1.Effects of different storage temperatures and durations on the activity of coagulation factor Ⅷ and Ⅸ in whole blood
Hehe WANG ; Tiantian WANG ; Jie WANG ; Cuicui QIAO ; Wei LIU ; Xueqin ZHANG ; Yan CHENG ; Yunhai FANG ; Xinsheng ZHANG
Chinese Journal of Blood Transfusion 2025;38(6):824-827
		                        		
		                        			
		                        			Objective: To investigate the effects of different storage temperatures and durations on the activities of coagulation factor Ⅷ (Factor Ⅷ, FⅧ) and coagulation factor Ⅸ (Factor Ⅸ, FⅨ) after whole blood collection, so as to provide data support for the optimal storage conditions. Methods: A total of 16 mL of whole blood was collected from each of the 20 healthy volunteers at our blood center and aliquoted into 8 sodium citrate anticoagulant tubes. Two tubes were immediately centrifuged for the measurement of FⅧ and FⅨ activity levels. The remaining 6 tubes of whole blood were respectively stored under room temperature and low-temperature conditions. At 2, 4, and 6 h, the whole blood samples were centrifuged and analyzed for FⅧ and FⅨ activity levels. The mean values of the two immediately tested tubes were used as the control group, while other tubes were designated as the experimental groups for comparison. Statistical analysis was performed using SPSS 26.0. Results: The activity of FⅧ in whole blood remained stable after 4 hours of storage at both room temperature and low temperature (116.53±25.95 vs 125.22±27.33, 109.77±23.23 vs 125.22±27.33) (P>0.05 for both). However, by 6 hours, FⅧ activity showed a statistically significant decline compared to the control group (108.65±22.92 vs 125.22±27.33, 100.46±20.19 vs 125.22±27.33) (P<0.05 for both), though the room temperature group results were closer to the control values. The activity of FⅨ in whole blood remained stable after 6 hours of storage under both conditions (97.14±19.48 vs 96.76±19.67, 97.10±17.45 vs 96.76±19.6) (P>0.05 for all comparisons). Conclusion: For whole blood samples after collection, storage at either room temperature or low temperature for up to 4 hours does not compromise the accuracy of test results. When stored for 6 hours, FⅨ activity remains stable, whereas FⅧ activity decreases significantly. Notably, FⅧ activity demonstrates better stability at room temperature than under low-temperature conditions within the 6-hour storage.
		                        		
		                        		
		                        		
		                        	
2.Effects of prescription pre-review system on rational drug use and off-label drug use management in outpatient and emergency department
Zhi GAO ; Lulu HAN ; Fang LIU ; Rui JIAO ; Wei ZHANG ; Yi ZHANG
China Pharmacy 2025;36(13):1666-1670
		                        		
		                        			
		                        			OBJECTIVE To explore the effects of prescription pre-review system on rational drug use and off-label drug use management in outpatient and emergency department. METHODS A retrospective analysis was conducted on outpatient and emergency department prescription data from three phases in our hospital: January to May 2023 (silent review phase, control group), June to October 2023 (systematic automatic review phase, intervention group 1), and November 2023 to March 2024 (phase combining systematic automatic review with centralized feedback from pharmacists to physicians regarding irrational prescriptions, intervention group 2). These phases followed the implementation of our hospital’s pre-prescription review software. Statistical analysis of the prompt rate of alert, rate of irrational prescriptions, registered the off-label drug use rate and false positive irrationality prescription rate were conducted. Meanwhile, the composition of irrational prescriptions was analyzed, and evidence- based evaluation of the off-label drug use proposed by clinicians was also conducted. RESULTS Compared with control group, the prompt rate of alert and the rate of irrational prescriptions in intervention group 1 were all decreased significantly after receiving pop-up notification, with statistically significant differences (P<0.05). With the help of system warning and the pharmacists feedback, the prompt rate of alert and the rate of irrational prescriptions declined further in the intervention group 2, but there was no statistically significant difference when compared with intervention group 1 (P>0.05). The main type of irrational drug use was improper administration routes. When comparing intervention group 1 with the control group, as well as intervention group 2 with intervention group 1, a significant decrease in the rate of improper administration routes was observed, with statistically significant differences (P<0.05). Compared with control group, there was no significant difference in the registered off-label drug use rate of intervention group 1 and intervention group 2 (P>0.05). The doctor’s awareness of off-label drug use registration increased due to the real-time alerts from the pre-prescription review software, along with the pharmacists’ regular summarization and feedback. Total 13 items registrations of off-label drug use were proposed by clinicians from June 2023 to March 2024, all of which were supported by evidence of varying levels; among them, 3 items received FDA approval, 4 items were included in the Micromedex database, and the remaining 6 items were supported by evidence from system reviews or randomized controlled trials. CONCLUSIONS Prescription pre-review system can improve the level of rational drug use and assist in the standardized management of off-label drug use.
		                        		
		                        		
		                        		
		                        	
3.Quality evaluation of Sanzi powder based on quantitative analysis of multi-component combined with chemical pattern recognition and entropy weight-TOPSIS method
Rongjie LI ; Qian ZHANG ; Wei ZHANG ; Xinkui LI ; Yuxia HU ; Mengdi ZHANG ; Jing LIU ; Fang WANG ; Fengye ZHOU ; Jun LI
China Pharmacy 2025;36(15):1846-1851
		                        		
		                        			
		                        			OBJECTIVE To comprehensively evaluate the quality of Sanzi powder from different batches based on 12 components quantitative analysis combined with chemical pattern recognition and entropy weight-TOPSIS method. METHODS The contents of 12 components in 15 batches of Sanzi powder (No. S1-S15) were determined by HPLC-MS/MS, such as ethyl gallate, gallic acid, ferulic acid, corilagin, genipin-1-O-β-D-gentiobioside, toosendanin, geniposide, caffeic acid, methyl deacetylated coumarinate, tannic acid, rutin, quercetin. Cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were conducted on the assay results. Using variable importance projection (VIP) value>1 and P<0.05 as the evaluation criteria, the quality differential markers in Sanzi powder were screened. The entropy weight method was used to calculate the weight value, and TOPSIS method was used to rank the quality of 15 batches of Sanzi powder from superior to inferior. RESULTS The contents of the 12 components were 13.494-24.292, 2 069.608-3 188.100, 1.410-3.616, 1 065.030-2 630.584, 1 404.704-1 838.078, 101.640-354.268, 9 193.720-14 777.854, 1.240-5.060, 148.028-5 541.990, 4 261.422-5 607.438, 107.560- 195.512, 2.226-4.192 μg/g, respectively. The results of CA, PCA and OPLS-DA indicated that 15 batches of Sanzi powder could be clustered into two groups. Specifically, batches S3, S7, S10 and S15 were grouped into one category, and remaining batches were grouped into one category. VIP values of geniposide, quercetin, caffeic acid, and methyl deacetylated coumarinate were all greater than 1, with corresponding P-values less than 0.05. The results of the entropy weight-TOPSIS analysis revealed that methyl deacetylate exhibited the smallest information entropy and the highest weight. The relative closeness degrees of samples S3, S7, S10 and S15 ranged from 0.789 to 0.973, while the remaining samples ranged from 0.054 to 0.172. CONCLUSIONS The contents of 12 components in Sanzi powder could be determined accurately by using HPLC-MS/MS technology. Methyl deacetylated coumarinate, geniposide, quercetin and caffeic acid were identified as the quality differential markers. It was found that the overall quality of samples S3, S7, S10 and S15 were superior to that of other batches. Notably, the quality of Gardeniae Fructus decoction pieces emerges as a critical factor in ensuring the consistency of the preparation’s quality.
		                        		
		                        		
		                        		
		                        	
4.Mechanism of Yantiao Prescription in Treating Lipopolysaccharide-induced Acute Lung Injury Based on Arachidonic Acid Metabolic Pathways
Pengcheng LI ; Tianyang CHEN ; Rong FANG ; Anna ZHANG ; Sijia WU ; Wei LIU ; Qian WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):101-110
		                        		
		                        			
		                        			ObjectiveTo clarify the anti-inflammatory and lung-protective effects of Yantiao prescription on lipopolysaccharide (LPS)-induced acute lung injury (ALI), and to explore the impact of Yantiao prescription on the metabolic pathways of arachidonic acid (AA) in vivo. MethodsThirty male C57BL/6J mice were randomly divided into the following groups based on body weight: normal group, model group, dexamethasone group (2 mg·kg-1), low-dose Yantiao prescription group (18 g·kg-1), and high-dose Yantiao prescription group (36 g·kg-1), with 6 mice in each group. The ALI mouse model was established by intraperitoneal injection of LPS. The treatment groups received oral gavage once a day for 7 consecutive days, and serum and lung tissue were collected at the end of the experiment. The content of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in serum was detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was used to assess lung tissue pathology. The wet/dry weight ratio (W/D) and myeloperoxidase (MPO) activity in lung tissue were measured. The content of AA metabolites in serum and lung tissue was measured by liquid chromatography triple quadrupole-mass spectrometry (LC-MS/MS). ResultsCompared with the conditions in the normal group, the content of serum pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in the model group was significantly increased (P<0.01). The alveolar structure in mice was severely damaged, with markedly thickened alveolar walls and extensive inflammatory cell infiltration. The W/D ratio and MPO activity in lung tissue were significantly increased (P<0.01). The content of AA metabolites, including prostaglandin D2 (PGD2), prostaglandin E2 (PGE2), 11(S)-hydroxy-eicosatetraenoic acid [11(S)-HETE], and 5-hydroxy-eicosatetraenoic acid (5-HETE) in serum and lung tissue was significantly increased (P<0.05), while the content of 11,12-epoxyeicosatrienoic acid (11,12-EET) and 14,15-epoxyeicosatrienoic acid (14,15-EET) in serum was significantly decreased (P<0.01). Compared with the results in the model group, the content of serum pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in the dexamethasone group, low-dose Yantiao prescription group, and high-dose Yantiao prescription group was significantly reduced (P<0.05). Mild thickening of alveolar walls, scattered inflammatory cell infiltration, and relatively intact tissue structure with improved alveolar architecture were observed. The W/D ratio and MPO activity in lung tissue were significantly reduced (P<0.01). The content of AA metabolites PGD2, PGE2, 11(S)-HETE, and 5-HETE in serum from the dexamethasone group was significantly decreased (P<0.05), while the content of 14,15-EET in serum significantly increased (P<0.01), and the content of 5-HETE in lung tissue significantly decreased (P<0.01). In the low-dose and high-dose Yantiao prescription groups, the content of AA metabolites PGD2, PGE2, 11(S)-HETE, and 5-HETE in serum and lung tissue was significantly decreased (P<0.05), while the content of 11,12-EET in both serum and lung tissue was significantly increased (P<0.05). ConclusionYantiao prescription has significant protective effects against LPS-induced ALI, which are related to its regulation of AA metabolic pathways in vivo. 
		                        		
		                        		
		                        		
		                        	
5.Prognostic Factors of Liposarcoma in Head and Neck
Shuo DING ; Zhigang HUANG ; Jugao FANG ; Yang ZHANG ; Lizhen HOU ; Wei GUO ; Gaofei YIN ; Qi ZHONG
Cancer Research on Prevention and Treatment 2025;52(1):31-35
		                        		
		                        			
		                        			Objective To explore the pathogenesis and prognostic factors of liposarcoma in the head and neck region, and simultaneously analyze the efficacy of different treatment regimens. Methods A retrospective analysis was performed on all patients with primary untreated head and neck liposarcoma who were diagnosed and underwent surgical treatment at our hospital from January 2008 to January 2024. All patients were monitored during follow-up, and their prognoses were analyzed using SPSS software. Results A total of 30 patients were included in the study. Liposarcoma accounted for up to 60% of the cases in the orbit, while the remaining liposarcomas were primarily located in various interspaces of the neck. Dedifferentiated liposarcoma was the most common type, comprising 33%, while myxoid pleomorphic liposarcoma was the rarest at 4%. The tumor pathological type (P<0.001) and Ki67 (P=0.014) significantly affected the tumor control rate. However, an analysis of disease-specific survival rates revealed no significant differences across various factors (all P>0.05). Conclusion The prognosis of head and neck liposarcoma is better compared to that of liposarcomas in other parts of the body. However, myxoid pleomorphic liposarcoma, pleomorphic fat sarcoma, and high Ki67 levels are indicators of poor prognosis. Additionally, postoperative adjuvant radiotherapy does not significantly enhance disease-specific survival rates.
		                        		
		                        		
		                        		
		                        	
6.Epidemic characteristics and prediction model analysis of chickenpox in Urumqi in 2014-2019
Zhimeng WANG ; Weiyi FANG ; Yaoqin LU ; Tudi ZULIPIKAER ; Wei CHEN ; Yilihamu SENAWAER ; Kailun ZHANG
Journal of Public Health and Preventive Medicine 2025;36(1):53-56
		                        		
		                        			
		                        			Objective  To construct an optimal prediction model of chickenpox in Urumqi, and to provide reference for formulating the prevention and control strategies of chickenpox.  Methods The multivariate autoregressive moving average model (ARIMAX) and random forest model (RF) were established based on the monthly incidence of chickenpox in Urumqi from 2014 to 2018, and the monthly incidence of chickenpox in 2019 was used to test the models and evaluate their prediction effect. The prediction performance of the two models was compared, and the best model was selected to predict the incidence of chickenpox in Urumqi.  Results  The incidence of chickenpox in Urumqi showed a regular bimodal distribution with obvious seasonality, and it showed a slow upward trend from July 2014 to December 2019. The fitting model was ARIMA(0,1,0)(0,1,1)12, the root mean square error (RMSE) and mean absolute error (MAE) of ARIMAX model training set were 1.29 and 0.95, respectively, and the RMSE and MAE of the test set were 1.88 and 1.44, respectively. The training set RMSE and MAE of RF model were 1.56 and 1.56, respectively, and the test set RMSE and MAE were 4.83 and 3.96, respectively.  Conclusion  The performance of ARIMAX model is better than that of RF model, which can better predict the incidence trend of chickenpox in Urumqi. It is necessary to optimize the prediction model according to the actual situation and provide scientific guidance for the prevention and control of chickenpox.
		                        		
		                        		
		                        		
		                        	
7.Herbal Textual Research on Picrorhizae Rhizoma in Famous Classical Formulas
Feng ZHOU ; Yihan WANG ; Yanmeng LIU ; Xiaoqin ZHAO ; Kaizhi WU ; Cheng FENG ; Wenyue LI ; Wei ZHANG ; Wentao FANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):228-239
		                        		
		                        			
		                        			This article systematically analyzes the historical evolution of the name, origin, quality evaluation, harvesting, processing and other aspects of Picrorhizae Rhizoma by referring to the medical books, prescription books, and other documents of the past dynasties, combined with relevant modern research materials, in order to provide a basis for the development and utilization of famous classical formulas containing this medicinal herb. The research results indicate that Picrorhizae Rhizoma was first recorded in New Revised Materia Medica from the Tang dynasty. Throughout history, Huhuanglian has been used as its official name, and there are also aliases such as Gehu Luze, Jiahuanglian and Hulian. The main source of past dynasties is the the rhizomes of Picrorhiza kurrooa and P. scrophulariiflora. In ancient times, Picrorhizae Rhizoma was mainly imported by foreign traders via Guangzhou and other regions, and also produced in China, mainly in Xizang. In ancient times, it was harvested and dried in early August of the lunar calendar, while in modern times, it is mostly harvested from July to September, with the best quality being those with thick and crispy rhizomes without impurities, and bitter taste. Throughout history, Picrorhizae Rhizoma was collected, washed, sliced, and dried before being used as a raw material for medicine, it has a bitter and cold taste, mainly used to treat bone steaming, hot flashes, infantile chancre fever, and dysentery. There is no significant difference in taste and efficacy between ancient and modern times. Based on the research results, it is recommended that the rhizomes of P. scrophulariiflora in the 2020 edition of Chinese Pharmacopoeia, or the rhizomes of P. kurrooa, can be used in famous classical formulas containing this medicinal herb, which can be processed according to the processing requirements marked by the original formula. For those without clear processing requirements, the dried raw products are used as medicine. 
		                        		
		                        		
		                        		
		                        	
8.Explore of nanopore sequencing technology in ambiguities of HLA genotyping
Nanying CHEN ; Wei ZHANG ; Lina DONG ; Fang WANG ; Yizhen HE ; Chen CHEN ; Faming ZHU
Chinese Journal of Blood Transfusion 2025;38(3):309-315
		                        		
		                        			
		                        			[Objective] To resolve the ambiguities of HLA genotyping generated by next generation sequencing (NGS) using nanopore sequencing technology. [Methods] A total of 38 samples with ambiguous HLA genotyping by NGS in our laboratory were collected, and HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, -DPA1 and -DPB1 loci in these samples were amplified using primers in the same commercial NGS HLA genotyping kit, then subjected to third-generation library construction, and sequenced on the nanopore sequencer. The sequencing data were converted into Fastq files and analyzed by software, and the genotypes of 11 HLA loci were obtained. The ambiguities were counted directly. [Results] The high-resolution genotyping at the second domain of 11 HLA loci of 38 samples using the third generation sequencing (TGS) were consistent with the results of the NGS method at a rate of 100%. The genotypes for the HLA-A, -B, -C, -DRB3, -DRB4, -DQA1 and -DPA1 loci by TGS were all only one result, and the discrimination rate for ambiguities of the HLA-A, -B, -C, and -DQA1 loci (all caused by the difficulty in phasing due to the short NGS read length) was 100%. Among the HLA-DRB1, -DRB5, -DQB1 and -DPB1 loci, the discrimination rate of TGS for the ambiguities caused by non-amplification of exon 1 was 0% and by the short NGS read length was 100%. [Conclusion] Nanopore technology was used to identify the ambiguities of 11 HLA loci in this study, and the ambiguities caused by the short read length disadvantage of the NGS method could be solved effectively and the accuracy of HLA genotyping would be improved.
		                        		
		                        		
		                        		
		                        	
9.Clinical application value of intracavitary PRP infusion combined with IVF-FET in patients with chronic endometritis
Xiaotong ZHANG ; Xiaoyuan HAO ; Rui FANG ; Shuyao HU ; Linkun MA ; Yaqi ZHAO ; Wei HAN
Chinese Journal of Blood Transfusion 2025;38(3):382-387
		                        		
		                        			
		                        			 [Objective] To evaluate the clinical application value of intrauterine perfusion with platelet-rich plasma (PRP) combined with in vitro fertilization-frozen-thawed embryo transfer (IVF-FET) in patients with chronic endometritis (CE). [Methods] A randomized controlled trial (RCT) was conducted, enrolling 60 CE patients undergoing artificial cycle frozen embryo transfer at our hospital from January 2022 to January 2024. Participants were randomly divided into three groups: Group A (routine frozen embryo transfer, n=20), Group B (routine frozen embryo transfer + one PRP intrauterine perfusion, n=20), and Group C (routine frozen embryo transfer + two PRP intrauterine perfusions, n=20). Endometrial thickness during the transformation and transplantation phases, uterine artery pulsatility index (PI), resistance index (RI), systolic peak velocity/end-diastolic velocity (S/D) ratio during transplantation, serum levels of IL-2, IL-4, IL-6, IL-10, and TNF-α during transplantation, as well as biochemical pregnancy rate, clinical pregnancy rate, live birth rate, and early miscarriage rate were compared across groups. [Results] No significant differences in endometrial thickness were observed among the three groups during the transformation phase (P>0.05). During the transplantation phase, endometrial thickness in Groups C and B was significantly higher than in Group A[9.54 (8.96-10.22) and 8.90 (8.34-9.72) vs 8.37 (7.89-8.75) mm, P<0.05], with Group C showing greater thickness than Group B (Z=3.733, P<0.05). Endometrial thickness in Groups C and B during transplantation was significantly increased compared to their respective transformation phases (Z=2.191, 2.462; P<0.05). Groups C and B exhibited lower PI, RI, and S/D values than Group A[PI:1.87 (1.77-1.97), 1.94 (1.88-2.15) vs 2.43 (2.35-2.49); RI:0.75 (0.73-0.77), 0.78 (0.75-0.81) vs 0.84 (0.83-0.86); S/D:2.61 (2.33-3.42), 3.01 (2.20-3.93) vs 3.72 (3.06-4.49); P<0.05]. Group C demonstrated lower PI and RI than Group B (P<0.05). IL-2 levels in Groups C and B were higher than in Group A[3.88 (2.71-5.01), 3.59 (2.73-4.38) vs 3.16 (2.11-3.25) ng/L, P<0.05], while IL-4, IL-6, IL-10, and TNF-α levels were significantly lower (IL-4: Z=1.428, 2.421; IL-6: Z=1.754, 2.435; IL-10: Z=1.754, 2.854; TNF-α: Z=1.961, 1.765; P<0.05). Group C had lower IL-6 levels than Group B (Z=3.976, P<0.05). Biochemical pregnancy rate, clinical pregnancy rate, and live birth rate in Group C were significantly higher than in Group A (75% vs 40%, 70% vs 35%, 60% vs 20%, P<0.05). No significant differences in early miscarriage rates were observed among the groups (χ2=3.750, P>0.05). [Conclusion] Intrauterine autologous PRP perfusion in CE patients enhances pregnancy and live birth rates, improves pregnancy outcomes post-FET, and demonstrates superior efficacy in endometrial repair and receptivity with two PRP perfusions compared to a single perfusion. This provides a safe and effective therapeutic option for optimizing outcomes in CE patients with prior implantation failure.
		                        		
		                        		
		                        		
		                        	
10.Quality evaluation of Mongolian medicine Sendeng-4 based on qualitative and quantitative analysis combined with chemical pattern recognition
Fengye ZHOU ; Jun LI ; Qian ZHANG ; Rongjie LI ; Wei ZHANG ; Jing LIU ; Fang WANG ; Shengnan LI
China Pharmacy 2025;36(9):1040-1045
		                        		
		                        			
		                        			OBJECTIVE To evaluate the quality of Mongolian medicine Sendeng-4 based on qualitative and quantitative analysis combined with chemical pattern recognition, in order to provide the reference for its quality control. METHODS The chemical components in Sendeng-4 were analyzed qualitatively by HPLC-Q-Exactive-MS. The contents of 16 components (methyl gallate, ethyl gallate, epicatechin, dihydromyricetin, genipin-1-O-β-D-gentiobioside, caffeic acid, catechin, corilagin, deacetylasperulosidic acid methyl ester, rutin, geniposide, luteolin, myricetin, quercetin, ferulic acid, and toosendanin) in 15 batches of Sendeng-4 (sample S1-S15) were quantitatively analyzed by HPLC-MS/MS. Cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis were conducted and variable importance projection (VIP) value greater than 1 was used as the index to screen the differential components. RESULTS A total of 73 chemical components were identified in Sendeng-4, including 20 flavonoids, 16 tannins, 14 organic acids, etc. According to the quantitative analysis, the results exhibited that the average contentsthe of above 16 components in 15 batches of Sendeng-4 were 3.683-7.730, 2.391-6.952, 2 275.538-4 377.491, 2 699.188-3 537.924, 858.266-1 377.393, 3.366-11.003, 140.624-315.683,414.629-978.334, 285.501-1 510.457, 27.799-48.325, 3 625.415-6 309.563, 0.506-0.656, 442.337-649.283, 47.093-59.736, 12.942-15.822, 127.738-326.649 μg/g, respectively. According to the results of CA and PCA, 15 batches of samples could be clustered into two categories: S1-S3, S5-S6, S9-S10 and S13 were clustered into one category; S4, S7-S8, S11-S12, S14-S15 were clustered into one category. VIP values of geniposide, epicatechin, deacetylasperulosidic acid methyl ester and genipin-1-O- β-D-gentiobioside were all greater than 1. CONCLUSIONS HPLC-Q-Exactive-MS and HPLC-MS/MS techniques are employed for the qualitative and quantitative analysis of Sendeng-4. Through chemical pattern recognition analysis, four differential components are identified: geniposide, epicatechin, deacetylasperulosidic acid methyl ester, and genipin-1-O-β-D-gentiobioside.
		                        		
		                        		
		                        		
		                        	
            

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