1.Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production
Wang-long CHU ; Tong-jing LI ; Yan SHANGGUAN ; Fang-tao HE ; Jian-fu WU ; Xiu-ping ZENG ; Tao GUO ; Qing-fang WANG ; Fen ZHANG ; Zhen-zhong ZHONG ; Xiao LIANG ; Jun-yuan HU ; Mu-yun LIU
Acta Pharmaceutica Sinica 2024;59(3):743-750
Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.
2.Contralateral mandibular outer cortex "sandwich" bone grafting for correcting lower facial asymmetry with normal occlusal relationship
Guoping WU ; Tong LU ; Yuming QU ; Sheng GAO ; Zhiyang XIE ; Kaili YAN ; Chongxu QIAO ; Shunchao YAN ; Shu WANG ; Wensong SHANGGUAN
Chinese Journal of Plastic Surgery 2024;40(10):1049-1058
Objective:To investigate the clinical outcomes of using autologous mandibular outer cortex "sandwich" grafting to augment mandible for correcting lower facial asymmetry with normal occlusal relationships.Methods:A retrospective analysis was conducted on the clinical data of patients with lower facial asymmetry treated at the Department of Plastic Surgery, the Affiliated Friendship Plastic Surgery Hospital of Nanjing Medical University, from January 2016 to December 2019. Preoperative cone-beam CT (CBCT) scans of the maxillofacial region were obtained to acquire three-dimensional data of the maxillofacial skeleton. Computer-aided design was used to determine the osteotomy range of the mandibular outer cortex and design osteotomy guide template. The outer cortex of the larger mandibular side (donor site) was harvested according to the osteotomy guide template and sectioned. The segmented mandibular outer cortex was then contoured to match the arc of the recipient side’s mandibular outer cortex and fixed to the inner side of the recipient mandibular outer cortex, thus increasing the width and thickness of the expanded mandible. Follow-up was conducted at 7th day and 6 months postoperatively, CBCT scan was performed to measure the changes in ramus height (Co-Go), mandibular body length (Go-Me), and mandibular outer cortex thickness, and volume, and the patient satisfaction with facial appearance (1 to 5 points, with higher scores indicating higher satisfaction), were assessed to evaluate surgical outcomes. Statistical analysis was performed using SPSS 12.0 software. Paired t-tests were used to compare patient satisfaction scores preoperatively and six months postoperatively. Repeated measurement ANOVA was used to compare Co-Go and Go-Me measurements preoperatively, 7th day, and 6 months postoperatively. If a statistically significant difference is found, further analysis using post-hoc testing(Tukey’s HSD test) will be conducted to examine the data. Results:Sixteen patients with lower facial asymmetry were included, comprising of 5 males and 11 females, aged 18 to 40 years, with an average age of 25.2 years. Surgical method included contralateral mandibular outer cortex "sandwich" grafting to the expanded mandible in 9 cases and simultaneous genioplasty in 7 cases. Follow-up ranged from 6 months to 5 years, with an average follow-up of 18.6 months. All patients experienced numbness of the lower lip postoperatively, which resolved within six months, and no severe complications occurred. The symmetry of lower facial contour improved significantly and remained stable. Patient satisfaction score for facial appearance increased from (1.63±0.62) points preoperatively to (4.19±0.75) points six months postoperatively( P<0.01). The differences in Co-Go on the donor side and Go-Me on the recipient side across the three time points(preoperatively, 7th day and 6 months postoperatively) were not statistically significant(all P>0.05). However, the differences in Go-Me on the donor side and Co-Go on the recipient side across the three time points were statistically significant(all P<0.05). On the donor side, the mandibular outer cortex thickness decreased by a maximum of 6 mm on 7th day postoperatively and increased by a maximum of 2 mm at 6 months postoperatively compared to 7th day. On the recipient side, mandibular outer cortex thickness increased by a maximum of 6 mm on 7th day postoperatively and decreased by a maximum of 2 mm at six months postoperatively compared to 7th day. The volume of the mandibular outer cortex on the recipient side increased by (4 415.94±1 017.21)mm 3 at 7th day postoperatively compared to preoperatively, and decreased by (202.63±300.85)mm 3 at 6 months postoperatively. Conclusion:For lower facial asymmetrical with normal occlusal relationships and no occlusal plane deviation, contralateral mandibular outer cortex "sandwich" bone grafting can effectively increase the width and volume of the mandible on the grafted side, achieving favorable clinical outcomes.
3.Contralateral mandibular outer cortex "sandwich" bone grafting for correcting lower facial asymmetry with normal occlusal relationship
Guoping WU ; Tong LU ; Yuming QU ; Sheng GAO ; Zhiyang XIE ; Kaili YAN ; Chongxu QIAO ; Shunchao YAN ; Shu WANG ; Wensong SHANGGUAN
Chinese Journal of Plastic Surgery 2024;40(10):1049-1058
Objective:To investigate the clinical outcomes of using autologous mandibular outer cortex "sandwich" grafting to augment mandible for correcting lower facial asymmetry with normal occlusal relationships.Methods:A retrospective analysis was conducted on the clinical data of patients with lower facial asymmetry treated at the Department of Plastic Surgery, the Affiliated Friendship Plastic Surgery Hospital of Nanjing Medical University, from January 2016 to December 2019. Preoperative cone-beam CT (CBCT) scans of the maxillofacial region were obtained to acquire three-dimensional data of the maxillofacial skeleton. Computer-aided design was used to determine the osteotomy range of the mandibular outer cortex and design osteotomy guide template. The outer cortex of the larger mandibular side (donor site) was harvested according to the osteotomy guide template and sectioned. The segmented mandibular outer cortex was then contoured to match the arc of the recipient side’s mandibular outer cortex and fixed to the inner side of the recipient mandibular outer cortex, thus increasing the width and thickness of the expanded mandible. Follow-up was conducted at 7th day and 6 months postoperatively, CBCT scan was performed to measure the changes in ramus height (Co-Go), mandibular body length (Go-Me), and mandibular outer cortex thickness, and volume, and the patient satisfaction with facial appearance (1 to 5 points, with higher scores indicating higher satisfaction), were assessed to evaluate surgical outcomes. Statistical analysis was performed using SPSS 12.0 software. Paired t-tests were used to compare patient satisfaction scores preoperatively and six months postoperatively. Repeated measurement ANOVA was used to compare Co-Go and Go-Me measurements preoperatively, 7th day, and 6 months postoperatively. If a statistically significant difference is found, further analysis using post-hoc testing(Tukey’s HSD test) will be conducted to examine the data. Results:Sixteen patients with lower facial asymmetry were included, comprising of 5 males and 11 females, aged 18 to 40 years, with an average age of 25.2 years. Surgical method included contralateral mandibular outer cortex "sandwich" grafting to the expanded mandible in 9 cases and simultaneous genioplasty in 7 cases. Follow-up ranged from 6 months to 5 years, with an average follow-up of 18.6 months. All patients experienced numbness of the lower lip postoperatively, which resolved within six months, and no severe complications occurred. The symmetry of lower facial contour improved significantly and remained stable. Patient satisfaction score for facial appearance increased from (1.63±0.62) points preoperatively to (4.19±0.75) points six months postoperatively( P<0.01). The differences in Co-Go on the donor side and Go-Me on the recipient side across the three time points(preoperatively, 7th day and 6 months postoperatively) were not statistically significant(all P>0.05). However, the differences in Go-Me on the donor side and Co-Go on the recipient side across the three time points were statistically significant(all P<0.05). On the donor side, the mandibular outer cortex thickness decreased by a maximum of 6 mm on 7th day postoperatively and increased by a maximum of 2 mm at 6 months postoperatively compared to 7th day. On the recipient side, mandibular outer cortex thickness increased by a maximum of 6 mm on 7th day postoperatively and decreased by a maximum of 2 mm at six months postoperatively compared to 7th day. The volume of the mandibular outer cortex on the recipient side increased by (4 415.94±1 017.21)mm 3 at 7th day postoperatively compared to preoperatively, and decreased by (202.63±300.85)mm 3 at 6 months postoperatively. Conclusion:For lower facial asymmetrical with normal occlusal relationships and no occlusal plane deviation, contralateral mandibular outer cortex "sandwich" bone grafting can effectively increase the width and volume of the mandible on the grafted side, achieving favorable clinical outcomes.
4.Construction and application of a multi-drug resistance bacteria information management system/
Bo WU ; Yuying YAN ; Liying SONG ; Xuan YANG ; Xuejun SHANGGUAN ; Rong LIN
Chinese Journal of Nursing 2024;59(16):1942-1947
Objective To construct and apply a multidrug-resistant bacteria information management system,and evaluate its effectiveness and accuracy in the management of patients with multidrug-resistant bacteria infection.Methods A system construction team was established to develop a multidrug-resistant bacteria information management system,which includes 4 modules:early warning,execution,monitoring,and statistical decision-making.Patients with positive detection of multidrug-resistant bacteria admitted to a tertiary A obstetrics and gynecology hospital in Zhejiang Province from January to December 2022,members of the system construction team,and clinical medical staff were selected as the research subjects.The execution efficiency of multidrug-resistant bacteria management,as well as the system's accuracy,usability,and satisfaction were compared before(January to June 2022)and after(July to December 2022)the application of the system.Results After the implementation of the system,the immediate feedback rate of multidrug-resistant bacteria early warning information increased from 62.87%to 89.78%;the rate of issuing isolation medical orders rose from 61.07%to 93.33%;the accuracy of the implementation of isolation measures for patients increased from 66.67%to 98.01%;all differences are statistically significant(P<0.001).The accuracy rate of the system in making decisions is 88.44%;the usability score given by medical staff for the system is 44.04 points,and the satisfaction score of the system construction team members and medical staff to the system is 121.25 points,both at a high level.Conclusion The multidrug-resistant information management system is equipped with features such as immediate alerts,multi-channel notifications,infection control department supervision,and auxiliary decision-making,which can provide medical staff with accurate decision reports.Preliminary application results show that the system has a high level of accuracy and good usability.
5.Effectiveness of rotational filling of an inferolaterally pedicled tongue-shaped parenchymal flap via a double periareolar incision on breast ptosis
Kaili YAN ; Chongxu QIAO ; Shunchao YAN ; Zai SHI ; Jingyi XU ; Yuming QU ; Shu WANG ; Wensong SHANGGUAN ; Guoping WU
Chinese Journal of Medical Aesthetics and Cosmetology 2024;30(6):582-587
Objective:To evaluate the effectiveness of correcting breast ptosis using the rotational filling of an inferolaterally pedicled tongue-shaped parenchymal flap via a double periareolar incision.Methods:A retrospective study was conducted on 34 female patients (68 breasts) with breast ptosis treated at the Department of Plastic Surgery at the Affiliated Friendship Plastic Surgery Hospital of Nanjing Medical University from January 2016 to June 2023. The patients' ages ranged from 23 to 53 years, with an average of (37.4±7.9) years. All patients underwent correction of breast ptosis using an inferolaterally pedicled tongue-shaped parenchymal flap via a double periareolar incision. Pre-operative and post-operative measurements were taken to assess the corrective outcomes, including mammary base width (MBW), areola diameter (AD), nipple-to-nipple distance (NN), suprasternal notch-to-nipple distance (SSN-N), nipple-to-inframammary fold distance (N-IMF), and thoracic circumference at the nipple level (TN). Patients' satisfaction was assessed using the BREAST-questionnaire (BREAST-Q) module.Results:Postoperative first-stage wound healing was achieved in all 34 patients without complications such as hematoma, infection, or nipple necrosis. The follow-up period ranged from 6 to 60 months, with a median of 12 months. Early postoperative observations showed periareolar folds, which disappeared within 3 to 6 months in 31 cases, while mild periareolar folds, persisted for up to 12 months in 3 cases. Four patients reported reduced nipple sensation postoperatively. Measurements showed a statistically significant increase in TN and a reduction in NN, SSN-N, AD, N-IMF, and MBW compared to preoperative values (all P<0.001). BREAST-Q scores indicated significant improvements in satisfaction with breasts, psychosocial well-being and physical well-being, with preoperative scores of (21.6±8.9), (42.4±11.9) and (49.7±12.9) scores increasing to (82.3±8.5), (81.9±8.1) and (81.2±8.3) scores, respectively (all P<0.001). Conclusions:The double periareolar incision technique with rotational filling of an inferolaterally pedicled tongue-shaped glandular flap is a safe and effective method for correcting breast ptosis.
6.Efficacy of W-genioplasty on the broad and short chin
Zhiyang XIE ; Sheng GAO ; Kaili YAN ; Tong LU ; Chao HU ; Shu WANG ; Wensong SHANGGUAN ; Guoping WU
Chinese Journal of Medical Aesthetics and Cosmetology 2023;29(1):19-22
Objective:To investigate the clinical effect of W-shaped genioplasty in the correction of broad and short chin deformity.Methods:Thirty-eight patients (5 males, 33 females, aged 20 to 41 years, mean 27.3 years) complained with broad and short chin were admitted to the Affiliated Friendship Plastic Surgery Hospitalof Nanjing Medical University from January 2019 to December 2021. CBCT scan and three-dimensional reconstruction were performed to design osteotomy line and determine the distance of chin lengthening, narrowing and advancing or retrocession preoperatively. Under general anesthesia, the W-shaped osteotomy was performed using an intraoral incision, and the angle between the bilateral free bone fragments, the distance of downward and forward movement were adjusted to change the curvature, width, length and prominence of the lower edge of the chin according to the preoperative designs. The results were evaluated by clinical appearances and image analyses at a follow-up of 3-24 months.Results:The amount of vertical lengthening of the chin in 38 cases were 2 mm to 5 mm, with an average of 3.02 mm. The horizontal narrowing width distances were 3-7 mm, with an average of 5.6 mm. The patients were followed up for 3-24 months, with an average of 10.6 months. There were no complications such as hematoma, wound dehiscence, accidental fracture, surgical area infection and permanent neurosensory disorder. 38 patients had transient sensory loss in the lower lip region of varying degrees, but all recovered spontaneously during routine follow-up period. All patients were satisfied with the improvement of facial contour.Conclusions:W-shaped geinoplasty preserves the central bone of the chin and the attachment of genioglossus muscle, which does not affect the normal anatomy and physiological function of the oral cavity. After osteotomy, the bone is removed and the bilateral bone fragments move flexibly. It can effectively change the radian, width, length and protrusion of the lower edge of the chin in three dimensions, so as to correct the wide and short deformity of the chin.
7.Surveillance results of iodine deficiency disorders in key populations in Ganzhou City, Jiangxi Province from 2018 to 2022
Yuekang YAN ; Hongmin CAO ; Jun SHANGGUAN ; Yaoqin ZHU ; Zijuan YANG ; Rongrong YANG ; Gang DU ; Qi HUANG
Chinese Journal of Endemiology 2023;42(11):908-912
Objective:To investigate the status of iodine deficiency disorders in children and pregnant women in Ganzhou City, Jiangxi Province, and to provide a basis for scientific iodine supplementation and adjustment of prevention and control strategies.Methods:According to the requirements of the National Iodine Deficiency Disorders Surveillance Program (2016 Edition), iodine deficiency disorders surveillance was carried out in 18 counties (cities, districts) under the jurisdiction of Ganzhou City from 2018 to 2022. Edible salt and urine samples taken from children aged 8 to 10 and pregnant women were collected to detect salt and urine iodine levels. At the same time, B-ultrasound method was adopted to measure children's thyroid volume, and the goiter rate was calculated.Results:From 2018 to 2022, a total of 27 075 edible salt samples were collected from key populations (children aged 8 - 10 and pregnant women), with a median salt iodine of 25.00 mg/kg and a qualified iodized salt consumption rate of 96.24% (26 057/27 075). The difference in the qualified iodized salt consumption rate among key populations in different years was statistically significant (χ 2 = 29.09, P < 0.001). A total of 18 061 urine samples were collected from children, with a median urine iodine of 192.10 μg/L, there was a statistically significant difference in the urine iodine level of children between different years ( H = 82.59, P < 0.001). A total of 9 014 urine samples were collected from pregnant women, with a median urine iodine of 177.20 μg/L, there was a statistically significant difference in urine iodine level of pregnant women between different years ( H = 78.78, P < 0.001). A total of 8 621 children's thyroid glands were examined, including 34 cases of goiter, with a goiter rate of 0.39%, and the goiter rate showed a decreasing trend year by year (χ 2trend = 11.09, P < 0.001). Conclusions:From 2018 to 2022, the iodine nutrition of children and pregnant women in Ganzhou City is at an appropriate level, the consumption rate of qualified iodized salt (> 90%) and the children's goiter rate (< 5%) met the national iodine deficiency disorders elimination standards. Ganzhou City continues to maintain the status of eliminating iodine deficiency disorders.
8. Sequence analysis and SSR marker mining of a peroxidase gene in Senecio scandens
Chinese Traditional and Herbal Drugs 2019;50(8):1952-1959
Objective: To study the relationship between the sequence structure and function of a peroxidase gene (POD) and mine and confirm SSR locus of this gene in Senecio scandens. Methods: A POD gene was obtained from full-length cDNA library of S. scandens. Then a series of online bioinformatics tools were used to analyze nucleic acid sequence and amino acid structure of this gene. According to its nucleotide sequence, one SSR locus was searched and a pair of primers were designed. The PAGE electrophoresis confirmed that this primer pair had polymorphism among several landraces. Results: The target gene had the structural characteristics of typical PODs. It showed a high conservation in functional domains. PAGE electrophoresis showed that the primer could be stably amplified in different landraces with obvious polymorphisms. Conclusion: The results expanded the understanding of the function and sequence of a POD in S. scandens. The newly developed marker could be effectively applied to the detection of POD gene and breeding projects, and it also provided reference for POD researches of other species.
9.Antepenultimate residue at the C-terminus of NADPH oxidase RBOHD is critical for its function in the production of reactive oxygen species in Arabidopsis.
Qiu-Ying LI ; Ping LI ; Nang MYINT PHYU SIN HTWE ; Ke-Ke SHANGGUAN ; Yan LIANG
Journal of Zhejiang University. Science. B 2019;20(9):713-727
Production of reactive oxygen species (ROS) is a conserved immune response primarily mediated by NADPH oxidases (NOXs), also known in plants as respiratory burst oxidase homologs (RBOHs). Most microbe-associated molecular patterns (MAMPs) trigger a very fast and transient ROS burst in plants. However, recently, we found that lipopolysaccharides (LPS), a typical bacterial MAMP, triggered a biphasic ROS burst. In this study, we isolated mutants defective in LPS-triggered biphasic ROS burst (delt) in Arabidopsis, and cloned the DELT1 gene that was shown to encode RBOHD. In the delt1-2 allele, the antepenultimate residue, glutamic acid (E919), at the C-terminus of RBOHD was mutated to lysine (K). E919 is a highly conserved residue in NADPH oxidases, and a mutation of the corresponding residue E568 in human NOX2 has been reported to be one of the causes of chronic granulomatous disease. Consistently, we found that residue E919 was indispensable for RBOHD function in the MAMP-induced ROS burst and stomatal closure. It has been suggested that the mutation of this residue in other NADPH oxidases impairs the protein's stability and complex assembly. However, we found that the E919K mutation did not affect RBOHD protein abundance or the ability of protein association, suggesting that the residue E919 in RBOHD might have a regulatory mechanism different from that of other NOXs. Taken together, our results confirm that the antepenultimate residue E is critical for NADPH oxidases and provide a new insight into the regulatory mechanisms of RBOHD.
Agrobacterium tumefaciens/metabolism*
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Alleles
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Arabidopsis/metabolism*
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Arabidopsis Proteins/genetics*
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Gene Expression Regulation, Plant
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Genetic Techniques
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Humans
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Lipopolysaccharides/metabolism*
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Luminescence
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Mutation
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NADPH Oxidase 2/chemistry*
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NADPH Oxidases/genetics*
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Plant Stomata/metabolism*
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Protein Domains
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Reactive Oxygen Species/metabolism*
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Nicotiana/metabolism*
10. SSR marker development and genetic diversity analysis of Stephania kwangsiensis
Chinese Traditional and Herbal Drugs 2018;49(24):5910-5915
Objective To develop simple sequence repeat (SSR) molecular markers based on Stephania kwangsiensis transcriptome and to evaluate genetic diversity of S. kwangsiensis in natural populations. Methods The unigenes from S. kwangsiensis transcriptome were used to explore SSR loci. SSR primers were designed by oligo 7.0. Polymorphic primers were selected by electrophoresis and were applied to the investigation of the genetic diversity of S. kwangsiensis. Results A total of 6 833 SSR loci (33.11%) were obtained from 20 637 unigenes in the S. kwangsiensis transcriptome. Of the 50 SSR markers tested, ten pairs of highly polymorphic primers were selected to analyze the genetic polymorphisms of 63 individuals from five populations. Ten primers produced 83 band loci, all (100%) of which were polymorphic, and the polymorphism information content (PIC) was 0.688 9. At the species level and population level, the Nei’ s gene diversity (H) was 0.725 2 and 0.613 4, respectively; the Shannon’s Information index (I) was 1.576 6 and 1.220 3, respectively; The observed heterozygosity (Ho) was 0.584 5 and 0.558 4, respectively; The expected heterozygosity (He) was 0.731 2 and 0.643 5, respectively. The genetic differentiation coefficient (Fst) was 0.146 5 and the gene flow (Nm) was 1.456 9. Based on UPGMA cluster analysis, the five populations of S. kwangsiensis were divided into three clusters. Conclusion The developed SSR markers were applicable to the genetic diversity evaluation of S. kwangsiensis. The genetic diversity of this plant was relatively high at both species level and population level. The genetic differentiation mainly existed within populations.

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