Effect of internal phase particle size on properties of site mixed emulsion explosive at plateau environment.

To study the effect of internal particle size on the microstructure properties and thermal decomposition characteristics of site mixed emulsion explosive at different altitudes. Site mixed emulsion explosive was prepared with different shear rate. The particle size, viscosity, sensitized bubbles, detonation velocity and peak pressure of the emulsion explosive were tested after stored at different simulated altitudes. The thermal decomposition characteristics of emulsion matrix prepared at three different rotational speeds were measured by thermogravimetric analyzer and kinetic analysis was performed by non-isothermal model Kissinger-Akah-Sunose (KAS) method. The results show that with the increase in altitude, the internal phase size showed a trend of first increasing and then decreasing, and the number of sensitized bubbles within the emulsion explosive decreases. At an altitude of 0 m, the detonation velocity and peak overpressure of the emulsion explosive prepared by 1600 r min-1 increased 4.78% and 29.09%, respectively compared with 1200 r min-1, and at an altitude of 4500 m, the detonation velocity increased 11.87%, the peak overpressure increased 43.98%. The thermal decomposition activation energy of the emulsion matrix at 1600 r min-1 increased 13.14% compared to 1200 r min-1. It shows that in the production of site mixed emulsion explosive at high altitude, reducing the particle size of the internal phase of emulsion explosives in a certain range can effectively improve the performance of emulsion explosives.

[Effect of renal function on sarcopenia in elderly male patients with chronic kidney disease].

Objective: To evaluate the effect of renal function on sarcopenia in elderly male patients with chronic kidney disease (CKD). Methods: A total of 105 male CKD patients aged ≥65 years who were admitted to the Chinese PLA General Hospital between October 1, 2018 and January 30, 2019 were included in this study. Using two different equations to estimate glomerular filtration rate (GFR), respectively. According to the sarcopenia criteria, the participants were categorized as the non-sarcopenia group (n=72) and the sarcopenia group (n=33), respectively. The association of estimated GFR (eGFR) and the sarcopenia in the male CKD patients was analyzed using the model of multivariate logistic regression. Results: Among the 105 patients, the median age was 74 (68, 77) years old. The prevalence of sarcopenia was 31.4% (33/105). According to the multivariate logistic regression analysis, eGFR based on serum creatinine and Cys-C (eGFRscr-cys) lower than 45 ml·min(-1)·(1.73 m(2))(-1) (OR=4.17, 95%CI:1.08-16.02, P=0.038) and eGFR based on Cys-C (eGFRcys) lower than 45 ml·min(-1)·(1.73 m(2))(-1) (OR=3.99, 95%CI:1.08-14.75, P=0.038) were independent risk factors for underlying sarcopenic, respectively. The area under the receiver operating characteristics curve (AUC) revealed that eGFRscr-cys (AUC=0.67) was more suitable than eGFRcys (AUC=0.64) to predict the sarcopenia in elderly male patients with CKD. Conclusion: The increased incidence of sarcopenia in elderly men with CKD is accompanied with deterioration of renal function.

[Safety and efficacy of CD19-targeted CAR-T cells in 14 patients with refractory/relapsed Philadelphia chromosome-positive acute B-precursor lymphoblastic leukemia].

Objective: This study aimed to examine the safety and efficacy of CD19 chimeric antigen receptor T cell (CD19 CAR-T) therapy in relapsed/refractory Philadelphia chromosome-positive acute B-precursor lymphoblastic leukemia (R/R Ph(+) B-ALL) . Methods: The clinical data of 14 patients with R/R Ph(+) B-ALL treated with CD19 CAR-T cell therapy from November 2016 to April 2019 were retrospectively analyzed. Results: Among the 14 patients in this study, 7 were male and 7 were female, with a median age of 33 (7-66) years old. The efficacy was evaluated on the 28th day following CAR-T cells infusion; the overall response rate was 100.0% (14/14) , the complete response (CR) rate was 92.9% (13/14) , and the partial response (PR) rate was 7.1% (1/14) . After CAR-T cells infusion,12 cases (85.7%) developed cytokine release syndrome (CRS) : 1 case of grade 1 CRS, 4 cases of grade 2 CRS, 6 cases of grade 3 CRS, and 1 case of grade 4 CRS. Moreover, one case developed CAR T-cell-related encephalopathy syndrome (CRES) ; 14 cases had Ⅲ-Ⅳ hematological toxicity; and 13 CR cases had B cell dysplasia. These adverse reactions were all controllable. The median follow-up time was 441 (182-923) d. The median overall survival (OS) and progression-free survival (PFS) were 515 [95% confidence interval (CI) 287-743] days and 207 (95% CI 123-301) days, respectively. Conclusion: CD19 CAR-T cell therapy is safe and effective for R/R Ph(+) B-ALL treatment. However, the long-term efficacy needs to be further improved.

[Related factors of frailty in the elderly male patients with chronic kidney disease].

Objective: To investigate the characteristics of frailty in the elderly male patients with chronic kidney disease (CKD) and the effects of renal function on the incidence of frailty. Methods: A total of 105 non-dialysis CKD patients aged ≥65 years who were admitted to the Chinese PLA General Hospital between October 1, 2018 and January 30, 2019 were included in this study. Their clinical data and laboratory indicators were collected. Frailty was defined according to Fried frailty criteria. According to the frailty scores, the participants were categorized as non-frail (n=37), intermediately frail (n=37) and frail (n=31). The association of frailty and the level of estimated glomerular filtration rate (eGFR) in the patients was analyzed using the model of multivariate Logistic regression. Results: Among the 105 patients, the mean age was 74 (68, 77) years old. The incidence of frail and intermediate frail was 35.2% (37/105) and 29.5% (31/105), respectively. Multivariate logistic analysis showed statistically significant associations of frailty with age (OR=1.14, 95%CI:1.08-1.20, P<0.001), body mass index (OR=0.87, 95%CI:0.79-0.95, P=0.001) and the level of eGFR (OR=0.98, 95%CI:0.96-0.99, P=0.003) in those patients. The incidence of frail in patients with eGFR<45 ml·min(-1)·(1.73 m(2))(-1) and 45-59 ml·min(-1)·(1.73 m(2))(-1) was 1.02 (OR=2.02, 95%CI: 1.06~3.87) and 0.84 (OR=1.84, 95%CI: 1.05-3.22) times higher than that of eGFR≥60 ml·min(-1)·(1.73 m(2))(-1), respectively. Conclusion: The incidence of frailty in the elderly patients with CKD is affected by many factors, such as age, body mass index and renal function, and increases with decreased renal function.

[Efficacy and mechanism of loading dose clopidogrel in patients with transient ischemic attack and minor stroke].

Objective: To assess outcome, safety and possible mechanism of loading dose clopidogrel in patients with transient ischemic attack (TIA) and minor stroke. Methods: We reviewed patients with confirmed TIA and minor stroke admitted between July 2016 and December 2017 into the First Affiliated Hospital of Soochow University. Loss-of-function allele carriers of CYP2C19 were included and randomly divided into loading dose group (first dose of 300 mg clopidogrel) and standard dose group (first dose of 75 mg clopidogrel), 100 mg aspirin was gave at the same time, followed by aspirin 100 mg/d plus clopidogrel 75 mg/d maintaining for 20 days. Platelet aggregation (maximum aggregation ratio, MAR) induced by Adenosine diphosphate (ADP) was examined before and 3 days after administration. The National Institutes of Health Stroke Scale (NIHSS) score method was employed to assess the NIHSS scores before and after treatment in each group of patients; the modified Rankin Scale (mRS) was used to assess the 3-month functional outcome. Results: There was no significant difference in baseline data between the two groups (P>0.05).The proportion of early neurological function improvement in the two groups was 75.0% and 54.8%, and the difference was statistically significant (χ(2)=4.498, P=0.034). The 3-month prognosis was 79.5% and 61.3%, and the difference was statistically significant (χ(2)=4.000, P=0.045). Adverse events: 1 case in the loading dose group, 1 case in the standard dose group, the difference was not statistically significant (2.3% vs 1.6%, χ(2)=0.061, P=0.806). After 3 days of antiplatelet therapy, the MAR of the loading dose group decreased (11%±8%), and the MAR of the standard dose group decreased (9%±4%), the difference was statistically significant (P=0.013).In the loading dose group, there were 32 (72.7%)CYP2C19*2 carriers and 42 (95.5%)CYP2C19*2+*3 carriers; early neurological function improvement in 33 cases, accounting for 93.8% and 76.2%, respectively, and the difference was statistically significant (χ(2)=4.122, P=0.042). There were 35 patients with good prognosis in 3 months, accounting for 96.9% and 81.0%, respectively. The difference was statistically significant (χ(2)=4.310, P=0.038); MAR of CYP2C19*2 carrier was decreased (15%±5%), and MAR of CYP2C19*2+*3 carrier was decreased (12%±8%). The difference was statistically significant (P=0.039). Conclusions: Loading dose clopidogrel can improve the clinical prognosis of minor stroke/TIA without increasing the risk of bleeding. Loading dose clopidogrel may improve the prognosis of minor stroke/TIA by decreasing MAR of CYP2C19*2 carriers.

[A multi-center cross-sectional survey of medicine application in patients with osteoarthritis in China].

OBJECTIVE: To investigate the clinical characteristics, the medicine application and to evaluate the disease activity in patients with osteoarthritis (OA) in China. METHODS: This was a cross-sectional study. Totally 1 066 cases of OA from 40 hospitals in China from April to October 2017 were retrospectively enrolled. Demographic characteristics, clinical data, medicine application, and joint function were evaluated. All the data were analyzed by SPSS software 19.0. t test, Mann-Whitney U test and chi-square test were used for statistical analysis. RESULTS: In the 1 066 cases, the male-to-female ratio was 1:3.6 and the average age was (61.9±11.0) years, with an age range from 36 to 94 years. The incidence of knee OA, hip OA, and hand OA were respectively 81.9% (873/1 066), 14.1% (150/1 066), and 36.3% (387/1 066). In the study, 242 (22.7%) cases had two kinds of joint areas involved and three joint areas were involved in 51 cases (4.8%), and 56.6% (603/1 066) of the patients used more than one kind of non-steroid anti-inflammatory drugs (NSAIDs) while 61.2% (652/1 066) used disease modifying osteoarthritis drugs (DMOADs), including glucosamine (37.5%, 400/1 066), chondroitin sulfate (2.0%, 21/1 066), diacetate (5.9%, 63/1 066), and the combination of these drugs (15.8%, 168/1 066). 8.6% (92/1 066) patients only took analgesics to relieve the pain, not using any kind of NSAIDs or DMOADs. And 232 patients (21.7%) had intra-articular injections, including 9.2% (98/1 066) sodium hyaluronate, 4.5%(48/1 066) glucocorticoid, and 8.1% (86/1 066) combination of the two drugs. The proportion of the patients taking topical drugs accounted for 26.5% (283/1 066) and physical therapy accounted for 15.8% (168/1 066). Compared with those who suffered from knee OA, the patients who suffered from hip OA had more severe disease assessment. Moreover, there were significant differences in pain (Z=-7.625, P<0.001), morning stiffness (Z=-6.229, P<0.001), and joint function (Z=-6.777, P<0.001) between the two groups of the patients who suffered from knee or hip OA with The Western Ontario and McMaster Universities (WOMAC) osteoarthritis index. Furthermore, patients with hip OA took more analgesics (χ2=24.838, P<0.001). CONCLUSION: Oral NSAIDs and DMOADs are wildly used in patients with OA in China. However, the treatment of some patients still need to be improved. Patients with hip OA are more seriously ill and require aggressive treatment.

Expression of sodium/iodide transporters and thyroid stimulating hormone receptors in thyroid cancer patients and its correlation with iodine nutrition status and pathology.

OBJECTIVE: To investigate the relationship between the expression of sodium/iodide transporter (NIS) and thyroid stimulating hormone receptor (TSHR) and iodine nutritional status in patients with thyroid carcinoma. PATIENTS AND METHODS: 146 cases of thyroid cancer in Shanghai Gongli Hospital, the Second Military Medical University between February and December 2014 were selected as thyroid cancer group, 120 cases of normal thyroid morphology examined by thyroid ultrasound at the same period were selected as normal group. General information and thyroid function of two groups were recorded and analyzed. H&E staining was used to perform histopathological study on both normal group and thyroid cancer group, and immunohistochemistry was used to detect NIS and TSHR protein expression and position. Reverse transcriptase polymerase chain reaction (RT-PCR) was used for quantitative detection of NIS and TSHR mRNA in the two groups, and the relationship between iodine nutrition and NIS and TSHR expression in thyroid cancer patients was studied. The expression of NIS and TSHR in each group was detected by Western blotting, and the difference in NIS and TSHR expression was analyzed by SPSS 17.0 statistical software. RESULTS: The difference of serum total triiodothyronine (TT3), total thyroxine (TT4), and thyroid stimulating hormone (TSH) levels between the normal group and the thyroid cancer group was statistically significant (p < 0.05). H&E staining showed that the histopathology of the thyroid cancer group was significantly different from that of the normal group. Immunohistochemistry showed the positive expression of NIS and TSHR in thyroid cancer group. The expression of NIS and TSHR mRNA and protein in thyroid cancer patients was significantly lower than that in normal group detected by RT-PCR and Western blotting. Analysis of variance showed that the difference of NIS and TSHR expression between the two groups was statistically significant (p < 0.05). CONCLUSIONS: These findings indicated that the expression of NIS and TSHR in thyroid cancer is closely related to iodine nutritional status, which has important research value.

Correction to: Recommendations for the Development and Validation of Neutralizing Antibody Assays in Support of Biosimilar Assessment.

In the published article, the author B. Babbitt was cited as affiliation 9, but should have been cited as affiliation 2. In addition, there are 2 errors in the affiliations. The correct affiliations are shown in this erratum.

Recommendations for the Development and Validation of Neutralizing Antibody Assays in Support of Biosimilar Assessment.

The American Association of Pharmaceutical Scientists (AAPS) biosimilar focus group on nonclinical and clinical assays has developed this manuscript to guide the industry on best practices and testing strategies when developing neutralizing antibody (NAb) assays for biosimilar programs. The immunogenicity assessment to biosimilar and originator drug products is one of the key aspects of clinical programs for biosimilars to demonstrate biosimilarity. Establishing that there are no clinically meaningful differences in immune response between a proposed product and the originator product is a key element in the demonstration of biosimilarity. It is critical to collect, evaluate, and compare the safety and immunogenicity data from the clinical pharmacology, safety, and/or efficacy studies especially when the originator drug product is known to have potential for immune-mediated toxicity. This manuscript aims to provide a comprehensive review and recommendations on assay formats, critical reagents, approaches to method development, and validation of the neutralizing antibody assays in extrapolation within the scope of biosimilar drug development programs. Even if there are multiple options on the development and validation of NAb assays for biosimilar programs, the type of drug and its MoA will help determine the assay format and technical platform for NAb assessment (e.g., cell-based or non-cell-based assay). We recommend to always perform a one-assay approach as it is better to confirm the biosimilarity using one-assay for NAb. If a one-assay approach is not feasible, then a two-assay format may be used. This manuscript will provide all the details necessary to develop NAb assays for biosimilars.

[Effect of interleukin-6 promoter DNA methylation on the pathogenesis of systemic lupus erythematosus].

Objective: To analyze peripheral blood interleukin-6 (IL-6) promoter DNA methylation status and its clinical significance in patients with systemic lupus erythematosus (SLE). Methods: Blood samples of 41 adult patients with SLE and 20 healthy controls were collected.The methylation status of IL-6 promoter was determined by methylation specific polymerase chain reaction (MSP). The IL-6 expression was detected by real-time PCR.Correlations between IL-6 promoter methylation status and clinical features or laboratory findings in patients with SLE were analyzed. Results: The levels of IL-6 mRNA were significantly higher in peripheral blood of SLE.DNA methylation levels of IL-6 promoter were reduced in SLE patients as compared with healthy controls.The methylation status and expression of IL-6 in peripheral blood reflected the levels in peripheral blood mononuclear cells (PBMCs). Significantly positive correlation was found between IL-6 hypomethylation and renal disorder, as well as hypocomplementemia in patients with SLE. Conclusion: Hypomethylation of interleukin-6 promoter in peripheral blood might be involved in the etiology of SLE.

[A cross-sectional study on application of glucocorticoid in systemic lupus erythematosus patients].

Objective: To explore the status of glucocorticoid application in patients with systemic lupus erythematosus (SLE) in China. Methods: Epidemiological survey was used. The SLE patients who met the 1997 classification criteria of American College of Rheumatology were enrolled. The usage of glucocorticoid and related adverse reactions were recorded and analyzed. Results: A total of 400 SLE patients were enrolled, including 35 men and 365 women. The average age was (37.4±14.0) years old, and the average duration of disease was (6.7±5.8) years. There were 310 patients using glucocorticoid as maintenance. Sixty-one percent (n=244) patients started using medium dose (prednisone 30-<60 mg/d) as the initial treatment of glucocorticoid, which lasted for(37±11)days.The time of drug duration in patients with low dose prednisone (7.5-<30 mg/d)and high dose (60-100 mg/d) was(92±20)and(17±3)days respectively (P<0.05 between 3 groups). However, patients receiving different initial dosage were of no discrepancy in the maintenance therapy. During maintenance, even though 51.0% (n=158) patients were on prednisone 2.5-5 mg/d, the duration of drug use in >5-10 mg/d groupwas longer[(29.9±3.3) months]. Patients with involvement of internal organs had a higher tendency to use 60-100 mg/d prednisone or pulse-dose therapy in the initial treatment, nevertheless these two groups had no difference of maintenance dosage. Among all 400 patients, 62 patients withdrew glucocorticoid, including 17 patients with disease remission (4.3%), 44 by self-withdrawal and one with adverse reaction. Conclusion: In China, the medium dosage of glucocorticoid is the most common initial treatment in patients with SLE.Prednisone 2.5-5 mg/d was the most common choicefor maintenance therapy. Currently, the proportion of glucocorticoid withdrawal remains low in SLE patients achieving remission.

Collinearity analysis of allotetraploid Gossypium tomentosum and Gossypium darwinii.

Gossypium tomentosum and G. darwinii are wild allotetraploid cotton species, characterized by many excellent traits, including fiber fineness, drought tolerance, and Fusarium and Verticillium wilt resistance. Based on the construction of F2 linkage groups of G. hirsutum x G. tomentosum and G. hirsutum x G. darwinii, two genetic linkage maps were compared. As a result, we found a total of seven inverted fragments on chr02, chr05, chr08, chr12, chr14, chr16, and chr25, and three translocated fragments on chr05, chr14, and chr26. In addition, comparison of the inverted and translocated fragments revealed that the orientation of four of seven markers in G. tomentosum were consistent with G. hirsutum or G. raimondii. The orientation of one of seven inverted markers of G. darwinii was consistent with G. hirsutum, and the orientation of one of three translocated markers of G. tomentosum was consistent with G. raimondii. These results indicate that, in comparison to G. darwinii, G. tomentosum has a closer genetic relationship to G. hirsutum. These findings will be important for our understanding on the genome structure of G. tomentosum and G. darwinii, and set the scene for further in-depth genome research such as fine mapping, tagging genes of interest from wild relatives, and evolutionary study.

Mapping QTLs for drought tolerance in an F2:3 population from an inter-specific cross between Gossypium tomentosum and Gossypium hirsutum.

Cotton is one of the most important natural fiber crops in the world. Its growth and yield is greatly limited by drought. A quantitative trait locus (QTL) analysis was therefore conducted to investigate the genetic basis of drought tolerance in cotton (Gossypium spp) using 188 F2:3 lines developed from an inter-specific cross between a wild cotton species, G. tomentosum, and an upland cotton, G. hirsutum (CRI-12). A genetic map was constructed using 1295 simple sequence repeat markers, which amplified 1342 loci, distributed on 26 chromosomes, covering 3328.24 cM. A field experiment was conducted in two consecutive years (2014 and 2015) and 11 morphological and physiological traits were recorded under water-limited (W1)/well-watered (W2) regimes at three growth stages (bud, flowering, and full boll). The traits measured included chlorophyll content, plant height, leaf area, leaf number, leaf fresh weight, leaf dry weight, boll weight, number of bolls per plant, and the number of fruiting branches. Sixty-seven and 35 QTLs were found under the W1 and W2 conditions, respectively. Of these, the majority exhibited partial dominance or over-dominance genetic effects for increasing the trait values. Four consistent QTLs were found under the W1 treatment on chromosomes 5, 8, 9, and 16, whereas no consistent QTL was found in W2. Thirteen QTL clusters were also identified on nine chromosomes (2, 3, 5, 6, 9, 14, 15, 16, and 21). These results will help to elucidate the genetic basis of drought tolerance in cotton.

[Pathogens and clinical characteristics of bacterial infection in hematology department between 2010 and 2014].

OBJECTIVE: To analyze the characteristics of distribution and drug resistance of bacterial infection in several different parts of hematology department inpatients of Anhui Provincial Hospital from January 2010 to December 2014, including patients who had received hematopoietic stem cell transplantation (HSCT). METHODS: Anti-microbial susceptibility test was done by Kirby-Bauer method and automated systems and the data were analysed by WHONET 5.6 software. RESULTS: A total of 3 312 copies of inspection samples were analyzed, including 2 716 (82%) blood samples and other 596 specimens (18%). 634 bacterial strains were isolated from 3 312 samples (19.14%) including 488 samples (76.97%) from blood culture. 427 (67.35%) bacterial strains were gram-negative, and the other 207 (32.65%) were gram-positive. Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were most common gram-negative bacterial and the resistant rates to imipenem were 0.8%, 11.8% and 3.3%, respectively. Detection rates of Extended-spectrum beta-lactamases in Escherichia coli and Klebsiella pneumoniae were 83.9% and 75.0%, respectively. At the same time, Coagulase negative Staphylococcus, Streptococcus and Enterococcus were most common kinds of gram-positive bacteria. Methicillin-resistant coagulase negative staphylococcus accounted for 65.9% antibiotic resistance. No vancomycin and/or linezolid and/or tigecycline resistant strains of Staphylococcus spp. and Enterococcus spp. were found in those patients. CONCLUSION: Patients with hematology diseases had a higher risk of bacterial infections, mainly caused by Gram-negative bacteria. There are different distributions of bacterial in different wards.

Radiological follow-up results of untreated anterior disc displacement without reduction in adults.

The aim of this study was to assess the radiographic changes in untreated adults with bilateral anterior disc displacement without reduction. A cohort study was designed to compare the bone changes, effusion, disc configuration, and pseudo-disc changes on two magnetic resonance images obtained at least 24 months apart. Twenty-eight patients (22 female, six male) with a mean age of 33.1 years (range 20-57 years) were included. The mean interval between the initial visit and the follow-up visit was 36.2 months. At the initial visit, the frequencies of bone changes, effusion, disc deformation, and pseudo-disc changes were 51.79%, 35.71%, 100%, and 0%, respectively. At follow-up, the frequency of effusion had decreased significantly. The frequency of bone changes had increased significantly to 75%, but newly formed cortical bone was present in five condyles. All discs remained deformed. Pseudo-disc changes were detected in five joints. Over a long period of observation, there was a significant decrease in effusion and a significant increase in bone changes. However, some adaptive changes occurred.

Construction and characterization of a bacterial artificial chromosome library for the allotetraploid Gossypium tomentosum.

Gossypium tomentosum is a wild allotetraploid species with the (AD)5 genome. It is characterized by many useful traits including finer fiber fineness, drought tolerance, and Fusarium and Verticillium resistance. We constructed the first bacterial artificial chromosome library for Gossypium tomentosum. With high quality and broad coverage, this library includes 200,832 clones, with an average insert size of about 122 kb and fewer than 3% empty clones. Our library is approximately 10-fold the size of the (AD)5-genome (2400 Mb) and provides a 99.7% probability of isolating genes of interest or their sequences. Seven of eight simple sequence repeats markers that are located on five different chromosomes and linked with resistance to Verticillium wilt could amplify the 50 superpools and obtained one to five hits. This high capacity library will be an important genomic resource for classifying and analyzing the evolution of allotetraploid cotton species as well as for isolating disease-resistance and drought-tolerance genes.

Genetic diversity and relationship analysis of Gossypium arboreum accessions.

Simple sequence repeat techniques were used to identify the genetic diversity of 101 Gossypium arboreum accessions collected from India, Vietnam, and the southwest of China (Guizhou, Guangxi, and Yunnan provinces). Twenty-six pairs of SSR primers produced a total of 103 polymorphic loci with an average of 3.96 polymorphic loci per primer. The average of the effective number of alleles, Nei's gene diversity, and Shannon's information index were 0.59, 0.2835, and 0.4361, respectively. The diversity varied among different geographic regions. The result of principal component analysis was consistent with that of unweighted pair group method with arithmetic mean clustering analysis. The 101 G. arboreum accessions were clustered into 2 groups.