[Establishment and validation of a predictive model for neurogenic urinary tract injury in children].

Objective: To establish a predictive model for upper urinary tract damage in children with neurogenic bladder and verify its efficacy. Methods: From January 2011 to December 2021, 143 children with NB in the Children's Hospital of Chongqing Medical University and 84 children with NB in the First Affiliated Hospital of Zhengzhou University were selected as the research objects. The former is set as the training set and the latter is set as the validation set, and the general parameters of the two are compared. The independent risk factors of upper urinary tract damage in children with NB were screened out by Lasso regression, and multivariate logistic regression analysis and a nomogram prediction model was established. The models were validated internally and externally on the training set and validation set, respectively, and the area under the receiver operating curve (ROC) was used to verify the accuracy of the model. Results: A total of 227 children with NB were included in this study, including 121 males and 106 females, aged (10.2±3.8) years. There was no significant difference in other parameters except age between the training set and validation set (all P>0.05); Lasso regression and multivariate logistic regression analysis showed that detrusor leakage point pressure (DLPP) ≥ 40 cmH2O (OR=4.76, 95%CI: 2.01-11.26, 1 cmH2O=0.098 kPa), overactive bladder (OAB) (OR=3.08, 95%CI: 1.34-7.04), bladder compliance (BC)<20 ml/cm H2O (OR=3.65, 95%CI: 1.41-9.47), history of previous urinary tract infection (OR=2.73, 95%CI: 1.09-6.81), and abdominal pressure/other voiding patterns (OR=2.86, 95%CI: 1.20-6.82) were risk factors for upper urinary tract damage in children with NB (all P<0.05). The above parameters were used to establish a nomogram model of upper urinary tract damage in children with NB. The internal and external validation results show that the AUC values for the training and validation sets were 0.84 (95%CI: 0.77-0.91) and 0.86 (95%CI: 0.79-0.94), respectively. Conclusion: The prediction model of upper urinary tract damage in children with NB constructed in this study has high discrimination, accuracy and clinical applicability, which can help clinicians identify high-risk patients and make individualized treatment design for these patients.

[A comparative study of the effects of different treatment strategies on postoperative anal function and quality of life in patients with complete obstructive left hemicolon cancer].

Objective: To compare the effects of 3 treatment strategies (emergent surgery, self-expanding metallic stents, self-expanding metallic stents plus neoadjuvant chemotherapy) on postoperative anal function and quality of life in patients with complete obstructive left hemicolon cancer. Methods: A retrospective cohort study was conducted. Clinical data of patients with complete obstructive left hemicolon cancer admitted to General Surgery Department of Beijing Chaoyang Hospital between January 2017 and October 2019 were retrospectively collected. Patient inclusion criteria: (1) complete obstructive left hemicolon cancer was confirmed through clinical manifestation and abdominal computed tomography; (2) adenocarcinoma was confirmed by postoperative pathology; (3) emergent radical resection of primary tumor was performed with temporary stoma, or radical resection of primary tumor and primary anastomosis was performed without stoma, 7 to 14 days after completion of insertion of self-expanding metallic stents. Patients who did not receive stoma reversion after emergent operation were excluded. According to different therapies, patients were divided into three groups: emergent surgery (ES) group, self-expanding metallic stents (SEMS) group and self-expanding metallic stents plus neoadjuvant chemotherapy (SEMS+NAC) group. Wexner score for incotinence (higher score indicates the worse anal function), Vaizey score (>10 indicates fecal incontinence) and low anterior resection syndrome (LARS) scale (higher score indicates the worse anal function) were applied to evaluate anal function of patients among groups at postoperative 1-, 6- and 12-month. EORTC QLQ-C30 questionnaire was used to assess the quality of life. Risk factors of decreased anal function were identified by logistic regression analysis. Results: A total of 72 patients were enrolled, including 27 (37.5%) patients in ES group, 23 (31.9%) in SEMS group and 22 (30.6%) in SEME+NAC group. The baseline characteristics including age, gender, tumor location, comorbidities, total blood loss, operation time and postoperative complications, were comparable among groups, except that the proportion of laparoscopic surgery was significantly lower in ES group (4/27, 14.9%) than that in SEMS (15/23, 65.2%) and SEMS+NAC group (16/22, 72.7%) with significant difference (P<0.001). The follow-up ended up to October 2020, and the overall follow-up rate was 79.2% (57/72). No significant differences existed in the Wexner score of patients among groups at postoperative 1-, 6- and 12-month (all P>0.05). The Vaizey scores at postoperative 1-month in ES, SEMS and SEMS+NAC group were 7 (0-17), 3 (0-7) and 4 (0-8) respectively with significant difference (H=18.415, P=0.001), and the scores in SEMS and SEMS+NAC groups were significantly lower than that in ES group (both P<0.05), while no significant difference existed between SEMS and SEMS+NAC group (P>0.05). Vaizey scores at postoperative 6- and 12-month among 3 groups were not significantly different (both P>0.05). The LARS scores at postoperative 1-month in ES, SEMS and SEMS+NAC groups were 20 (0-37), 15 (0-24) and 16 (0-28) respectively with significant difference (H=3.660, P=0.036), and the scores in SEMS and SEMS+NAC groups were significantly lower than that in ES group (both P<0.05), while no significant difference existed between SEMS and SEMS+NAC groups (P>0.05). LARS scores at postoperative 6- and 12-month among 3 groups were not significantly different (both P>0.05). The QLQ-C30 score revealed that the social function of patients in SEMS group and SEMS+NAC group was significantly better than that in ES group (both P<0.05), while no significant difference existed between SEMS and SEMS+NAC group (P>0.05). The logistic regression analysis revealed that only ES was an independent risk factor of decreased anal function (OR=2.264, 95% CI: 1.098-4.667, P=0.027). Conclusion: Compared to ES, SEMS may improve quality of life and short-term anal function of patients with complete obstructive left hemicolon cancer.

LncRNA CASC19 contributed to the progression of pancreatic cancer through modulating miR-148b/E2F7 axis.

OBJECTIVE: Cancer susceptibility 19 (CASC19), a crucial lncRNA associated with multiple cancers, has been reported to play a vital role in the progression of human malignant tumors. However, the underlying mechanism of CASC19 in pancreatic cancer (PC) was still unknown. The purpose of this study was to explore the biological and clinical significance of CASC19 in PC. PATIENTS AND METHODS: RT-qPCR assay was adopted to analyze CASC19 expression in PC tissues and cell lines. Furthermore, the correlation between the CASC19 level and the survival rate of PC patients was assessed by Kaplan-Meier analysis. Bioinformatics analysis and Luciferase reporter assay were utilized to confirm the interaction between miR-148b and CASC19 or E2F7. Cell viability, migration, invasion, and apoptosis were analyzed using MTT, transwell, and TUNEL assays. RESULTS: The results elucidated that CASC19 expression was markedly increased in PC tissues and cell lines. Patients with high expression of CASC19 had a short survival time. Silencing of CASC19 attenuated PC cell proliferation, migration, and invasion. Moreover, we identified that miR-148b was a target of CASC19. CASC19 was negatively correlated with miR-148b and positively correlated with E2F7. The inhibitory effect of CASC19 knockdown on the progression of PC was reversed by the down-regulation of miR-148b or up-regulation of E2F7. CONCLUSIONS: These results demonstrated that CASC19 participated in the development of PC. The CASC19/miR-148b/E2F7 axis might be a new study direction for PC treatment.

Testicular hypertrophy as predictor of contralateral nonpalpable testis among Chinese boys: An 18-year retrospective study.

OBJECTIVE: To assess the accuracy of contralateral testis hypertrophy for predicting the fate of nonpalpable testis in Chinese boys at different ages. METHODS: The data of patients who presented with unilateral impalpable testis and who underwent laparoscopy at the Children's Hospital of Chongqing Medical University between January 1, 2000 and January 1, 2018 were reviewed. The boys were divided into four groups: age-matched volunteers with no testicular abnormalities represented the control group (group I), boys with palpable undescended testis (group II), boys with nonpalpable testis (NPT)/viable testis (VT) (group III), and boys with NPT/non-viable testis (NVT) group (group IV). Scrotal testes were prospectively measured by ultrasonography for volume and size, and diagnostic laparoscopy was performed to determine the state of the cryptorchid testis. RESULTS: The mean contralateral testicular volume and length in the boys with an absent testis was 0.78mL and 17mm compared with 0.67mL and 15mm in the boys with a testis present and 0.63mL and 15mm in the controls, respectively (P<0.05). The predictive accuracy, sensitivity, and specificity for an absent testis were 64.9%, 75%, and 49%, respectively, for volume and 64.2%, 56.3%, and 76.4%, respectively for length at the optimal cutoff value of 0.65mL volume and 16.55mm length. Contralateral testis volume was the most accurate in predicting monorchism in 0-2-year-olds (sensitivity: 75%, specificity: 70%, accuracy: 73.1%) and the contralateral testicular length was most accurate for 4-6-years-old (sensitivity: 68.6%; specificity: 77.8%; accuracy: 72.2%). We also included 29 patients with bilateral undescended testis (UDT) and with unilateral nonpalpable. Cutoff values for testicular volume and length were 0.6mL (sensitivity: 81.8%, specificity: 88.9%, accuracy: 86.2%) and 13.5mm (sensitivity: 63.6%, specificity: 77.8%, accuracy: 77.8%). CONCLUSION: The present results exclusively obtained from laparoscopic exploration suggest that a testis volume of>0.65mL or a testis length of>16.55mm could predict monarchism with an accuracy of about 65%. In younger patients aged 0-2 years and 4-6 years, the overall predictive accuracy increases to about 73% but laparoscopic exploration is still required.

[Comparison of short-term efficacy and perioperative safety between neoadjuvant therapy and total neoadjuvant therapy in patients with locally advanced rectal cancer].

Objective: To compare the short-term efficacy and perioperative safety of neoadjuvant chemoradiotherapy (nCRT) with total neoadjuvant treatment (TNT) in patients with locally advanced rectal cancer (LARC). Methods: A retrospective cohort analysis was carried out. Inclusion criteria: (1) rectal adenocarcinoma confirmed by pathology with a distance from tumor inferior border to anal verge within 12 cm; (2) clinical stage cT3-4N0 or cT1-4N1-2 diagnosed by magnetic resonance imaging (MRI) or endorectal ultrasonography; (3) a single rectal tumor confirmed by colonoscopy; (4) patients suitable for chemoradiotherapy; (5) no previous history of other tumors. Exclusion criteria: (1)patients with previous rectal cancer surgery and local recurrence; (2) those who did not complete nCRT course; (3) those with distant metastases; (4) those with defective clinicopathological data. According to the above criteria, a total of 134 LARC patients at the Department of General Surgery of Beijing Chaoyang Hospital from January 2016 to January 2019 were enrolled, including 82 males and 52 females, with a male-female ratio of 1.58∶1.00 and mean age of (59.6±11.2) (26-81) years. Based on neoadjuvant regimen, patients were divided into nCRT group (n=55) and TNT group (n=79). There were no statistically significant differences in baseline data, such as age, sex, distance from tumor to anal verge, Eastern Cooperative Oncology Group (ECOG) performance status and clinical TNM stage, between the two groups (all P>0.05). All the patients received pelvic intensity-modulated radiotherapy (IMRT) with a total dose of 50.4 Gy in 28 fractions. Patients in nCRT group received oral capecitabine chemotherapy during radiotherapy and underwent surgery 6-8 weeks after chemoradiation. Patients in TNT group received one cycle of induction CapeOX (oxaliplatin and capecitabine) and concurrent chemoradiotherapy, then underwent a radical surgery two weeks after completion of consolidation chemotherapy. The efficacy of neoadjuvant therapy, adverse events of chemoradiotherapy and perioperative safety were compared between the two groups. Results: Patients of two groups completed the course of neoadjuvant therapy. There were no statistically significant differences between nCRT group and TNT group in the incidence of adverse events in neutropenia [7.3% (4/55) vs. 10.1% (8/79)], anemia [3.6% (2/55) vs. 3.8% (3/79)], thrombocytopenia [5.5% (3/55) vs. 7.6% (6/79)], gastrointestinal dysfunction [3.6% (2/55) vs. 6.3% (5/79)] and radiation enteritis [9.1% (5/55) vs. 8.9% (7/79)] (all P>0.05). One hundred and thirty patients completed TME surgery, including 54 patients in nCRT group and 76 patients in the TNT group. Compared with the nCRT group, the proportion of abdominoperineal resection (APR) was higher in the TNT group [31.6% (25/76) vs. 13.0% (7/54), χ(2)=9.382, P=0.009]. No statistically significant differences in morbidity of postoperative complication, operation time, intraoperative blood loss and postoperative hospital stay between the two groups were found (all P>0.05). The distal and circumferential margins were negative in all the patients. Seventeen patients in the TNT group 22.4% (17/76) got pathologic complete response (pCR), which was significantly higher than 7.4% (4/54) in nCRT group (χ(2)=5.217, P=0.022). There were no statistically significant differences in ypTNM classification, perineural invasion and venous invasion between the two groups (all P>0.05). Conclusion: The pCR of TNT is higher than that of nCRT without increasing the incidence of toxicity and complications of radiotherapy and chemotherapy for patients with locally advanced rectal cancer.

Urine-derived stem cells for the therapy of diabetic nephropathy mouse model.

OBJECTIVE: Diabetic nephropathy (DN) is one of the most representative diabetic microangiopathy complications. So far, there have been no satisfactory therapeutic strategies, and the injection of stem cells provides a target for DN therapy. PATIENTS AND METHODS: Urine-derived stem cells (USCs) were obtained from 9 healthy men. 24 mice were randomly and equally divided into control group, DN model group, DN+hUSC group (treated with USCs for 3 times). Hematoxylin-eosin (HE) and Masson staining were used to detect histological changes of kidney injury. Creatinine and blood urea nitrogen (BUN) were measured to assess renal function. Besides, myofibroblast accumulation, macrophage infiltration, cell proliferation, and oxidative stress were detected by immunohistochemical analysis. RESULTS: Compared with DN model group, DN+hUSC group showed lower function loss, cell infiltration, and oxidative stress, as well as less renal fibrosis, histological damage, and cell proliferation. CONCLUSIONS: USC can alleviate inflammation and oxidative stress, reduce renal interstitial fibrosis, improve renal tissue structure and protect renal function through paracrine effect.

[Effect of endoscopic stent placement combined with neoadjuvant chemotherapy on short-term and long-term results in patients with acute left-sided malignant colorectal obstruction without distant metastases].

Objective: This study aimed to evaluate the outcomes of colorectal obstruction patients without distant metastases treated with different strategies. Methods: This retrospectively study included 82 patients who presented in Beijing Chaoyang Hospital from 2010 to 2015 with acute left-sided malignant colorectal obstruction. Patients with distant metastases were excluded. After informed consent, patients were divided into colonic stenting (SEMS group, n=28) , neoadjuvant chemotherapy(NCT group, n=15) or immediate emergency surgery(control group, n=39). Patients who had successful colonic stenting underwent elective surgery 1 to 2 weeks later or underwent neoadjuvant chemotherapy before elective surgery, while the other group had emergency surgery. Short-term data on postoperative mortality, morbidity, length of intensive care and hospital stay were compared. Overall survival and disease-free survival were also analyzed. Results: Patients in the three study arms had similar demographic profiles. The laparoscopic resection of the NCT and SEMS group was higher than that of the control group, the stoma rate was lower, and the differences were statistically significant[73.3% (11/15) , 42.9% (12/28) vs 25.6% (10/39) (P=0.006) and 13.3% (8/15) , 28.6% (8/28) vs 66.7% (26/39) (P<0.001) respectively].Compared with the SEMS and NCT group, the control group had a higher rate of postoperative complications, less of retrieved lymph nodes, longer of intensive care and lower total hospitalization expenses, and the difference was statistically significant[32.1% (9/28) , 13.3% (2/15) vs 59.0% (23/39) (P=0.004) , 21 (16,25) , 23 (19,34) vs 17 (13,25) (P=0.02) , 1.5 (0,3.0) , 1.0 (0,3.0) vs 3.0 (1.0, 4.0) (P=0.028) and 7.3 (2.8,14.1) , 11.1 (6.9,18.5) vs 7.1 (3.3,37.4) (P=0.004) respectively]. The overall and disease-free survival rate of the NCT group were higher than the SEMS group and control group, and the difference was statistically significant[93.3% (14/15) , 57.1% (16/28) vs 61.5% (24/39) (P=0.033) and 86.7% (13/15) , 53.6% (15/28) vs 51.3% (20/39) (P=0.047) respectively]. There was no significant difference among the NCT, SEMS and control group in the rate of systemic recurrence of the[6.7% (1/15) , 25.0% (7/28) vs 28.2% (11/39) (P=0.243) ]. Conclusions: For acute left-sided malignant colorectal obstruction without distant metastases, endoscopic stent placement combined with NCT not only is a bridge to elective operation, but also significantly improves the long-term results.

[Body mass index and related attribution to all-cause mortality in adults of Family Cohort of Nutrition and Chronic Diseases in Shanxi province].

Objective: To assess the relationship between body mass index (BMI) and mortality in adults of Shanxi, China. Methods: Baseline data were from the '2002 China Nutrition and Health Survey' in Shanxi province. All the death-related investigation and follow-up visits were carried out from December 2015 to March 2016. The follow-up program covered 5 360 people from all the 7 007 participants aged 18 years and over that having complete core information, with a rate as 76.5%. Participants of this study were divided into eight groups, according to the appearance of BMI. Taking the group with the lowest mortality density as the reference group, Cox regression model was used to estimate the hazard ratio (HR) and 95% confidence interval (CI) of deaths by the whole population, gender and age groups (≥60 years, <60 years). Results were then adjusted by age, gender, smoking, alcohol use and education level from the baseline survey. Sensitivity analysis was also conducted. Results: Results from the study showed that among the total number of 67 129 person- years from the average period of 12.5 years, there were 615 deaths occurred, with the mortality density as 916 per 100 000 person-years. Taking the BMI range of 26.0-27.9 kg/m(2) as the reference, the aHRs of death increased to 1.90 (95%CI: 1.26-2.86), 1.68 (95%CI: 1.15-2.45), 1.49 (95%CI: 1.08-2.06) and 1.72 (95%CI: 1.07-2.76) after the multivariate adjustment, in these four groups (BMI<18.5, 18.5-19.9, 22.0-23.9 and ≥30.0 kg/m(2)), respectively. Low body weight (BMI<18.5 kg/m(2)) was associated with higher risks of death in the elderly of ≥60 years, with the aHR of death as 1.94 (95%CI: 1.20-3.15). Conclusions: When BMI appeared as ≤19.9 kg/m(2), 22.0-23.9 kg/m(2) and ≥30.0 kg/m(2), the risks of death would increase. In addition to programs that focusing on obesity, special attention should be paid to the high risk of mortality which was caused by low-weight and malnutrition in the elderly.

lncRNA MEG3 inhibit proliferation and metastasis of gastric cancer via p53 signaling pathway.

OBJECTIVE: lncRNA MEG3 has been reported as a tumor suppressor gene in many different kinds of cancer, but its role in gastric cancer has not been fully understood. Then, we would like to explore its mechanism in gastric cancer. PATIENTS AND METHODS: We first used qRT-PCR to detect the expression of lncRNA MEG3, then CCK8 and wound healing assay were used to detect the effect of lncRNA MEG3 on gastric cancer cells. Western blot assay was used to measure the expression of p53 when lncRNA MEG3 was overexpressed. RESULTS: lncRNA MEG3 was highly expressed in the adjacent tissue, compared to the one in gastric cancer tissue. What's more, we also found that overexpression of lncRNA MEG3 could decrease the proliferation and metastasis of gastric cancer cells. Finally, overexpression of lncRNA MEG3 could also increase the expression of p53. CONCLUSIONS: lncRNA MEG3 could inhibit the proliferation and metastasis of gastric cancer.

Oncogenic K-Ras upregulates ITGA6 expression via FOSL1 to induce anoikis resistance and synergizes with αV-Class integrins to promote EMT.

In many cancer types, integrin-mediated signaling regulates proliferation, survival and invasion of tumorigenic cells. However, it is still unclear how integrins crosstalk with oncogenes to regulate tumorigenesis and metastasis. Here we show that oncogenic K-RasV12 upregulates α6-integrin expression in Madin-Darby canine kidney (MDCK) cells via activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK)/Fos-related antigen 1-signaling cascade. Activated α6-integrins promoted metastatic capacity and anoikis resistance, and led to perturbed growth of MDCK cysts. Transcriptomic analysis of K-RasV12-transformed MDCK cells also revealed robust downregulation of αV-class integrins. Re-expression of αV-integrin in K-RasV12-transformed MDCK cells synergistically upregulated the expression of Zinc finger E-box-binding homeobox 1 and Twist-related protein 1 and triggered epithelial-mesenchymal transition leading to induced cell motility and invasion. These results delineate the signaling cascades connecting oncogenic K-RasV12 with α6- and αV-integrin functions to modulate cancer cell survival and tumorigenesis, and reveal new possible strategies to target highly oncogenic K-RasV12 mutants.

Targeted re-sequencing of linkage region on 2q21 identifies a novel functional variant for hip and knee osteoarthritis.

OBJECTIVE: The aim of the study was to identify genetic variants predisposing to primary hip and knee osteoarthritis (OA) in a sample of Finnish families. METHODS: Genome wide analysis was performed using 15 independent families (279 individuals) originating from Central Finland identified as having multiple individuals with primary hip and/or knee OA. Targeted re-sequencing was performed for three samples from one 33-member, four-generation family contributing most significantly to the LOD score. In addition, exome sequencing was performed in three family members from the same family. RESULTS: Genome wide linkage analysis identified a susceptibility locus on chromosome 2q21 with a multipoint LOD score of 3.91. Targeted re-sequencing and subsequent linkage analysis revealed a susceptibility insertion variant rs11446594. It locates in a predicted strong enhancer element region with maximum LOD score 3.42 under dominant model of inheritance. Insertion creates a recognition sequence for ELF3 and HMGA1 transcription factors. Their DNA-binding affinity is highly increased in the presence of A-allele compared to wild type null allele. CONCLUSION: A potentially novel functional OA susceptibility variant was identified by targeted re-sequencing. This variant locates in a predicted regulatory site and creates a recognition sequence for ELF3 and HMGA1 transcription factors that are predicted to play a significant role in articular cartilage homeostasis.

Effects of diethylhexyl phthalate (DEHP) on INSL3 mRNA expression by Leydig cells derived from mouse embryos and in newborn mice.

Insulin-like factor 3 (INSL3) regulates testicular descent during fetal life, and Insl3 gene inactivation results in cryptorchidism. Little is known, however, about whether the plasticizer diethylhexyl phthalate (DEHP), a contaminant found widely in the environment, influences INSL3 expression. In this study, primary cultures of Leydig cells from mouse embryos were treated in vitro with DEHP. We also treated pregnant mice with DEHP from gestation day 12 to postnatal day 3 in order to study the effect of DEHP in vivo. INSL3 mRNA expression levels in primary Leydig cell cultures and in the testes of newborn mice were significantly lower following DEHP treatment. DEHP also caused detrimental morphological changes in both primary cultures of Leydig cells and the testes of newborn mice. These results suggest that the downregulation of INSL3 mRNA by DEHP might cause abnormalities of gubernacular development, which might be one of the mechanisms for development of cryptorchidism.

[Study on intergenera fusion of protoplasts from Rhizobium leguminosorum and Sinorhizobium xinjiangnesis].

Penicillin and chloromycetin were regarded as the sign of resistance to antibodies of R. leguminosorum USDA2370 and S. xinjiangnesis CCBAU110 respectively. Using the protoplast fusion technique, USDA2370 and CCBAU110 were successfully fused. Fusion hybrid can inoculate in the leguminous of parental strains respectively. There were apparent differences between parents and fusion hybrid in cell morphology, colony and pattern of whole-cell protein. The values of DNA homology between fusion hybrid and USDA2370 and CCBAU110 were 56.6% and 10.2% respectively.

[Analysis of 16S rDNA sequence and DNA-DNA hybridization of rhizobia isolated from Indigofera sp].

Based on the previous studies on numerical taxonomy and SDS-PAGE of whole-cell protein, the rhizobia strains isolated from Indigofera sp. in loess plateau area of North-west China constituted a new cluster, the 16S rDNA sequence of representative strain SHL042 was tested, and the phylogenetic tree was produced. In this tree, the strain SHL042, R. tropici A, R. tropici B, R. leguminosarum, R. etli, R. hananesis, R. mongolense and R. gallicum constituted a branch of phylogenetic. Within this branch, the similarity values of 16S rDNA sequences were 95.4%, 95.5%, 96.3%, 95.8%, 96.3%, 97.9% and 97.7% respectively. The values were more than 95%. This indicated that these species should belong to the same genus. The values of DNA homology in the new cluster were all more than 80%, but the values between SHL042 and the strains of these species were less than 50%. Thus, the strain SH714 represented a new rhizobia species.