Hospitals in Shanxi Province from September 2017 to September 2018 were selected into the observation group by convenient sampling, while 40 healthy volunteers were included in the control group. They were investigated with Self-Rating Anxiety Scale (SAS),Self-Rating Depression Scale (SDS) and Montreal Cognitive Assessment (MOCA). Results? The preoperative SAS and SDS scores of the observation group were (41.48±5.46) and (43.69±6.16) respectively, both higher than those of the control group (t=4.189, 6.234; P＜ 0.01); the preoperative MOCA score of the observation group was (22.90±4.13), lower than that of the control group (t=2.646, P＜ 0.01). Correlation analysis showed that the laryngocarcinoma patients＇ anxiety and depression were negatively correlated with their cognitive function (r=-0.750, -0.660; P＜0.01). Conclusions? Compared with the healthy volunteers, the laryngocarcinoma patients are more susceptible to anxiety, depression and cognitive disorder. The severer their anxiety and depression, the poorer their overall cognitive function was.

BACKGROUND: A large number of cytokines produce in the periodontium on the tension side during orthodontic tooth movement, participate in periodontal remodeling, and make the tooth move to its normal position, thus achieving alveolar remodeling. Heat shock protein 27 (HSP27) is reported to be involved in the periodontal remodeling. OBJECTIVE: To investigate the expression of HSP27 in the rat periodontium on the tension side during the orthodontic tooth movement.METHODS: The rat models of orthodontic tooth movement were constructed using NiTi coil spring with the force value of 50 g. The periodontal samples were collected at 1, 3, 5, 7, 10 and 14 days after exerting force to detect the expression level of HSP27 in the periodontium on the tension side.RESULTS AND CONCLUSION: Immunofluorescence staining results revealed that the expression level of HSP27 in the periodontal tissues began to increase at the 1st day after exerting force, reached the highest level at the 3rd day, and then fell to the initial level at the 5th day. To conclude, HSP27 is evoked by orthodontic force early, and it may play a role in periodontal remodeling.

BACKGROUND:Extensive research has been concentrated on the reconstruction of periodontium under orthodontic force. However, there are no reports regarding the expression of heat shock protein 70 (HSP70) in rat periodontium during orthodontic tooth movement. OBJECTIVE:To investigate the expression of HSP70 in rat periodontium during orthodontic tooth movement. METHODS:Rat orthodontic tooth movement models were established using NiTi extension spring with the force of 50 g. The orthodontic periodontium was col ected at 1, 3, 5, 7, 10 and 14 days after exerting force to detect the expression of HSP70. RESULTS AND CONCLUSION:According to immunofluorescence staining, the expression of HSP70 in the periodontal tissues was increased on day 1 after exerting force, and reached the highest level on day 5, then decreased to the initial level on day 14. These results suggest that the expression of HSP70 is evoked by orthodontic force, and it may play a role in periodontal remodeling.

Objective To systematically evaluate the association between lactotransferrin(LTF) gene rs1126478 polymor‐phism and caries susceptibility .Methods The published literatures on the association between LTF gene rs1126478 polymorphism and caries susceptibility were retrived from the electronic databases of Pubmed ,Web of science ,Embase ,EBSCO ,Springerlink , CNKI ,VIP ,Wanfang and CBM by computer .The pooled odds ratios (ORs) and its 95% confidence intervals (95% CIs) were used as the effect indicators .The statistical analysis was performed by using the RevMan 5 .2 and Stata 12 .0 softwares .The bias evalua‐tion and sensitivity analysis were also performed .Results A total of 5 case‐control studies involving 720 cases and 412 controls were included .The meta analysis results showed that no statistical significance in the association between LTF gene rs1126478 pol‐ymorphism and caries susceptibility was revealed in all four genetic models (GG+ AG vs .AA :P= 0 .75 ,OR= 0 .93 ,95% CI=0 .59-1 .46 ;G vs .A :P=0 .88 ,OR=0 .97 ,95% CI=0 .68 -1 .38 ;GG vs .AA :P=0 .84 ,OR=1 .07 ,95% CI =0 .57 -1 .99 ;GG vs .AG+AA :P=0 .52 ,OR=1 .12 ,95% CI = 0 .79-1 .58) .Similarly ,the subgroup analysis by ethnicity showed no statistical sig‐nificance in the onset risk between Caucasian and Asian populations as well (both P>0 .05) .Conclusion The LTF gene rs1126478 polymorphism may have no relation with the susceptibility to caries .

Objective To establish the quality standard of Radix Toddaliae Asiaticae. Methods Thin layer chromatography ( TLC) and high performance liquid chromatography ( HPLC) were used to identify and determine chloride nitidine and toddalolactone in Radix Toddaliae Asiaticae. The moisture and total ash contents were detected according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition) . Results Toddalolactone and chloride nitidine were detectable by TLC, the spots were clear and the dissociation was good. The established HPLC method was simple and accurate. The linear ranges of toddalolactone and chloride nitidine in Radix Toddaliae Asiaticae were 2.84~42.6 μg/mL and 25.6~385 μg/mL, and their recovery rates were 99.2 % ( RSD=1.12%) and 100 % ( RSD=0.71%) , respectively. The content of moisture was in the range of 75.8~98.9 mg/g and that of total ash was in the range of 12.4~33.6 mg/g. Conclusion The developed method is specific and accurate, and can provide useful reference for establishing quality standard of Radix Toddaliae Asiaticae.

Objective To observe the change of oxidalive stress indexes in the patients with diabetic perineuropathy (DPN), and investigate the protection role of glutathione (GSH) on neuroprotective effect. Methods The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) and sensory nerve conduction velocity (SCV) were detected in 54 cases with DPN (DPN group) and 30 cases of health examination (control group). Fifty-four cases with DPN were divided into GSH group (given basic treatment and GSH 1.8 g/d) and CON group(given basic treatment and B vitamin) with 27 cases each by random-digits table. After treatment of 2 weeks, the levels of SOD, MDA and SCV were compared. Results The levels of MDA in DPN group were significantly higher than those in control group, while the activity of SOD was significantly lower than that in control group [(7.23 ±2.31) μmol/L vs. (4.87 ± 1.17) μmol/L, (59.72 ± 13.58) kU/L vs. (76.19 ± 7.55 ) kU/L](P＜ 0.01). After treatment, the level of MDA was decreased and the activity of SOD was increased in GSH group and CON group (P ＜0.05),but there was no significant difference between the two groups(P＞ 0.05). After treatment, the level of SCV was significantly increased in GSH group,while there was no significant difference in CON group (P ＞0.05). Conclusions Oxidative stress exists in DPN patients. The antioxidant treatment with GSH can improve impaired nerve and has a better effect of nerve protection than classical nerve nutrition therapy.