[Clinicopathological and molecular genetic features of neuromuscular choristoma-associated desmoid type fibromatosis].

Objective: To investigate the clinicopathological and genetic characteristics of neuromuscular choristoma-associated desmoid type fibromatosis (NMC-DF). Methods: The clinical morphological and immunohistochemical features of 7 NMC-DF cases diagnosed from January 2013 to January 2023 in Beijing Jishuitan Hospital were retrospectively analyzed. A series of neuromuscular choristoma and neuromuscular choristoma-associated desmoid type fibromatosis were evaluated for CTNNB1 mutations, and hotspot mutations for CTNNB1 were tested in 4 NMC-DF cases using Sanger sequencing. Results: The tumors were collected from 3 females and 4 males, aged 1 to 22 years (mean 7.1 years), involving the sciatic nerve (n=4), brachial plexus (n=2) or multiple nerves (n=1). The course of the disease spanned from 3 months to 10 years. Two cases were recurrent tumors. All the 7 NMC cases showed endoneurial intercalation of mature skeletal muscle fibers among the peripheral nerve fascicles, and the histologic features of the NMC-DF were strikingly similar to the conventional desmoid-type fibromatosis. By immunohistochemistry, all NMC and NMC-DF cases showed aberrant nuclear staining of ß-catenin (7/7), the muscle cells in NMC were intensely immunoreactive for desmin, and the admixed nerve fibers were highlighted by NF and S-100 (7/7). Four NMC and NMC-DF had CTNNB1 mutations, 3 c.121A>G (p.T41A) and 1 c.134C>T (p.S45F). Follow-up of the 7 cases, ranging from 22 to 78 months, showed tumor recurrence in 2 patients at 3 and 8 months respectively after the first surgical resection, of which 1 patient underwent above-knee amputation. No recurrence occurred in other cases with tumor excision and neurological reconstruction surgery. There was no metastasis occurred in the 7 cases. Conclusions: NMC is a rare congenital lesion with differentiated mature skeletal muscle tissue found in peripheral nerve fascicles, and approximately 80% of patients with NMC develop a soft tissue fibromatosis. CTNNB1 mutation in the Wnt signaling pathway may be involved in the pathogenesis of NMC and NMC-DF, and S45F mutations seems to have a higher risk of disease progression.

[Pediatric myofibroma/myofibromatosis of the soft tissue and bone: a clinicopathological analysis of 28 cases].

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of pediatric myofibroma/myofibromatosis of the soft tissue and bone. Methods: All cases of pediatric myofibroma/myofibromatosis of the soft tissue and bone diagnosed between January 2011 and December 2018 were retrieved from the surgical pathology records in the Department of Pathology, Beijing Jishuitan Hospital, Beijing, China. Clinical and radiological data were collected. H&E and immunohistochemistry were used to examine histological and immunophenotypic features and to make the diagnosis and differential diagnosis. The relevant literature was also reviewed. Results: Twenty-eight cases of pediatric myofibroma/myofibromatosis of the soft tissue and bone were respectively collected. The patients' ages ranged from 2 months to 14 years, with a mean age of 7 years. There were 7 females and 21 males. There were 12 cases located in soft tissue, including the finger (n=9), upper arm (n=1) and foot (n=2). There were 14 cases located in the bone of limb, including the femur (n=8), tibia (n=4), clavicle (n=2), fibula (n=2) and radius (n=1). There were 2 cases of myofibromatosis involving multiple bones. Radiology showed lytic lesions in the bone. The proliferation of spindle-shaped myofibroblasts arranged in fascicles with indistinct eosinophilic cytoplasm and bland nuclei, with no pleomorphism and cytological atypia. The characteristic histologic structure was the biphasic nodular growth pattern with cellular and paucicellular regions. The tumors might arrange in a hemangiopericytoma-like pattern. The stroma varied between dense fibrosis and myxoid changes. The reactive new bone formation and inflammatory cell infiltration also existed. Immunohistochemical study showed that the SMA was positive. The surgical resections were performed. One of the patients had tumor recurrence as a result of 11-month follow-up. Conclusions: The pediatric myofibroma/myofibromatosis of the soft tissue and bone is a very rare benign tumor and has a good prognosis. It has a characteristic morphology and its differential diagnosis from other spindle cell tumors could be made with the immunohistochemical analysis.

[Radiation-associated sarcomas of bone and soft tissue: a clinicopathological analysis of 46 cases].

Objective: To investigate the clinical, imaging, histological, and molecular features and the differential diagnosis of radiation-associated sarcomas of bone and soft tissue. Methods: Forty-six cases of radiation-associated sarcomas of the bone and soft tissue in Beijing Jishuitan Hospital from January 2010 to January 2022 were retrospectively analyzed; and the imaging, histological features and immunophenotype were examined. Results: There were 33 females and 13 males, aged from 18 to 74 years, with a mean of 52 years. The most common site of radiation-associated sarcomas were the limbs and spine (15 cases), followed by the chest (9 cases). The primary diseases included epithelial tumors (15 breast cancer, 6 cervical cancer, and 5 bowel cancer), hematolymphoid tumors, bone and soft tissue tumors and infectious lesions. The latent period of radiation-associated sarcomas ranged from 2-22 years, with an average of 11.6 years. Histopathologically, the morphology was divergent from the primary tumor. The most common malignant tumor type was undifferentiated sarcoma (22 cases), followed by osteosarcoma (16 cases). The immunophenotype of radiation-related sarcoma was almost the same as the corresponding soft tissue sarcoma. Conclusions: Radiation-induced sarcoma has a wide range of primary tumor types and its imaging, morphology and immunohistochemical features are similar to those of the primary sarcoma of bone and soft tissue. Clinical correlation is often recommended for the differential diagnosis.

[Development of an asthenopia survey questionnaire for general surveys].

Objective: To design a visual fatigue questionnaire that can be used for population surveys. Methods: This was a cross-sectional study that involved three stages of subjects' recruitment. In the first stage, by convenience sampling, 150 individuals who complained of visual fatigue were selected at public places in Wenzhou City in May 2016. The 19-Item Asthenopia Survey Questionnaire (ASQ-19) was used to conduct the survey, and the questionnaire was adjusted. In the second stage, 200 outpatient participants were recruited from Wenzhou Medical University Affiliated Eye and Optometry Hospital from June 2016 to May 2017 and were divided into a visual fatigue group and a control group based on clinical diagnosis. The adjusted visual fatigue questionnaire was used for validation. In the third stage, 64 outpatient participants who met the inclusion criteria were continuously recruited from the Wenzhou Medical University Affiliated Eye and Optometry Hospital in July 2022. They were tested using the adjusted visual fatigue questionnaire and retested one week later. During the questionnaire adjustment stage, factor analysis and feedback were used to adjust the scoring method and items of the ASQ-19 questionnaire. The adjusted questionnaire was then analyzed for reliability, validity, accuracy, and subject acceptance during the validation and retest stages. Results: A total of 403 participants were included, and 456 questionnaires were distributed. Eventually, 432 valid questionnaires were collected from 379 participants, resulting in a valid response rate of 94.7%. During the questionnaire adjustment phase, there were 140 valid questionnaires from 140 participants consisting of 56 males and 84 females with an average age of (35.2±12.4) years. In the questionnaire validation phase, there were 186 valid questionnaires from 186 participants. Sixty-two participants had visual fatigue and 124 were controls. During the questionnaire retesting phase, 53 participants yielded 106 valid questionnaires. The group consisted of 20 males and 33 females with an average age of (22.8±4.9) years. After factor analysis, the symptom severity graded as none, mild, moderate, severe, and very severe was scored as 0, 1, 2, 3, and 4 points, respectively. The total score was 44, and the final questionnaire consisted of 11 items (numbered 1, 2, 3, 5, 6, 8, 10, 15, 17, 18, and 19). The 11-Item Asthenopia Survey Questionnaire (ASQ-11) had a Cronbach's α coefficient of 0.89, a split-half reliability of 0.82, and a test-retest Pearson correlation coefficient of 0.90 (P<0.001). The structural validity was 51.26%, and the discriminative validity was a t-value of 9.19 (P<0.001). On average, it took (2.82±0.43) minutes for participants to complete the questionnaire. The receiver operating characteristic curve had a cutoff value of 8.5, with a sensitivity of 74.19% and a specificity of 80.65%. Conclusion: The ASQ-11, with fewer items and a shorter completion time, is easy for participants to use and is suitable for screening or self-assessment of visual fatigue in the general population. Additionally, it is convenient for clinical and epidemiological studies related to visual fatigue.

[The diagnostic value of CCNB3 and BCOR expression in BCOR-CCNB3 sarcoma].

Objective: To investigate the diagnostic value of expression of CCNB3 and BCOR in BCOR-CCNB3 sarcoma (BCS). Methods: Fifteen cases of BCS confirmed by fluorescence in situ hybridization (FISH) and/or reverse transcription-polymerase chain reaction (RT-PCR) from January 2014 to October 2021 at Beijing Jishuitan Hospital were collected. Immunohistochemical EnVision method was used to detect the expression of CCNB3 and BCOR in 15 cases of BCS and in 65 non-BCS tumors (54 cases of Ewing's sarcoma, 5 cases of CIC rearranged sarcoma, 4 cases of synovial sarcoma, 1 case of mesenchymal chondrosarcoma and 1 case of soft tissue clear cell sarcoma). Results: Immunohistochemical staining for CCNB3 revealed strongly diffuse nuclear staining in 14 of 15 (14/15) BCS cases, whereas none of the 65 non-BCS tumors showed any staining. Immunohistochemical staining for BCOR showed strongly diffuse nuclear staining in 11 (11/14) BCS cases; seven of the 65 (7/65, 10.8%) non-BCS tumors showed variable staining (five cases of Ewing sarcoma, one cases of synovial sarcoma, and one case of mesenchymal chondrosarcoma). The sensitivity and specificity of CCNB3 in diagnosing BCS were 93.3% and 100% and these of BCOR were 78.6% and 89.2%, respectively. Conclusions: CCNB3 is a highly sensitive and specific marker for BCS.The antibody may help screening BCS.

Comparing responses of dairy cows to short-term and long-term heat stress in climate-controlled chambers.

Heat stress (HS) in dairy cows can be classified into short-term heat stress (STHS) and long-term heat stress (LTHS) according to the number of consecutive days in HS. The comparative study of these 2 types of HS is limited in terms of their effects on the production and energy metabolism of cows. In this study, 4 lactating Holstein cows (102.5 ± 12 days in milk, 605 ± 22 kg of body weight, second parity) fitted with rumen fistulae were randomly assigned to 1 of 2 groups in a 2 × 2 crossover design and allocated to 1 of 2 climate-controlled chambers. This study contained 2 periods, each with a control phase and a HS phase. There was a recovery phase between 2 periods. The HS phase comprised either STHS (3 d) or LTHS (7 d) treatments. Data collected from the 3 d of STHS and the last 3 d of LTHS were compared. The chambers were set at thermal neutral conditions (20°C, 50% humidity) during the control and recovery phases or cyclical HS conditions (26-38°C, 50% humidity) during the HS phase. Compared with STHS, LTHS decreased milk yield by 17.2% and dry matter intake by 12.6%, indicating that LTHS caused a more severe decline in milk production and feed intake. In addition, LTHS decreased milk protein concentration by 6.8% and milk protein yield by 22.4%. In comparison with STHS, LTHS decreased rumen liquor volatile fatty acid (29.7%), blood glucose (11.6%), and nonesterified fatty acid (13.6%) concentrations, but increased milk urea nitrogen by 15.1%, blood urea nitrogen by 8.6%, and creatine concentrations by 15.4%. Our results suggest that although reduced feed intake may be mainly responsible for reduced milk production during STHS, impaired rumen metabolism and suppressed mobilization of adipose tissue could be the main reasons for further reduction in milk yield during LTHS.

[DNA sequencing of H3F3A mutations in H3.3 immunohistochemistry-negative giant cell tumors of bone].

Objective: To investigate the subtypes of H3F3A DNA mutation in H3.3 immunohistochemistry (IHC) negative giant cell tumors of bone (GCTB). Methods: IHC expression of G34W mutated protein was evaluated in 181 cases GCTB. In H3.3 IHC negative cases, Sanger DNA sequencing analysis was used to detect the subtypes H3F3A mutations. Results: Overall, 164 (90.61%) cases of GCTB showed nuclear expression of H3.3, and 17 cases were negative. These 17 H3.3 negative cases were subjected to Sanger DNA sequencing analysis; results showed that eight presented rare mutation subtypes occurring at glycine 34 to leucine (G34L, 3/181, 1.66%), glycine 34 to valine (G34V, 3/181, 1.66%) and glycine 34 to arginine (G34R, 2/181, 1.10%), and the other nine cases were wild type (glycine 34, 9/181, 4.97%). Sanger DNA sequencing analysis confirmed the absence of G34W mutation in the H3.3 negative cases. Combining IHC and DNA sequencing analysis increased the detection rate of H3F3A mutation in the GCTB to 95.03%. Conclusions: H3.3 IHC could detect H3F3A G34W mutation in GCTB, but not for other rare mutation and wild types loci.

[Reassessment of EWSR1 gene rearrangement-negative undifferentiated round cell sarcomas in bone and soft tissues by fluorescence in situ hybridization].

Objective: To unravel the CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas from EWSR1 rearrangement-negative undifferentiated round cell sarcomas in the bone and soft tissues. Methods: Twenty-eight cases of EWSR1 rearrangement-negative undifferentiated round cell sarcomas of bone and soft tissues, tested for CIC rearrangement and BCOR rearrangement by fluorescence in situ hybridization and related immunostaining were analyzed, and some of the BCOR rearrangement cases were verified by reverse transcription-polymerase chain reaction. Results: Five of 28 (17.9%) tested cases were positive for CIC rearrangement and six (21.4%) for BCOR rearrangement. Histopathologically, CIC rearrangement sarcomas comprised nodular aggregates of round to polygonal cells, containing hyperchromatic nuclei, prominent nucleoli and moderate cytoplasm, with focal variable necrosis and myxoid stroma. BCOR-CCNB3 sarcomas mostly comprised diffusely arranged, round to oval to short spindly cells with angulated nuclei, vesicular chromatin, inconspicuous nucleoli and interspersed vessels. Immunohistochemically, five of six BCOR-CCNB3 sarcomas showed CCNB3 immunostaining, which could be helpful for diagnosis. Two patients with CIC rearrangement sarcoma died of the diseases in seven months and twenty-two months. One patient with BCOR-CCNB3 sarcoma died of the diseases in forty-six months. Conclusions: Overall, 39.3% of the EWSR1 rearrangement-negative undifferentiated round cell sarcomas are CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas. Molecular testing is helpful for diagnosis.

[Primary intraosseous Rosai-Dorfman disease: a clinicopathological analysis of fourteen cases].

Objective: To investigate the clinicopathological characteristics, histogenesis, immunophenotypes and molecular genetic features of primary intraosseous Rosai-Dorfman disease (RDD) for improving diagnostic accuracy and differential diagnosis. Methods: This retrospective study included 14 RDD cases diagnosed from January 2009 to January 2019 at Beijing Jishuitan Hospital, China. The immunohistochemical staining for S-100, cyclin D1, CD1a and CD207 expression was analyzed. The BRAF V600E and KRAS mutation analyses were performed using the Scorpions amplification refractory mutation system (ARMS) fluorescence quantitative PCR. Results: There were 6 female and 8 male patients, aged from 2 to 64 years (mean 31.4 years). All of the 14 cases occurred in the bone without lymph node disease, while one patient developed additional lesions within vertebra and nasal cavity. Radiographically, the lesions were lytic with sclerotic margins. Histologically, the lesions percolated through the medullary cavity in an infiltrative fashion and alternating hyper- and hypo-cellular regions of histiocytic clusters (seen as alternating dark and light zones at low magnification). Large histiocytes also showed emperipolesis. Some cases had areas of fibrosis and dense lymphoplasmacytic infiltrates. There were vasculitis and an increased number of plasma cells in the cases involving multiple sites. One case showed concurrence of RDD and Langerhans cell histiocytosis(LCH) with inconspicuous increase of Langerhans histiocytes. Immunohistochemical staining showed that the large histiocytes were positive for S-100, CD68 and CD163 in all cases. The nuclear immunoreactivity for cyclin D1 was observed in 13 of the 14 cases. S-100, CD1a and CD207 were positive in the case with concurrence of RDD and LCH. ARMS-PCR results showed that BRAF V600E mutation was observed in the cases with concurrence of RDD and LCH, while there were no KRAS mutations (7/7). Follow-up information was available for 12 patients and ranged from 9 to 49 months. Three of the 12 patients experienced recurrences after the first surgery. Conclusions: Primary intraosseous RDD is rare, and its concurrence with LCH is a very rare phenomenon. Its clinical symptoms, imaging, and pathological manifestations need to be distinguished from other bone lesions. The molecular detection of BRAF V600E and the nuclear expression of cyclin D1 mutations can be used for the diagnosis and differential diagnosis of RDD.

[Long-term observation of morphological changes of the inner retinal after internal limiting membrane peeling in macular hole surgery].

Objective: To investigate the characteristics of morphological changes of inner retinal layer after internal limiting membrane peeling in macular hole surgery. Methods: Retrospective case study. Patients with idiopathic macular hole from 2015 to 2018 underwent vitrectomy+internal limiting membrane peeling (inverting)+ gas tamponade in China-Japan Friendship Hospital were investigated. A total of 19 eyes (17 patients) were enrolled, including 4 males (4 eyes) and 13 females (15 eyes). The average age was 62.74±5.25 years. Optical coherence tomography (OCT) was used to obtain the topographic maps of retinal thickness, the thickness of retinal ganglion cell complex and probability maps, and the Angio/en-face maps of macular retina. The characteristics of the morphological changes of the inner retinal were comprehensively analyzed. Results: Among 19 eyes, 9 eyes had internal limiting membrane peeling, 8 eyes had lotus-like internal limiting membrane inverting, and 2 eyes had the uper180 degrees internal limiting membrane inverting. The minimum diameter of macular hole was (543.06+220.17) µm and the maximum diameter was (947.18+319.12) µm. The follow-up time was (21.05+9.66) months, and the visual acuity was 0.45+0.35 at the last follow-up. In the 19 eyes, all the macular holes were closed postoperatively and dissociated optic nerve fiber layer appearance (DONFL) and concentric macular dark spots (CMDS) all showed, as well as mGCC thinning. The changes of CMDS and mGCC in 2 eyes in the uper180 degrees internal limiting membrane inverting group were mainly seen in the upper retina and in the remaining 17 eyes were seen diffused around the macula, which roughly corresponded to the extent of internal limiting membrane peeling. Two eyes showed clear decrease of retinal capillary density on Angio/map. Conclusions: Long-term morphological changes of the inner retinal after internal limiting membrane peeling in macular hole surgery are obvious. In addition to the appearance changes like DONFL and CMDS, the macular ganglion cell complex (mGCC) is also involved. (Chin J Ophthalmol, 2019, 55:747-756).

[Dedifferentiated chordoma of sacrococcygeal region: a clinicopathologic analysis and review of literature].

Objective: To study the clinicopathologic features of dedifferentiated chordoma. Methods: Four cases of dedifferentiated chordoma of sacrococcygeal region were collected at Beijing Jishuitan Hospital, from 2009 to 2014. HE and immunohistochemistry (EnVision method) were used to observe the clinical, radiological and histological features of dedifferentiated chordoma and to make the diagnosis and differential diagnosis. The literature was reviewed. Results: Four cases of dedifferentiated chordoma were all located in the sacrococcygeal region. The mean age at diagnosis was 57 years (range 49-64 years). There were 1 female and 3 males. Histologically, there were two components of conventional chordoma and dedifferentiated sarcoma with or without transitional area between them. The histology of dedifferentiated components includes undifferentiated sarcoma and fibrosarcoma. Immunohistochemical study showed that cytokeratin, epithelial membrane antigen, S-100, Brachyury were positive in the portion of chordoma while p53 were positive in the portion of undifferentiated sarcoma. INI1 and vimentin were both positive. Conclusions: The dedifferentiated chordoma is very rare. The diagnosis should combine the histological characters and immunohistochemical results and should be differentiated from the sarcomatoid chordoma, poorly differentiated chordoma and other high grade sarcoma. p53 may play a role in the malignant transformation mechanism of chordoma.

[Dedifferentiated liposarcoma of extremities: a clinicopathologic analysis].

Objective: To study the clinicopathologic features of dedifferentiated liposarcoma of extremities. Methods: Nine cases of dedifferentiated liposarcoma of extremities diagnosed at Beijing Jishuitan Hospital from 2009 to 2017 were selected. The histological features of cases of dedifferentiated liposarcoma of extremities were evaluated by HE and immunohistochemistry, together with the clinical and radiological features. Flourescence in situ hybridization(FISH) was used to detect MDM2 amplification. Results: They were located in the thigh (6 cases), calf (2 cases) and buttock (1 case). There were six females and three males. Patients' age ranged from 61 to 79 years (mean 68 years). Histologically, there were two components, well-differentiated liposarcoma and dedifferentiated sarcoma with or without transitional lesions between them. The histology of dedifferentiated liposarcoma included undifferentiated sarcoma and fibrosarcoma. Heterologous elements such as bone and cartilage were present in two cases. Immunohistochemical study showed the tumor cells expressed vimentin, CDK4 and p16. MDM2 were positive in 6 cases (6/9) and p53 was positive in one case(1/9). CKpan was positive in the epithelioid differentiation area. S-100 protein was positive in the well-differentiated liposarcoma component. FISH showed the amplification of MDM2(6/9). Conclusions: Dedifferentiated liposarcoma of extremities is very rare. The diagnosis should be combined with the histological characteristics and immunohistochemical results and differentiated from the other tumors and tumor-like lesions.

[Clinical observation on the evolution process of macular ganglion cell complex and peripapillary retinal nerve fiber layer of neuritis patients].

Objective: To discuss the dynamic changes and correlation of macular ganglion cell (mGCC) and peripapillary retinal nerve fiber layer (pRNFL) of neuritis patients through optical coherence tomography (OCT). Methods: A retrospective case series study. Eleven eyes from 11 patients who have been diagnosed with neuritis and received regular follow-up during June 2013 through June 2015 were included. The dynamic characteristic changes of mGCC and pRNFL over the course of disease, as well as their correlations between best visual acuity and visual field have been analyzed based on OCT measurements. Results: According to the observation on the 11 eyes of the 11 patients, patients showed symptoms of decreased vision, abnormal visual field, swollen mGCC and pRNFL with normal or increased thickness during the subnormal period. During the advance-separation period (around 3 weeks), the thickness of mGCC decreased resulting from atrophy while pRNFL was still swollen. Druing the late period (usually 6-8 weeks after onset of the disease), both mGCC and pRNFL were getting thinner due to atrophy. Vision acuity and visual field of the patients improved after treatment, however, they were still not comparable with the normal level before the disease. Conclusions: The thickness changes of mGCC and pRNFL differs in neuritis patients over time. OCT can help us in detecting these changes, thus provide a foundation for us to further explore the treatment and anatomical changes of neuritis patients. (Chin J Ophthalmol, 2018, 54: 62-68).

Two-step identification of taro (Colocasia esculenta cv. Xinmaoyu) using specific psbE-petL and simple sequence repeat-sequence characterized amplified regions (SSR-SCAR) markers.

Colocasia esculenta cv. Xinmaoyu is an eddoe-type taro cultivar local to Taicang, Jiangsu Province, China; it is characterized by its pure flavor, glutinous texture, and high nutritional value. Due to its excellent qualities, the Trademark Office of the State Administration for Industry and Commerce of the People's Republic of China awarded Xinmaoyu, a geographical indication certification in 2014. Therefore, there is an urgent need to develop an efficient molecular marker for the specific identification of this cultivar, which would greatly facilitate the conservation and utilization of this unique germplasm resource. In the present study, amplifying the psbE-petL fragment from two dasheen-type and seven eddoe-type taro cultivars revealed three conserved insertions/deletions among sequences from the two taro types. Based on these sequence differences, a pair of site-specific primers was designed targeting the psbE-petL sequence from the dasheen-type taro, which specifically amplified a DNA band in all individuals from cultivars of this type, but not in those from the seven eddoe-type cultivars. To discriminate Xinmaoyu from the other eddoe-type taro cultivars, a pair of simple sequence repeat-sequence characterized amplified region (SSR-SCAR) primers was further developed to specifically amplify a DNA band from all Xinmaoyu individuals, but not from individuals of other eddoe-type taro cultivars. In conclusion, through a two-step-screening procedure using psbE-petL and SSR-SCAR markers, we developed a pair of primers that could specifically discriminate Xinmaoyu from nine taro cultivars commonly cultivated in Jiangsu Province and Fujian Province.

Analysis of genetic diversity of Brassica rapa var. chinensis using ISSR markers and development of SCAR marker specific for Fragrant Bok Choy, a product of geographic indication.

Non-heading Chinese cabbage [Brassica rapa var. chinensis (Linnaeus) Kitamura] is a popular vegetable and is also used as a medicinal plant in traditional Chinese medicine. Fragrant Bok Choy is a unique accession of non-heading Chinese cabbage and a product of geographic indication certified by the Ministry of Agriculture of China, which is noted for its rich aromatic flavor. However, transitional and overlapping morphological traits can make it difficult to distinguish this accession from other non-heading Chinese cabbages. This study aimed to develop a molecular method for efficient identification of Fragrant Bok Choy. Genetic diversity analysis, based on inter-simple sequence repeat molecular markers, was conducted for 11 non-heading Chinese cabbage accessions grown in the Yangtze River Delta region. Genetic similarity coefficients between the 11 accessions ranged from 0.5455 to 0.8961, and the genetic distance ranged from 0.0755 to 0.4475. Cluster analysis divided the 11 accessions into two major groups. The primer ISSR-840 amplified a fragment specific for Fragrant Bok Choy. A pair of specific sequence-characterized amplified region (SCAR) primers based on this fragment amplified a target band in Fragrant Bok Choy individuals, but no band was detected in individuals of other accessions. In conclusion, this study has developed an efficient strategy for authentication of Fragrant Bok Choy. The SCAR marker described here will facilitate the conservation and utilization of this unique non-heading Chinese cabbage germplasm resource.