1.SRSF2 promotes glioblastoma cell proliferation by inducing alternative splicing of FSP1 and inhibiting ferroptosis
Dan HUA ; Xuexia ZHOU ; Qian WANG ; Cuiyun SUN ; Cuijuan SHI ; Wenjun LUO ; Zhendong JIANG ; Shizhu YU
Chinese Journal of Pathology 2024;53(5):430-438
Objective:To investigate the effect of serine/arginine-rich splicing factor 2 (SRSF2) on ferroptosis and its possible mechanism in glioblastoma cells.Methods:The online database of gene expression profiling interactive analysis 2 (GEPIA 2) and Chinese Glioma Genome Atlas were used to analyze the expression of SRSF2 in glioblastoma tissue and its association with patients prognosis. To validate the findings of the online databases, the pathological sections of glioblastoma and non-tumor brain tissues from Tianjin Medical University General Hospital, Tianjin, China were collected and analyzed by using immunohistochemistry. Silencing SRSF2 gene expression in glioblastoma cells by siRNA was analyzed with Western blot. The proliferation index was detected by using CCK8 assay. The rescued experiment was conducted by using expression plasmid of pcDNA3.1(+)-SRSF2. The activity of ferroptosis was assessed by using the levels of iron ions and malondialdehyde in glioblastoma cells and the changes in the ratio of glutathione to oxidized glutathione. The changes of gene expression and differential pre-mRNA alternative splicing (PMAS) induced by SRSF2 were monitored by using the third-generation sequencing technology analysis, namely Oxford nanopore technologies (ONT) sequencing analysis.Results:SRSF2 expression was higher in glioblastoma tissues than non-tumor brain tissues. Immunohistochemistry also showed a positive rate of 88.48%±4.60% in glioblastoma tissue which was much higher than the 9.97%±4.57% in non-tumor brain tissue. The expression of SRSF2 was inversely correlated with overall and disease-free disease survivals ( P<0.01). The proliferation index of glioblastoma cells was significantly reduced by silencing with SRSF2 siRNA ( P<0.01) and could be reversed with transfection of exogenous SRSF2. The levels of intracellulariron ions and malondialdehyde increased ( P<0.05), but the glutathione/oxidized glutathione ratio and the expression of key proteins in the glutathione pathway remained unchanged ( P>0.05). ONT sequencing results showed that silencing SRSF2 in glioblastoma cells could induce a significant alternative 3' splice site change on ferroptosis suppressor protein 1 (FSP1). Conclusion:SRSF2 inhibits the ferroptosis in glioblastoma cells and promotes their proliferation, which may be achieved by regulating FSP1 PMAS.
2.Finite element analysis of absorbable screws in Bernese periacetabular osteotomy
Hui CHENG ; Xiaoying MA ; Ningtao REN ; Zhendong ZHANG ; Haigang JIA ; Yong LI ; Jianli ZHANG ; Dianzhong LUO ; Hong ZHANG ; Wei CHAI
Chinese Journal of Orthopaedics 2024;44(6):419-426
Objective:To investigate the stability and feasibility of using absorbable screws during Bernese periacetabular osteotomy.Methods:A retrospective analysis was conducted on a 36 year-old woman diagnosed with developmental dysplasia of the hip, who had undergone Bernese periacetabular osteotomy. Finite element analysis was used to simulate the stability of the acetabulum under loads of 10%, 20%, 50%, and 100% of the patient's weight. The structural stiffness of the pelvis and the maximum equivalent stress on the absorbable screws were observed under different conditions, including whether the acetabular bone block and the ilium were in contact, whether 3 or 4 screws were used, and whether a graft (including fibular cortical bone and PEEK grafts) was used.Results:The structural stiffness of the pelvis fixed with four screws increased by 67%-94% compared to that with three screws. After using a graft, the structural stiffness of the pelvis increased by 50%-83%. As the load increased, the maximum equivalent stress on the screws also increased. When the acetabular bone block and the ilium had no contact, no graft was used, and only three screws were used for fixation, the maximum equivalent stress could reach 518.9 MPa, while this value dropped to 61% when four screws were used (318.7 MPa). When the acetabular bone block and the ilium were in contact, the maximum equivalent stress was about 12% of that when there was no contact, regardless of the number of screws used. When a cortical bone graft or a PEEK graft was used, the maximum equivalent stress could drop to 21%-26% of that without a graft. When the screw strength was 130 MPa, a load of 20% of body weight was applied, and only three screws were used without a graft, the equivalent stress could exceed the strength of the screw; if four screws were used, the equivalent stress was slightly higher than the strength of the screw when a load of 50% of body weight was applied. However, when a graft was used (either cortical bone or PEEK), even when a load of 100% of body weight was applied, the equivalent stress was slightly lower than the strength of the screw.Conclusion:Absorbable screws can provide sufficient stability for Bernese periacetabular osteotomy. The contact between the acetabular bone block and the ilium, an increase in the number of screws, and the use of grafts (cortical bone and PEEK grafts) can further improve stability. Therefore, absorbable screws have broad application prospects in Bernese periacetabular osteotomy.
3.Design and application of "1+3" management module for medical high-value consumables in Operation Room
Junhua ZHANG ; Ming XIAO ; Wenzhi CAI ; Wei LUO ; Lingwu CHEN ; Hong WANG ; Zhendong PEI ; Junyan YAO ; Juan XIAO
Chinese Journal of Modern Nursing 2024;30(13):1720-1723
Objective:To establish the "1+3" management module of high-value consumables in Operation Room and verify its application, so as to provide new ideas for cost management of consumables in Operation Room.Methods:The Operating Room team of Shenzhen Hospital of Southern Medical University designed a "1+3" management module in 2022, where "1" referred to the management process of high-value consumables in Operation Room, and "3" referred to the precise management of consumables in Operation Room warehouse, the management of closed-loop use of Operation Room consumables and adverse event management of consumables. Surgeries using high-value consumables in the Thoracic Surgery Department, Gastrointestinal Surgery Department, and Urology Department of the hospital were selected as the research objects. The surgeries using conventional consumables from January to June 2022 were set as the control group, and the surgeries implementing the "1+3" management module from July to December 2022 were set as the observation group. The number of consumables received by the itinerant nurses before the operation and the number of high-value consumables returned after the operation were compared between the two groups. And the number of missed and error charges for high-value consumables in the two groups were counted and compared.Results:The number of consumables received before operation in the control group was higher than that in the observation group, and the difference was statistically significant ( P<0.05). The number of high-value consumables returned in the observation group was less than that in the control group, and the difference was statistically significant ( P<0.01). The proportion of missed charges for consumables in the observation group was lower than that in the control group, and the difference was statistically significant ( P<0.01), but there was no statistically significant difference in the proportion of incorrect charges between the two groups ( P>0.05) . Conclusions:The "1+3" management module for high-value consumables in Operation Room makes the process of receiving, returning, and charging high-value consumables clear, with traceable data, achieving refined management of high-value consumables in Operation Room, reducing the number of high-value consumables returned to the warehouse and reducing the proportion of missed consumables, which is conducive to effective cost control in Operation Room.
4.Performance evaluation of automated magnetic beads extraction method for the measurement of catecholamine metabolites analyzed by liquid chromatography tandem mass spectrometry
Songlin YU ; Xiaoli MA ; Jialei YU ; Ming LI ; Yacong GUO ; Zhendong LIU ; Yutong ZOU ; Danchen WANG ; Wei LUO ; Ling QIU
Chinese Journal of Laboratory Medicine 2022;45(3):268-274
Objective:To evaluate the performance of magnetic beads extraction method (MGE) for the measurement of catecholamine metabolites by liquid chromatography tandem mass spectrometry.Methods:This is a methodological evaluation study. The linearity, limit of quantitation, recovery, precision, and matrix effect of catecholamine metabolites 3-methoxyepinephrine (MN), 3-methoxynorepinephrine (NMN) and 3-methoxytyramine (3-MT) extracted by MGE method were evaluated according to CLSI C62-A. Consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories and other guidelines, 132 clinical residual plasma samples were collected and extracted by automated MGE and traditional solid phase extraction (SPE) method to compare the harmonization of the two extraction methods.Results:The linearity of MN, NMN and 3-MT extracted by automated MGE was>0.99, and the LOQ for MN, NMN and 3-MT were 0.033 5 nmol/L, 0.054 7 nmol/L and 0.011 0 nmol/L, respectively. The repeatability of MN, NMN and 3-MT were 1.3%-5.1%, 2.2%-5.6% and 1.7%-7.1%, respectively. The total imprecision in the laboratory were 1.5%-8.2%, 2.2%-7.7%, 2.1%-11.2%. Although the absolute recovery is low, the average relative recoveries of MN, NMN and 3-MT were 91.5%-108.5%, 92.0%-108.6%, and 89.3%-104.1%, respectively, and the percentage deviation from the expected concentration was within 15%. After isotope internal standard correction, the relative matrix effect is close to 100%, which can compensate for the potential matrix effect. The results of MGE and SPE of MN, NMN and 3-MT showeda good correlation (correlation coefficient r>0.99). The average relative deviations of MN, NMN and 3-MT were 0.2%, -1.4% and 1.0%, respectively. Conclusion:The automatic MGE method hasa good performance in extracting catecholamine metabolites, and is expected to be used in high-throughput analysis of samples in clinical in the future.
5.Correlation Between Functional Components in Codonopsis pilosula Roots and Soil Factors
Tian-zeng NIU ; Yu-long WANG ; Qin-wen HOU ; Shu-xian XIAO ; Hong-mei LUO ; A-ke LIU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(11):164-172
ObjectiveTo explore the correlation between the content of 4 functional components in Codonopsis pilosula roots from different areas and soil factors, and thereby to lay a theoretical basis for soil ecological regulation and improvement of quality of C. pilosula roots. MethodThe content of lobetyolin, atractylenolide Ⅲ, alcohol extract, and polysaccharides, as well as soil fertility and 16 soil factors in 24 batches of samples from different producing areas were determined. Principal component analysis (PCA) and Pearson's correlation analysis were used to explore the key soil factors leading to the variation of chemical component content in C. pilosula roots. ResultThe content of lobetyolin and atractylenolide Ⅲ in samples from Longxi was the highest, and the content of polysaccharides peaked in samples from Huguan. The content of lobetyolin was in positive correlation with soil total nitrogen, total phosphorus, total potassium, alkali-hydrolyzable nitrogen, and available potassium (P<0.01), as well as soil organic matter, pH, available manganese, and available zinc (P<0.05). There was a positive correlation between pH and atractylenolide Ⅲ content (P<0.05). Soil total potassium was in positive correlation with alcohol extract and polysaccharide content (P<0.01). Soil available zinc was positively correlated with alcohol extract and the polysaccharide content (P<0.05). Sample sites with higher PCA scores were Pingshun, Huguan, and Longxi, which were significantly positively correlated with the content of polysaccharides in C. pilosula roots in different habitats. ConclusionThe content of functional components in C. pilosula roots can be improved by raising soil organic matter content and applying specific fertilizers.
6.Anti-inflammatory Effects of Yuyang Capsule on Bacterial Dermatitis Model Rats and Its Effect on TLR/NF-κB Signaling Pathway
Luncai LUO ; Yan TONG ; Xingguo ZHANG ; Bo CHEN ; Zhendong ZHONG
China Pharmacy 2021;32(4):459-466
OBJECTIVE:To study the anti-inflammatory effects of Yuyang capsule on bacterial dermatitis model rats and its effect on TLR/NF-κ B pathway. METHODS:Minimum inhibition concentration (MIC)and minimal bactericidal concentration (MBC)of Yuyang capsule against Staphylococcus aureus were determined by microdilution test. Totally 50 SD rats were randomly divided into model group ,positive control group (amoxicillin and clavulanate potassium ,120 mg/kg as amoxicillin ),Yuyang capsules high-dose ,medium-dose and low-dose groups (according to MIC ),with 10 rats in each group. The model of bacterial dermatitis was established by using the burned skin of rats infected with S. aureus . 24 h after modeling ,administration groups were intragastrically given the corresponding drug ,and model group was intragastrically given the same amount of normal saline ,once a day,for consecutive 7 days. The skin healing rate was calculated on the 1st,3rd,5th and 7th day of administration ,and the scab formation,decrustation and healing were recorded. The contents of IL- 1β,IL-6,IL-10,TNF-α,hydroxyproline(HYP),collagen Ⅰ(Col Ⅰ)and Col Ⅲ in skin tissue were detected by ELISA. Morphology changes of skin tissues were observed by HE staining. The ultrastructure was observed by transmission electron microscope. Protein expression of TLR 2,TLR4 and p-NF-κB p65 were detected by Western blotting assay. RESULTS :MIC and MBC of Yuyang capsule were 25 mg/mL and 50 mg/mL,respectively. Dose of Yuyang capsules high-dose ,medium-dose and low-dose groups were set at 600,300,150 mg/kg. Compared with model group,scab appeared on the injured skin 3 days after administration in Yuyang capsule high-dose and medium-dose groups ,and decrustation appeared on the injured skin of part mice 5-7 days after administration ;the skin healing rate of the positive control group,Yuyang capsule high-dose and medium-dose groups were all significantly increased at each time point. The contents of IL- 1 β,IL-6,IL-10 and TNF-α,pathological score ,protein expression of TLR 2,TLR4 and p-NF-κB p65 were decreased significantly (P<0.05 or P<0.01);the pathological changes such as inflammatory cell infiltration in skin tissue were improved. The contents of HYP,Col Ⅰ and Col Ⅲ were increased significantly in positive control group and Yuyang capsule high-dose group (P<0.05 or P<0.01). There was no statistical significance in most above indexes in positive control group ,Yuyang capsule high-dose and medium-dose groups (P>0.05). CONCLUSIONS :Yuyang capsule can promote skin healing of bacterial dermatitis model rats and shows certain anti-inflammatory effects ;the mechanism may be related to inhibiting TLR/NF-κB signaling pathway and reducing inflammatory response.
7.Safety evaluation of heavy metals contaminated Astragalus membranaceus using health risk assessment model.
Jiao-Jiao YAO ; Dan-Dan KONG ; Jiao-Yang LUO ; Wen-Jie QIN ; Xue-Mei QIN ; Zhuo-Wen FAN ; Mei-Hua YANG
China Journal of Chinese Materia Medica 2019;44(14):3094-3099
To evaluate the safety of heavy metals contaminated Astragalus membranaceus,an appropriate protocol was established to study the heavy metals pollution level by health risk assessment. This study provided a detailed procedure to assess the medicinal herbs in quality control and safety evaluation,and expected to create awareness among the public on the safety of consuming of A. membranaceus or any other kinds of medicinal herbs. The heavy metals content of Cu,As,Cd,Pb and Hg in a total of 45 batches of A. membranaceus were carefully analyzed with a developed inductively coupled plasma mass spectrometry( ICP-MS). Besides,the heavy metal contamination level was further evaluated through 4 main assessment parameters,including maximum residue limit( MRL) set by International Standard Organization,estimated daily intake( EDI) set by IUPAC,target hazard quotients( THQ) and Total THQ set by USEPA and total THQs in raw herbs of A. membranaceus. In addition,the recommended MRLs of 5 main heavy metals aimed to A. membranaceus were calculated based on the regulated consumption quantity. The result showed that,under the ISO international standard of Chinese medicine-Chinese herbal medicine heavy metals,the unqualified rate was 8. 89% for A. membranaceus,which including 4 batches of A. membranaceus exceeded the MRL of As. Here,the standard THQ value of A. membranaceus was firstly proposed as 0. 02 and 0. 011 25 for adults and children,respectively,which were calculated with the recommended consumption quantity of 30 g and 9 g for adults and children. Furthermore,the values of THQ for As and total THQs in adults and children were exceeded the standard THQ in A. membranaceus,and the recommended MRLs of Pb,Cd,Hg and Cu in above medicinal materials that calculated based on health risk assessment model were higher than the regulated MRLs that set by ISO and Chinese Pharmacopeia. The research showed that the contents of heavy metals in A. membranaceus were not in the safe range and the certain non-carcinogenic risks to human body cannot be neglected. Based on above investigation result,it is easily known that the common evaluation method for raw herbs based on the comparison of MRL of heavy metals was not precise enough,and the international model of health risk assessment should be built for each medicinal herb. Above all,this study provided a more realistic research approach for safety evaluation of any other kinds of heavy metals contaminated medicinal herbs,including the establishment of heavy metals standard limit in a specified medicinal herb under recommended consumption quantity,and it is expected to create awareness among the public on the safety of consuming any other medicinal herbs.
Astragalus propinquus
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chemistry
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Drug Contamination
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Drugs, Chinese Herbal
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analysis
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standards
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Humans
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Metals, Heavy
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analysis
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Plants, Medicinal
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chemistry
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Risk Assessment
8.Survey on pesticide use in Crataegi Fructus and analysis of pesticide residues based on LC-ESI-MS/MS.
Ting-Ting SUN ; Jiao-Yang LUO ; Jia-An QIN ; Kun-Lun LI ; Jin TIAN ; Shi-Hai YANG ; Mei-Hua YANG
China Journal of Chinese Materia Medica 2019;44(23):5088-5093
In order to study the pesticide residues of the medicinal Crataegi Fructus,this study aims to establish an analysis method for pesticide residues( mainly containing insecticides and fungicides) suitable for the actual situation of medicinal Crataegi Fructus based on the survey of the pesticides of the Crataegi Fructus base,combined with the blind screening results of the LC-ESI-MS/MS pesticide screening platform established by the research team in the early stage. Then,the pesticide residues in medicinal Crataegi Fructus from Shandong,Hebei,Henan,Shanxi,and Liaoning( main cultivation areas) were analyzed. The samples were pretreated by the modified Qu ECh ERS method,i.e.,extracted with acetonitrile-water( 9 ∶1),purified by PSA,C_(18),GCB,silica gel. The detection of pesticides was performed by LC-MS/MS. The ion source was ESI with positive scanning mode,and the linearity of 11 kinds of pesticides in the range of 5-300 μg·kg~(-1) was acceptable( R~2>0. 996 9). All the recoveries of pesticides were within 70. 02%~(-1)12. 0% in the low,medium and high levels,with RSD≤17%. The results showed that the detection rate of carbendazim,chlorpyrifos and difenoconazole is 79%,82%,56%,respectively. Besides,the prohibition pesticide carbofuran were detected in some of the batches,indicating the security risk. This study provides methodological references and basic data for risk assessment of Crataegi Fructus and government regulation.
Chromatography, Liquid
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Crataegus/chemistry*
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Drug Contamination
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Drugs, Chinese Herbal/analysis*
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Pesticide Residues/analysis*
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Surveys and Questionnaires
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Tandem Mass Spectrometry
9.Relationship between alterations of cervical sagittal parameters and clinical outcomes after cervical anterior discectomy and fusion
Tao LIU ; Shuiqiang QIU ; Yufeng HUANG ; Shanjin WANG ; Zhendong LUO ; Desheng WU
Chinese Journal of Orthopaedics 2018;38(2):79-85
Objective To explore the relationships between the alterations of cervical sagittal parameters and clinical outcomes after anterior cervical discectomy and fusion (ACDF).Methods From January 2010 to December 2015,a total of 227 patients with cervical spondylosis who undelwent ACDF in Dongfang Hospital affiliated Tongji University were analyzed in this study.There were 109 male and 118 female,with an average age of 52.2± 10.7 years (ranged from 34 to 78 years).The average duration of follow-up after revision surgery was (21.1 ±9.0) months (12-60 months).Comparing with Japanese 0rthopaedic Association (JOA) score and its improvement rate (IR),visual analogue scale (VAS) scores and neck disability index (NDI) between preoperation and 1 year follow-up.Cervical alignment was assessed with the following 3 parameters:T1 slope (T1S),Cobb's angle of C2 to C7 and C2-7 sagittal vertical axis (SVA).And compared the changes of sagittal parameters and clinical efficacy.Results At 1 year follow-up,T1S was increased from 25.4°±8.5° to 27.9°±8.2°,Cobb's angle was increased from 12.7°±8.5° to 15.3°±9.5°,SVA was increased from (21.0± 12.3) mm to (24.30± 11.4) mm,and the differences were statistical significant (P< 0.001).JOA score was increased from (8.5±4.0) points to (13.0±2.4) points at 1 year follow-up,however,VAS score was decreased from 2.7± 1.8 points to 0.2±0.4 points,and NDI was decreased from 48.2%±8.2% to 27.1%± 11.1%,and the differences were statistical significant (P < 0.001).Preoperatively,T1S had positive correlations with both Cobb's angle and SVA before operation (r=0.311,0.213;P=0.000,0.001),but Cobb's angle had negative correlation with SVA (r=-0.246,P=0.000).At 1 year follow-up,T1S had positive correlations with Cobb's angle and SVA (r=0.390,0.392;P=0.000,0.000),and Cobb's angle had negative correlation with SVA (r=-0.131,P=0.048).At 1 year after operation,the change value of Cobb's angle had positive correlation with the change of JOA (r=0.294,P=0.000),but negative correlation with the change of NDI (r=-0.141,P=0.034).Conclusion ACDF is effective in the treatment of cervical spondylosis which the cervical sagittal alignment remains relatively stable,and there were significant correlation between the alteration of cervical sagittal parameters and clinical outcomes after ACDF.
10.Rotational osteotomy on the base of femoral neck in treating osteonecrosis of the femoral head
Kai XIAO ; Dianzhong LUO ; Hui CHENG ; Zhendong ZHANG ; Hong ZHANG
Chinese Journal of Orthopaedics 2018;38(7):425-432
Objective To report the short-term outcomes of the rotational osteotomy on the base of femoral neck in treating osteonecrosis of femoral head.Methods Ten patients (10 hips) with osteonecrosis of femoral head underwent the rotational osteotomy on the base of femoral neck between March 2014 and October 2016.There were 9 males and 1 female.The average age was 29.6 years,ranging from 18 to 42 years.There were 4 patients in right side and 6 in left side.The Association Research Circulation Osseous (ARCO) Stage was from ⅡB to ⅢC (2 cases of ⅡB,2 cases of ⅡC,1 case of ⅢA,1 case of ⅢB,4 cases of ⅢC).The following main procedures were involved in the treatment:surgical hip dislocation,the extension of retinacular soft-tissue flap to protect the main blood supply of the femoral head,anterior or posterior rotational (60°-90°) of femoral head after femoral neck osteotomy.We used three compress cannulated screws to fix the femoral neck osteotomy.The postoperative Harris hip scores (HHS),Western Ontario and McMaster College (WOMAC) score,pelvic A-P view and lateral hip view radiographic characteristics were analyzed to assess the short-term outcomes.Results The average follow-up duration was 16.3 months (7-36 months).No progressive femoral head collapse was observed in 6 patients at the last follow-up.The average post-operative intact ratio was 42.90%± 10.07%.The HHS improved from 80.00±14.93 to 86.83±6.11 (4 cases improved,1 case with no change and 1 case decreased slighdy).The WOMAC score decreased from 16.00±14.53 to 9.00±3.85 (1 cases increased,1 case with no change and 4 case decreased).The remaining 4 cases,however,had the progression of the femoral head collapse.The average post-operative intact ratio was 16.57%±6.57%.The HHS decreased from 77.75±14.66 to 60.75±17.54 (1 case improved slightly and 3 cases decreased).The WOMAC score increased from 12.75±5.06 to 22.50±8.81 (4 cases increased).Conclusion The rotational osteotomy on the base of femoral neck not only shifts the necrosis area away from weight-bearing region but also improves the post-operative intact ratio.Patients with ARCO stage ⅡB-ⅢB may achieve remission of symptoms and improvement of hip function by this surgical technique.However,for patients with ⅢC stage,we should conduct preoperative design to make sure whether the surgery is worth doing or not.Some patients may suffer from post-operative progression collapse of the femoral head.

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