Objective To compare the efficacy between ureteroscope with holmium laser lithotripsy (URS) and minimally invasive percutaneous nephrolithotomy (MPCNL) in oblique supine position for the proximal and middle ureteral calculi.Methods Ninety-six patients with proximal and middle ureteral calculi were divided into URS group (54 cases) and MPCNL group(42 cases) according to the treatment order.The operative duration,postoperative hospital stay,stone-free rate,intraoperative blood loss ＞ 400 ml rate,conversion to open surgery rate,complication rate including postoperative fever were observed,and stone composition was analyzed.Results The operative duration and postoperative hospital stay in URS group and MPCNL group were (55.3 ± 12.1),(49.6 ± 11.5) min and (1.7 ± 0.9),(4.4 ± 1.2) d,respectively,and there were significant differences (P ＜ 0.05 or ＜ 0.01).There were no significant differences in the stone-free rate,complication rate (Clavien degree Ⅱ),conversion to open surgery rate,intraoperative blood loss ＞ 400 ml rate,complication rate,postoperative fever rate (P ＞ 0.05).Stone composition analysis showed that the proportion of calcium oxalate stone was 51.0％(49/96).Conclusions URS and MPCNL in oblique supine position for the proximal and middle ureteral calculi are effective and safe therapeutic modalities.The operative duration of MPCNL in oblique supine position is shorter,and postoperative recovery of URS is fast.

Objective The effect of fertilizer application on yield and quality of Notopterygium forbesii was studied in order to provide a theoretical basis for high quality and yield planting.Methods With various nitrogenous(N) and phosphorous(P) fertilizer formula,the field tests were carried out to determine the yield,extract content,and essential oil content,from the root of N.forbesill.Results Showed that various proportion of fertilizer had a great affect on yield,extract content,and essential oil content,and the treatment with N2P2 was the highest.The yield in the treatment with N2P2 increased 43.33%,the extract content increased 17.95%,and the essential oil content increased 43.48% as compared with CK.Different rates of fertilizer application had a certain effect on weight and diameter in the root of N.forbesii,and the N2P2 was the best.Each treatment had a significant effect on diameter in the root of N.forbesii in comparision with CK.Conclusion Amount and ratio of N and P fertilizers have a greater influence on the yield and quality,while improper application could reduce the yield and quality.

Objective To knock out Asxl2 gene in murine embryonic fibroblast cell line NIH3T3 using CRISPR/Cas9n system. Methods A pair of sgRNAs which targeted exon 5 of Asxl2 gene were designed and subcloned into the pX462 vec?tor. The recombined plasmids were verified by sequencing and transfected into NIH3T3 cell line. Single cells were isolated through serial dilutions, followed by an expansion period to obtain new monoclonal cell lines. The genomic DNA of the new monoclonal cell lines was extracted and a DNA fragment flanked the target site was amplified by genotyping PCR then se?quenced. Lastly, western blotting were applied to confirm whether Asxl2 was successfully knocked out. Results The CRIS?PR/Cas9n plasmids that targeted Asxl2 were successfully constructed. NIH3T3 cells were co-transfected with the two recom?binant constructs. After puromycin selection, subclonal cell lines were obtained and one of them was validated by genotyping PCR-sequencing. Western blotting also confirmed that Asxl2 was completely depleted in the NIH3T3 cell line. Conclu?sion CRISPR/Cas9n plasmids that targeted Asxl2 were successfully constructed therefore a Asxl2 knockout NIH3T3 stable cell line was established via this system.

Objective To knock out p21 gene in human malignant rhab doid tumor(MRT)cell line G401 by using CRISPR/Cas9 genome engineering technology. Methods The expression of p21 was detected by reverse transcription quantitative PCR (RT-qPCR) and Western blot assay in several MRT cell lines. The guide RNA was designed by targeting the third exon of p21 gene,which encoded its home domains, and then subcloned into lentiCRISPR v2 vector and validated sequencing. The validated plasmids were further used to package and produce the lentivirus in 293T cells, and the G401 cells were infected, then puromycin was used to screen positive cells, and the clusters of G401 monoclonal cells, were obtained by selecting monoclonal cells and culturing under the microscope. The RNA and protein of new clonal cell line were extracted, and RT-qPCR and Western blot assay were applied to confirm whether p21 was successfully knocked out. Results The p21 was highly expressed in MRT tumor cells. The CRISPR/Cas9 lentivirus plasmids, targeted p21 gene were successfully constructed. Compared with negative control group,the expression of p21 was not detected in G401 monoclonal cells, which were successfully screened. Conclusion In view of the difficult transfection of cells such as G401, p21 knockout stable cell line has been successfully constructed by using CRISPR/Cas9 system, which lays the foundation for further study of the mechanism of p21 in MRT tumors .

Objective To predict the potential suitable growth areas of Epimedium brevicornu Maxim.under current climate conditions in China;To provide basis for the sustainable utilization of Epimedium brevicornu Maxim.resources and production planning.Methods Based on 267 screened species distribution data of Epimedium brevicornu Maxim.and 8 environmental factors,the MaxEnt parameters were optimized by the R language kuenm package.The main environmental factors that affect the distribution of Epimedium brevicornu Maxim.was analyzed and its potential distribution range was predicted.Results The omission rate of the optimal model operation results was 0.044 8,AICc=6 409.884 5,AUC=0.986,indicating a high accuracy of the model.Based on the contribution rate of environmental factors and the knife cut method,it was indicated that the key environmental factors affecting the current distribution of Epimedium brevicornu Maxim.were mainly the average precipitation in July,the average highest temperature in February,the average precipitation in September,the coldest season precipitation,the coefficient of variation of precipitation,the standard deviation of seasonal temperature changes,and the warmest season precipitation.The simulation results showed that the total suitable area for Epimedium brevicornu Maxim.in China under current climate conditions was approximately 221.14×104 km2,concentrated in the central and central southern regions of China.The area of the high suitability zone was approximately 23.13×104 km2;the area of the suitable growth zone was approximately 73.78×104 km2;the area of low suitability zone was approximately 124.22×104 km2.Conclusion This study provides a basis and reference for the artificial cultivation planning and sustainable utilization of Epimedium brevicornu Maxim.resources.

Objective: To study the survival time and its risk factors of patients with occupational pneumoconiosis. Methods: A total of 11 011 newly diagnosed occupational pneumoconiosis patients in Guangdong Province from 1980 to 2019 were selected as study subjects. The life table method was used for survival analysis. The influencing factors of survival time of occupational pneumoconiosis patients were analyzed using the WilCoxon (Gehan) test and Cox proportional hazards regression model. Results: The median survival time of pneumoconiosis patients was 26.0 years. The median survival period of stage Ⅰpatients was 3.5 years longer than that of stage Ⅱ patients and 10.1 years longer than that of stage Ⅲ patients. The median survival time of patients with an initial diagnosis age under 40.0 years old was 34.8 years longer than that of patients with an initial diagnosis age over 60.0 years old. The median survival time of patients with dust exposure duration under 25.0 years old was 13.6 years longer than patients with dust exposure duration age over 45.0 years old. The results of the Cox proportional hazards regression model showed that the initial diagnosis stage, initial diagnosis age, dust exposure duration, and medical insurance were risk factors of the survival time of occupational pneumoconiosis patients (all P<0.01). The risk of reduced survival time for patients with stage Ⅱ and stage Ⅲ as the initial diagnosis stage was 1.15 and 2.04 times higher, respectively, compared with stage Ⅰ patients (both P<0.01). The risk of reduced survival time for patients without medical insurance was 60.22 times higher than those with medical insurance (P<0.01). Conclusion: The risk factors of the survival time of occupational pneumoconiosis patients in Guangdong Province are initial diagnosis stage, initial diagnosis age, the dust exposure age, and medical insurance. Earlier detection, earlier diagnosis, and improvement of medical insurance coverage for patients can effectively improve the survival time of occupational pneumoconiosis patients.

Objective: To verify the accuracy of the autoregressive integrated moving average (ARIMA) in predicting the incidence of occupational pneumoconiosis (hereinafter referred as pneumoconiosis) and to predict the incidence of pneumoconiosis in Guangdong Province in the next five years. Methods: A follow-up survey was performed to collect data on pneumoconiosis patients reported in Guangdong Province from 1956 to 2021. Collected data from 1956 to 2016 were used as the training set to build an ARIMA model. Collected data from 2017 to 2021 were used as the prediction set to evaluate the predicting result of the ARIMA model. The ARIMA model was used to predict the incidence of pneumoconiosis in Guangdong Province in next five years. Results: The ARIMA (1,1,2) model was set up after model identification and order estimation. The model was used to predict the prediction set, and its result was good. The ARIMA result and actual values in 2021 were 213 and 210 cases, respectively, with a difference of only three cases. The number of pneumoconiosis cases predicted using the ARIMA model in Guangdong Province from 2022 to 2026 was 214, 204, 202, 194, and 191 cases, respectively, showing a trend of low-level prevalence. Conclusion: The ARIMA model demonstrates high accuracy in predicting pneumoconiosis incidence over a long period of time and with large sample sizes. The forecast results of the ARIMA(1,1,2) model indicate that the incidence of pneumoconiosis in Guangdong Province will be around 200 cases in the next five years, indicating a low-level prevalence.

By consulting the ancient and modern literature， the textual research of Pharbitidis Semen has been conducted to clarify the name， origin， distribution of production areas， quality specification， harvesting， processing and so on， so as to provide reference for the development and utilization of the relevant famous classical formulas. Through textual research， it can be seen that Pharbitidis Semen was first published in Mingyi Bielu（《名医别录》）， and all dynasties have taken Qianniuzi as the correct name. Based on the original research， the main source of Pharbitidis Semen used in previous dynasties is the dried mature seeds of Pharbitis nil， which is consistent in ancient and modern times. The white Pharbitidis Semen appearing in Compendium of Materia Medica（《本草纲目》） from Ming dynasty is similar to the present P. purpurea. It is produced all over the country， and the quality is better if the particles are full and free of impurities. In ancient times， the harvesting time was mostly in the September. Now it is autumn. The fruits are ripe and harvested， dried to remove impurities for standby. In ancient times， the processing methods of Pharbitidis Semen were mainly wine steaming， steaming and frying until half cooked and grinding the head and end. In modern times， they have been simplified to stir-frying method. The nature， taste， meridian tropism and their effects also change supplements with the deepening of practice. Before the Ming dynasty， they were all bitter， cold and toxic. In the Ming dynasty， there appeared the characteristics of pungent， hot and small poisonous. The efficacy has evolved from controlling low Qi， curing foot edema， removing wind toxin， and facilitating urination to facilitating water and defecation， eliminating phlegm and drinking， and eliminating accumulated insects. The main clinical contraindications are those with weak spleen and kidney， those with weak spleen and stomach， pregnant women， and should not be used with croton and croton cream. Based on the textual research， it is suggested that when developing the classic famous formula with Pharbitidis Semen as the main raw material in the future， it is clear that the source should be the dried mature seeds of Pharbitis nil（black product is its black-brown seeds， white product is its beige seeds）. The processing requirements indicated in the original formula are all processed according to the requirements， and the raw product is recommended to be used as medicine if not specified.