Objective: To investigate the effects of clinical therapeutic and pharmacological actions “Shenkang Oral Liquid” on sexual function failing, senile nocturia. Results: Index of phagocyte, weight of seminal vesicle gland and content of testosterone in experimental rats with yang deficiency were observed by experiment.Results: “Shenkang Oral Liquid” could significantly raise index of phagocyte (k=0.0258?0.012), weight of seminal vesicle gland (24.07?1.99mg/10g) and content of testosterone (363?50.56pmol/mL) in rats with yang deficiency. Conclusion: “Shenkang Oral Liquid” shows enhancement effects on immune and sexual functions.

Objective To investigate the air pollution level and the distribution regularity of air pollutants in Jilin City. Methods Based on the data from air quality automatic monitoring system in environmental protection station in Jilin City from 2001 to 2005, the concentration change of the inhalable particulate matter (PM10), SO2, NO2 and TSP in the air were analyzed with hour, season and year in different functional areas. Results The concentration of SO2, NO2 in atmosphere in winter and summer showed a daily changing with a bimodal curve, which appeared a high value section at 19:00—21:00 and 7:00—8:00. The highest concentration of pollutants appeared in winter, the lowest one in summer. The daily average concentration range of SO2, NO2 and TSP was 0.021-0.045, 0.027-0.046 and 0.300-0.470 mg/m3 respectively from 2001 to 2005 with a descending trend by years. As for the space distribution, the pollution in the industry area was the heaviest. Conclusion The air pollution type in Jilin City is still a coal-burning pollution, the concentration of air pollutants distributes differently with time and space.

Objective The study aimed to analyze the clinical data of patients with pulmonary embolism and summarize the clinical characteristics of pulmonary embolism,in order to improve the correct diagnostic rate and the prognosis. Methods Data of 178 patients who were cured in our hospital from January 2011 to December 2014 were analyzed retrospectively,including relative risk factors,clinical behaviors,examination data,misdiagnosis and prognosis. Results Patients with pulmonary embolism had related basic diseases and risk factors. Their clinical symptoms,signs and ordinary detection methods indicated pneumatorexis and cardiovascular injury,but lacked of specificity. The misdiagnosis rate of the first diagnosis was 48.88%,including 22 cases of the misdiagnosis to COPD, 18 cases to pneumonia and 16 cases to coronary heart disease. The misdiagnosis rate of elderly patients was 56.36%, while that of the young and middle-aged patients was 36.80%,the difference was statistically significant,P < 0.05. Computed tomography pulmonary angiography(CTPA)could accurately observe the pathological changes of pulmonary embolism,and all the patients were diagnosed by CTPA in the research. 11 cases(10%)of elderly patients died,while 2 cases(2.9%)happened in young and middle-aged patients. There was no statistical significant difference(P > 0.05). Conclusion Although the related risk factors of pulmonary embolism in patients existed,the misdiagnosed rate of the disease was high and prognosis was poor due to many confusing basic diseases. And the symptoms,signs and ordinary inspection of the patients lacked specificity. CTPA,being utilized to diagnose pulmonary embolism reliably,should be promoted.

Objective To determine the relationship between CCDC8 gene and breast cancer.Methods 40 cancerous breast tissue and 22 benign breast tissue were included.qRT-PCR was performed to investigate the expression level of CCDC8 in breast tissue.The correlation between CCDC8 level and the age of patients,tumor size,clinical staging,and the expression levels of estrogen and progesterone receptors,CerbB2,Ki-67,p53 and nm23 were analyzed.Results The expression level of CCDC8 in benign breast tissue (1685±755) was significantly higher than that in cancerous tissues (502.1 ±223.2).Tissues obtained from patients over age 50 showed an increased level of CCDC8 (789.8±367) in comparison to those from patients age 50 or younger (452.5±170.3).The level of CCDC8 expression was negatively correlated with nm23 level (Correlation Coefficient =-0.400,P =0.039),while no correlation was found between CCDC8 and cancer stages,estrogen and progesterone receptor,CerbB2,Ki-67and p53.Conclusion The negative correlations between CCDC8 and age,tumor size and nm23 indicate that CCDC8 is a potential tumor suppressor,influencing the occurrence and progression in breast cancer.

Radiotherapy plays an important role in the current treatment of breast cancer, which can greatly reduce the local recurrence rate of breast cancer patients. In recent years, many clinical trials on the radiotherapy of breast cancer have shown that the benefits of radiotherapy are different in breast cancer patients with different molecular typing, lymph node metastasis, surgical resection methods, and so on. Therefore, in the era of precision medicine, the radiotherapy of breast cancer needs to be based on the clinical treatment guidelines and clinical evidence, and in combined with the actual situation. The precise radiotherapy should be carried out according to the assessment results of individual recurrence risk, so as to maximize the benefit of patients through reducing the recurrence rate and avoiding excessive treatment. In this paper, the current research progress in the post-operative radiotherapy based on the molecular classification of breast cancer, lymph node metastasis and surgical resection methods is summarized, in order to provide the clinical basis of postoperative precise radiotherapy for breast cancer patients.

Objective To evaluate the effect of photoaging on the degradation of advanced glycation end products (AGEs) by human dermal fibroblasts.Methods Some cultured human dermal fibroblasts were subjected to repetitive ultraviolet A (UVA) radiation (UVA radiation group) to establish a photoaging cell model,which was then evaluated by cell counting kit 8 (CCK-8) assay,senescenceassociated β-galactosidase staining and detection of apoptosis rate.Moreover,fibroblasts receiving no treatment served as control group.Some other primary fibroblasts were divided into 4 groups:photoaged group receiving UVA radiation,non-photoaged group receiving no treatment,AGE-treated photoaged group treated with UVA radiation followed by the treatment with 200 mg/L AGE-bovine serum albumin (BSA),and AGE-treated non-photoaged group treated with 200 mg/L AGE-BSA alone.After the treatment with AGE-BSA for 4-72 hours,flow cytometry was performed to determine the fluorescence intensity of AGE-BSA in fibroblasts of the above groups.After 8-hour treatment with AGE-BSA,confocal laser scanning microscopy was performed to localize and semiquantitatively detect AGE-BSA in fibroblasts,and enzymelinked immunosorbent assay (ELISA) was conducted to detect AGE-BSA levels in fibroblasts,as well as changes in the intracellular AGE-BSA level within 24 hours after the removal of AGE-BSA.Results Compared with the control group,the UVA radiation group showed significantly decreased cellular proliferative activity (t =7.559,P ＜ 0.05),but significantly increased apoptosis rate and percentage of β-galactosidase-positive fibroblasts (t =14.075,43.524 respectively,both P ＜ 0.05).Flow cytometry revealed that the average fluorescence intensities of AGE-BSA after 4-,8-,16-,24-,48-and 72-hour treatment with AGE-BSA were significantly higher in the AGE-treated photoaged group (293.00 ± 8.19,359.67 ± 11.59,347.00 ± 12.29,338.00 ± 12.77,334.67 ± 14.22 and 336.30 ± 10.21,respectively) than in the photoaged group (all P ＜ 0.05),as well as in the AGE-treated non-photoaged group (222.33 ± 8.74,276.33 ± 6.11,256.33 ± 5.51,243.00 ± 10.15,236.33 ± 1.53 and 240.33 ± 1.52,respectively) than in the non-photoaged group (all P ＜ 0.05).Moreover,the average fluorescence intensities of AGE-BSA at different time points were all significantly higher in the AGE-treated photoaged group than in the AGE-treated non-photoaged group (all P ＜ 0.05).Confocal laser scanning microscopy showed that AGE-BSA was mainly localized in lysosomes after endocytic uptake into the fibroblasts,and the AGE-treated photoaged group showed significantly increased fluorescence intensity of AGE-BSA compared with the AGE-treated non-photoaged group (P ＜ 0.05).ELISA revealed that the intracellular AGE level in the AGE-treated non-photoaged group at 24 hours after the removal of AGE-BSA was decreased by (14.6 ± 1.2)％ compared with that before the removal,and the degradation rate of AGE-BSA was significantly higher in the AGE-treated non-photoaged group than in the AGE-treated photoaged group (7.6％ ± 1.4％,t =6.604,P ＜ 0.05).Conclusion The internalized AGE-degradating ability decreases in photoaged fibroblasts,which may induce the accumulation of AGEs in photoaged skin.

Objective To determine the expression of cathepsin D and advanced glycation end products (AGEs)in skin tissues from patients of different ages or skin tissues with different degrees of sun exposure,to evaluate their correlation,and to preliminarily investigate the role of cathepsin D in the degradation and accumulation of AGEs in photoaged skin.Methods Skin tissues were collected from sunexposed and sun-protected body sites in patients aged 15-20 years,35-40 years,55-60 years or 75-80 years.These skin tissues were divided into 8 groups according to age of patients and degrees of sun exposure,and there were 6 specimens in each group.Immunohistochemical and immunofluorescent methods were used to measure the expression of cathepsin D and AGEs in the skin tissues.Statistical analysis was carried out by factorial design analysis of variance,Wilcoxon rank sum test and Kruskal-Wallis rank sum test for analyzing associations of the expression of cathepsin D and AGEs with age and sun exposure,as well as by Pearson correlation analysis for assessing the correlation between cathepsin D expression and AGEs expression.Results Immunohistochemical study showed that the expression of cathepsin D markedly decreased along with the increase of age,but the accumulation of AGEs gradually increased along with the increase of age.In the same age group,the cathepsin D expression was lower in the sun-exposed skin tissues than in the sun-protected skin tissues,while the accumulation of AGEs was more in the sun-exposed skin tissues than in the sun-protected skin tissues.Factorial design analysis of variance showed that sun exposure could decrease the expression of cathepsin D (F =58.70,P ＜ 0.001),but increase the accumulation of AGEs (F =158.18,P ＜ 0.001).Moreover,the increase of age could lead to decreased expression of cathepsin D (F =79.49,P ＜ 0.001),and increased expression of AGEs (F =106.06,P ＜0.001).Compared with the sun-protected skin tissues,the sun-exposed skin tissues in all the age groups showed significantly lower absorbance value of cathepsin D (35-40 years:0.020 ± 0.005 vs.0.032 ± 0.005;55-60 years:0.012 ± 0.004 vs.0.026 ± 0.002;75-80 years:0.002 ± 0.001 vs.0.013 ± 0.004;all P ＜0.001),but higher absorbance value of AGEs (35-40 years:0.030 ± 0.008 vs.0.010 ± 0.003;55-60years:0.066 ± 0.010 vs.0.021 ± 0.004;75-80 years:0.085 ± 0.015 vs.0.035 ± 0.009;all P ＜ 0.001)except the age group of 15-20 years.No matter whether the skin tissues were sun-exposed or sunprotected,there were significant differences in the expression of cathepsin D and AGEs among different age groups (all P ＜ 0.001).The results of double immunofluorescence staining were similar to those of immunohistochemical study.Pearson correlation analysis showed that the expression of cathepsin D in the sun-exposed skin tissues was highly negatively correlated with the accumulation of AGEs (r =-0.915,P ＜0.05),while they were moderately negatively correlated in the sun-protected skin tissues (r =-0.730,P ＜0.05).Conclusions Along with the increase of age,the expression of cathepsin D in skin tissues decreased,but the expression of AGEs increased.In the sun-protected skin tissues,the expression of cathepsin D was moderately negatively correlated with the expression of AGEs,while they were highly negatively correlated in the sun-exposed skin tissues,suggesting that cathepsin D may play an important role in the degradation and accumulation of AGEs in photoaged skin.

Objective To investigate the regulatory role of cathepsin D (CatD) in the degradation of intracellular advanced glycation end products (AGEs) endocytosed by human dermal fibroblasts (HDFs).Methods Cultured HDFs were treated with 1 μnol/L CA074Me (an inhibitor of CatB and CatL),75 μmol/L pepstatin A (an inhibitor of CatD) and 1 μmol/L MG-132 (an inhibitor of20S proteasome) separately for 4 hours,and then cell counting kit 8 (CCKS) assay and fluorometric assay were performed to determine the cellular viability and protease activity,respectively.The cells in the CA074Me group,pepstatin A group and MG-132 group were additionally treated with AGE-bovine serum albumin (BSA) for 8 hours,and the cells in the blank control group were treated with phosphate-buffered saline (PBS) alone.After 8-hour cultivation,the cells in the above groups were subsequently reincubated with fresh culture medium containing the corresponding inhibitors for 24 hours.Then,flow cytometry was performed to assess the mean fluorescence intensity of intracellular AGE-BSA at different time points.Some other HDFs were treated with 37.5,75 and 150 μmol/L pepstatin A and PBS separately for 4 hours,and then the cells in the 4 groups were treated with 200 mg/L AGE-BSA for 8 hours,followed by the removal of AGE-BSA from the medium and the treatment with 37.5,75 and 150 μmol/L pepstatin A and PBS respectively.Enzyme-linked immunosorbent assay (ELISA) was conducted to measure the mean concentration of intracellular AGE-BSA at different time points,and the degradation rate of AGE was calculated.Some HDFs were divided into 3 groups:blank control group receiving no treatment,NC group transfected with an empty vector,and CatD group transfected with a CatD-overexpressing lentiviral vector.Fluorescence microscopy was conducted to estimate the transfection efficiency.Reverse transcription-PCR,Western blot analysis and fluorometric assay were performed to determine the mRNA and protein expression,and activity of CatD respectively.Then,the cells in the above 3 groups were incubated with AGE-BSA for 8 hours,followed by the removal of AGE-BSA from the medium and the treatment with fresh culture medium.The detection methods were same as the above experiment,and the degradation rate was calculated.Results The cellular proliferative activity in the 1-μmol/L CA074Me group,75-μmnol/L pepstatin A group and 1-μ mol/L MG-132 group was more than 90％,and there was no significant difference between the 3 groups and the control group (100％,F =1.525,P ＞ 0.05).Twenty-four hours after the removal of AGE-BSA from the medium,the fluorescence intensities of intracellular AGE-BSA in the CA074Me + AGE-BSA group (275.00 ± 10.15) and MG-132 + AGE-BSA group (259.00 ± 11.14) significantly decreased compared with those at the 8-hour time point (295.00 ± 6.56 and 285.67±8.74 respectively;paired t test,t =4.778,6.154 respectively,both P ＜ 0.05),while no significant difference was observed in the fluorescence intensities of intracellular AGE-BSA in the pepstatin A + AGE-BSA group between the 8-hour time point and 32-hour time point (P ＞ 0.05).The degradation rates of intracellular AGE-BSA within 24 hours in the 37.5,75 and 150 μmol/L pepstatin A groups were 9.64％ ± 1.27％,5.62％ ± 0.47％ and 3.21％ ± 0.73％ respectively;there were significant differences among the 3 groups (F =45.876,P ＜ 0.05),and the degradation rate significantly decreased along with the increase of pepstatin A concentration (P ＜ 0.05).Fluorescence microscopy showed no fluorescent cells in the blank control group,while the NC group and CatD group both showed a high proportion (＞ 80％) of fluorescent cells.The mRNA and protein expression as well as the activity of CatD were significantly higher in the CatD group than in the blank control group and NC group (all P ＜ 0.05).The CatD + AGE-BSA group showed a significantly higher degradation rate of intracellular AGE-BSA within 24 hours compared with the AGE-BSA group and NC + AGE-BSA group (both P ＜ 0.05).Conclusion CatD can promote the degradation of intracellular AGE-BSA endocytosed by HDFs.

【Objective】 To determine the best collection time period of plasma which can be used for human COVID-19 immunoglobulin for intravenous injection through SARS-CoV-2-IgG change and neutralizing antibody distribution against different virus strain in representative mixed plasma before and after Omicron strain infection by ELISA and pseudovirus neutralization test. 【Methods】 An ELISA method for quantitative detection of SARS-CoV-2-IgG was established and its linear range,accuracy and precision was verified. SARS-CoV-2-IgG potency was detected in 25 convalescent plasma which were collected 20-40 days after confirmed Omicron infection, two groups of mixed plasma samples WP1 and WP2 were prepared according to the SARS-CoV-2-IgG results, and pseudovirus neutralization experiments with different virus strain (prototype strain, BA. 1,BA.2, BA.4/5, BF.7, BQ.1.1) were carried out to determine the distribution of neutralizing antibodies against different virus strain. SARS-CoV-2-IgG potency of representative mixed plasma collected from 14 plasma stations subordinate to the company before and after Omicron strain infection was detected, including Omicron convalescent plasma (OP) collected from different plasma stations from December 2022 to May 2023 and normal pool plasma (VN) feed in March 2023 which collected from March 2022 to December 2022. According to the results, the difference and the change rule with time of SARS-CoV-2-IgG before and after Omicron strain infection were analyzed. 【Results】 The linearity of SARS-CoV-2-IgG ranged from 6.25 to 200 EIU/mL, the accuracy in-batch ranged from 81.793% to 106.985%, the precision in-batch ranged from 1. 100% to 13.000%, and the total error in-batch ranged from 2.988% to 22.679%. The accuracy between batches ranged from 90.788%to 96.893%, the precision between batches ranged from 4.870% to 6.272%, and the total error between batches ranged from 9.192% to 15.399%. The results of pseudovirus neutralizing antibody showed that the potency of different virus strain neutralizing antibodies were in the order of prototype strain>BA.2>BA.4/5>BF.7≈ BQ.1.1>BA.1 and the correlation between WP1 and WP2 was high (Pearson r=0. 931 1, P=0.002 3) which indicated that the potency distribution of neutralizing antibodies of different virus strain in Omicron convalescent plasma was basically stable. Compared with the mixed convalescent plasma sample G128 collected in June 2022, the potency of Omicron neutralizing antibodies of WP series were significantly higher, the ratio of BA.2 antibody to prototype antibody increased from 26.9% (before infection) to 82.6%-87.5% (after infection). The results of VN series before Omicron infection were < 100 EIU/mL, and the results of OP series after Omicron infection showed that the plasma collected from the beginning of December 2022 was the peak of antibody in the same month,and then dropped sharply, entering a short plateau in February-March 2023 (potency was about 40% of the peak value),and then dropped sharply again in April (potency was about 20% of the peak value). 【Conclusion】 The potency and proportion of neutralizing antibody against Omicron subtype in convalescent plasma after COVID-19 Omicron strain infection increased significantly. IgG antibody of plasma donors in different regions reached its peak in the month of infection, then continued to dropped sharply. The best collection period of plasma that can be used for human COVID-19 immunoglobulin for intravenous injection was 1 to 2 months after infection.

OBJECTIVE To study the extraction and enrichment technology of chrysosplenides A （CA） and I （CI） in Tibetan medicine Chrysosplenium axillare. METHODS HPLC method was used to determine the contents of CA and CI. The orthogonal experiment was used to optimize the extraction technology of CA and CI in C. axillare using total transfer rate of CA and CI as evaluation indexes， with volume fraction of ethanol， extraction temperature， extraction times and solid-liquid ratio as factors. The validation test was also performed. The enrichment technology of CA and CI in C. axillare was optimized using D101 macroporous adsorption resin as adsorbent， total contents of CA and CI as evaluation indexes， with the volume fraction and dosage of eluent for impurities and target components. The validation test was also performed. RESULTS The optimum extraction conditions of CA and CI from C. axillare were as follows： the medicinal powder of C. axillare was extracted by ultrasound at room temperature for 45 min at one time with 8 times of 50% ethanol. Results of validation tests showed that total transfer rate of CA and CI in C. axillare was 95.43% in average （RSD＝1.02%， n＝3）. The optimal enrichment technology was as follows： the sample solution was added into D101 macroporous adsorption resin column and stood for 1 hour； the impurities were eluted with 20% ethanol 4 BV （column volume）， and CA and CI were eluted with 50% ethanol 4 BV. The results of validation tests showed that total content of CA and CI was 322.7 mg/g in average （RSD＝1.05%， n＝3）， with average enrichment multiple of 11.61 times. CONCLUSIONS The study has successfully optimized the extraction and enrichment technology of CA and CI from C. axillare， and can provide reference for the development and utilization of CA and CI.