Prevalence and prognostic significance of main metabolic risk factors in primary biliary cholangitis: a retrospective cohort study of 789 patients.

Background: Metabolic risk factors in primary biliary cholangitis (PBC) have not been well described in China. Additionally, it is unclear whether these factors have an impact on the prognosis of PBC patients. Therefore, this study aimed to investigate the prevalence of main metabolic risk factors in PBC, and to evaluate their prognostic values for liver-related outcomes. Methods: A cohort of 789 PBC patients was retrospectively studied between July 2008 and September 2019 by investigating the main metabolic risk factors and analyzing liver-related outcomes. Results: At presentation, 271 (34.3%) patients had concomitant hyperlipidemia, 126 (16.0%) had hypertension, 94 (11.9%) had type 2 diabetes mellitus (T2DM), and 17 (2.2%) had nonalcoholic fatty liver disease (NAFLD). Hyperlipidemia was found to be associated with the lower risk of liver-related death [P<0.0001, hazard ratio (HR): 0.397, 95% confidence interval (CI): 0.268-0.588] and adverse outcomes (P<0.0001, HR: 0.487, 95% CI:0.367-0.646), while hypertension was noted as a risk factor for liver-related death (P=0.001, HR: 1.788, 95% CI:1.268-2.521) and adverse outcomes (P=0.014, HR: 1.417, 95% CI:1.074-1.869). Moreover, age ≥ 55 years old (P=0.005) and cirrhosis (P<0.0001) had superimposition effects on hypertension as a risk factor for liver-related death, while only cirrhosis (P<0.0001) had an effect on hypertension as a risk factor for adverse outcomes. Additionally, anti-sp100 was associated with adverse outcomes (P=0.013) in PBC patients with hypertension in univariate Cox regression analysis. Conclusion: Hyperlipidemia, hypertension, and T2DM were found as main metabolic risk factors in PBC in China. Hyperlipidemia indicated a benign clinical outcome of PBC, while hypertension indicated a poor outcome of PBC. Older age and cirrhosis had superimposition effects on hypertension for liver-related poor outcomes. Anti-sp100 might be associated with adverse outcomes, especially in PBC patients with hypertension.

Disease predisposition of human leukocyte antigen class II genes influences the gut microbiota composition in patients with primary biliary cholangitis.

Background: The human leukocyte antigen (HLA) susceptibility gene is the main genetic risk factor for primary biliary cholangitis (PBC). The prognosis of patients with PBC is linked to gut microbiota dysbiosis. However, whether the HLA alleles are associated with the gut microbiota distribution and disease severity remains unknown. Methods: A cohort of 964 Chinese patients with PBC was enrolled at Beijing YouAn Hospital, Beijing, China. High-resolution genotyping of the HLA class I and class II loci from 151 of these patients was performed using sequence-based PCR. Stool samples were collected from 43 of the 151 fully HLA-typed patients to analyze their microbiota compositions via 16S RNA gene sequencing. Results: Of the 964 patients, the male:female ratio was 114:850, and 342 of these patients (35.5%) had already developed liver cirrhosis (LC) before enrollment. Patients with PBC showed a significantly higher frequency of HLA DRB1*08:03 than did the controls (21.2% vs. 9.0%, P=0.0001). HLA-DRB1*03:01, DRB1*07:01, DRB1*14:05, and DRB1*14:54 frequencies were also increased but did not reach significance after Bonferroni's correction. Conversely, the DQB1*03:01 frequency was significantly lower in patients with PBC than in the controls (24.5% vs. 39.2%, P=0.0010). The patients' gut microbiota were analyzed from four perspectives. The microbial community abundances were significantly lower in FHRAC-positive patients (patients with a combination of five HLA DRB1 high-risk alleles) than in FHRAC-negative patients (P<0.05). Of the top 10 microbial genera, Lachnospiraceae_incertae_sedis was higher in the FHRAC-positive patients than in the FHRAC-negative patients (P<0.05). linear discriminant analysis (LDA) effect-size (LEfSe) analysis showed different microbes at different levels in the FHRAC-negative patients but not in the FHRAC-positive patients. DQB1*03:01-positive patients contained mostly Lactobacillaceae at the family level. A comparison of the FHRAC-positive patients with and without liver cirrhosis showed that the abundances of Veillonella were significantly higher in patients with cirrhosis and FHRAC than in those without cirrhosis and are FHRAC-negative. Conclusion: The HLA class II genes may influence the gut microbiota compositions in patients with PBC. Differential gut microbiota were expressed at different taxonomic levels. Some bacterial abundances may be increased in FHRAC-positive patients with PBC and cirrhosis.

Using two-step cluster analysis to classify inpatients with primary biliary cholangitis based on autoantibodies: A real-world retrospective study of 537 patients in China.

Background: A variety of autoantibodies have been detected in primary biliary cholangitis (PBC), while the presence of autoantibody clusters and their clinical significance have not been fully understood. We aimed at defining autoantibody clusters and to better understand the clinical features and prognosis of PBC patients based on autoantibody clusters under real-world conditions. Methods: We retrospectively analyzed 788 inpatients with PBC evaluated between October 2008 and July 2019, and included 537 patients. Nineteen autoantibodies which were measured routinely were investigated for cluster analysis. Two-step clustering, Kaplan-Meier survival, and Cox regression analyses were used. Results: Five clusters were defined. A cluster of antinuclear antibodies (ANA) and anti-gp210 positive patients were identified with a high rate of cirrhosis at baseline and low survival rate; a cluster of ANA, anti-centromere antibodies (ACA) and/or anti-CENP-B female dominant patients with older disease onset, low level of platelet count at baseline, high rate of hepatic decompensation, and low survival rate was also characterized; and another cluster of anti-mitochondrial antibodies (AMA) and/or AMA-M2, anti-Ro52 and a high rate of anti-gp210 positive patients were identified with a high proportion of male patients and low survival rate. A subgroup of patients with anti-SSA and/or anti-SSB coexists with SjS was also identified; patients with only AMA and/or AMA-M2-positive with a benign clinical outcome and relatively high complication of non-alcoholic fatty liver disease (NAFLD) were also identified. Only anti-gp210 was considered as a significant predictor for poor outcomes especially in patients with cirrhosis. Conclusion: Clustering methods allow the identification of distinct autoantibody profiles of PBC that form clinical subsets and can be useful for personalized approaches to diagnosis, clinical management, and the prediction of clinical outcomes. Anti-gp210 was the strongest predictive factor for poor outcomes especially in PBC patients with cirrhosis under real-world conditions.

Development a multi-immunoaffinity column LC-MS-MS method for comprehensive investigation of mycotoxins contamination and co-occurrence in traditional Chinese medicinal materials.

Mycotoxins are poisonous secondary fungal toxic metabolites and harmful to human health. Traditional Chinese medicinal materials (TCMs), including more than two hundred functional foods, are vulnerably bred fungi, causing spoilage and multi-mycotoxins contamination. This study established a simultaneous analytical method by using multi-mycotoxins immunoaffinity column (multi-IAC) and HPLC-MS/MS to evaluate mycotoxins' contamination levels and natural incidence in TCMs. Aflatoxins (AFs, including AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), fumonisins (FB1 and FB2), zearalenone (ZEN), deoxynivalenol (DON) and T-2 toxins in three TCMs or functional foods of Polygalae Radix (PR), Coicis Semen (CS) and Eupolyphaga Steleophaga (ES) were detected. The systematically investigated results of 30 batch AFB1 positive samples revealed co-occurrence and correlation of multi-mycotoxins are significant differences in various matrices. All the samples in this study contain more than 5 mycotoxins. AFB1-AFs, AFB1-FBs, AFB1-DON, and AFB1-T-2 are the most observed co-occurrence, AFB1-OTA is also of concern due to its synergistic toxicity. This study's results can be used to establish guidelines for screening mycotoxin contaminants and limitations on acceptable levels in TCMs. Simultaneously, mycotoxin's correlation results in different matrices can also provide a reference for the standardization of TCM production and processing.

Clinical characteristics and HLA genotypes in Chinese patients with anti-SLA/LP-positive autoimmune hepatitis.

BACKGROUND: Anti-soluble liver antigen/liver pancreas (anti-SLA/LP) is a highly specific serological marker for the diagnosis of autoimmune hepatitis (AIH). The aim of the present study was to define the clinical characteristics and human leucocyte antigen (HLA) genotypes of Chinese patients with anti-SLA/LP positive AIH. METHODS: Ninety-one AIH patients who were anti-SLA/LP positive were enrolled in this case control study. Clinical information was obtained through reviewing patients' clinical notes. High-resolution genotyping of HLA-A, B, C, DRB1, and DQB1 alleles was performed by sequence-based typing polymerase chain reaction on 62 of the 91 patients. Data from 500 healthy patients were used as baseline controls. RESULTS: Anti-SLA/LP-positive AIH patients were characterized as follows: adults (age 20-80 years), female (88%), and frequent anti-nuclear antibody positivity (91%). Genetically, compared with the controls, HLA-B*35:01 and C*08:01 were significantly more frequent in patients. The frequencies of HLA-B*08:01, B*40:02, DRB1*04:01, DRB1*04:05, DRB1*14:01, and DRB1*16:02 increased, and the frequency in DRB1*15:01 decreased in patients, but did not reach significance after Bonferroni's correction. Patients with other autoimmune diseases had a higher DRB1*04:05 and DQB1*04:01 allele carrier frequency than those without. DRB1*04:05 and DQB1*04:01 alleles were found at increased frequency in patients with decompensated liver disease than those with compensated liver disease. CONCLUSIONS: Chinese anti-SLA/LP-positive AIH patients have some distinct clinical characteristics than other populations reported in the literature. The presence of certain specific HLA alleles could potentially increase the risk of developing anti-SLA/LP-positive AIH or other autoimmune disease and decompensated liver disease in the Chinese population.

Receptive and Expressive Vocabulary Skills and Their Correlates in Mandarin-Speaking Infants with Unrepaired Cleft Lip and/or Palate.

BACKGROUND: Vocabulary skills in infants with cleft lip and/or palate (CL/P) are related to various factors. They remain underexplored among Mandarin-speaking infants with CL/P. This study identified receptive and expressive vocabulary skills among Mandarin-speaking infants with unrepaired CL/P prior to cleft palate surgery and their associated factors. METHODS: This is a cross-sectional study involving patients at the Cleft Lip and Palate Center of the Stomatological Hospital of Xi'an Jiaotong University between July 2017 and December 2018. The Putonghua Communicative Development Inventories-Short Form (PCDI-SF) was used to assess early vocabulary skills. RESULTS: A total of 134 children aged 9-16 months prior to cleft palate surgery were included in the study. The prevalences of delays in receptive and expressive vocabulary skills were 72.39% (95% CI: 64.00-79.76%) and 85.07% (95% CI: 77.89-90.64%), respectively. Multiple logistic regression identified that children aged 11-13 months (OR = 6.46, 95% CI: 1.76-23.76) and 14-16 months (OR = 24.32, 95% CI: 3.86-153.05), and those with hard/soft cleft palate and soft cleft palate (HSCP/SCP) (OR = 5.63, 95% CI: 1.02-31.01) were more likely to be delayed in receptive vocabulary skills. CONCLUSIONS: Delays in vocabulary skills were common among Mandarin-speaking CL/P infants, and age was positively associated with impaired and lagging vocabulary skills. The findings suggest the necessity and importance of early and effective identification of CL/P, and early intervention programs and effective treatment are recommended for Chinese CL/P infants.

Profiling of the pattern of the human TRB/IGH-CDR3 repertoire in primary biliary cholangitis patients.

INTRODUCTION: Primary biliary cholangitis (PBC) is characterized by lymphocyte cell-induced immune destruction of cholangiole. However, the immunological characteristics of peripheral blood cells in PBC patients remain unknown. This study was designed to reveal the differences in the immunological characteristics between PBC patients and healthy adults. METHODS: We performed high-throughput sequencing to determine the TRB-CDR3 and IGH-CDR3 repertoires of T and B cells in 19 healthy controls and 29 PBC patients. Different immunological characteristics, such as distinctive complementarity determining region 3 (TRB-CDR3) lengths, usage bias of V and J segments, and random nucleotide addition were identified in PBC and healthy control (HC) groups. RESULTS: The diversity of TRB-CDR3 was significantly lower in the PBC group compared with the HC group. CDR3 and the N addition length distribution were significantly changed compared with the HC group. It appeared that the PBC group had more short N additions and the HC group had more long N additions in the TRB-CDR3 repertoire. The results also revealed a set of PBC-associated clonotypes compared with the HC group. CONCLUSION: This study suggested that PBC is a complex autoimmune disease process with evidence of different TRB-CDR3 rearrangements compared with healthy adults that share IGH-CDR3 peptides with some autoimmune diseases. This new insight may contribute to a better understanding of the immune functions of PBC patients and benefit efficient applications of PBC diagnosis and treatments.

Fluctuations of antimitochondrial antibodies and anti-gp210 antibody in a patient with primary biliary cholangitis and Sjögren syndrome with subsequent autoimmune hemolytic anemia: A case report.

RATIONALE: Primary biliary cholangitis (PBC) is a rare autoimmune cholestatic liver disease. It is often associated with extrahepatic autoimmune disorders. However, the concurrence of PBC and Sjögren syndrome (SS) with the subsequent onset of autoimmune hemolytic anemia (AIHA) is extremely rare. PATIENT CONCERNS: This study investigated a 60-year-old woman admitted to our hospital with complaints of xerostomia for 5 years, pruritus for 3 years, and abnormal liver function for 3 months. DIAGNOSES: The patient was suffering from typical clinical PBC and SS, and developed decompensated liver cirrhosis after 32 months of ursodeoxycholic acid (UDCA) therapy. In May 2018, she was readmitted to the hospital with a high fever of 39 °C, coughing, and sever fatigue without remission after 3 days of cephalosporin antibiotic therapy. During the clinical course of PBC, her antimitochondrial antibodies (AMA) titers fluctuated from 1:1000 to negative and then to weakly positive, determined by indirect immunofluorescence (IIF), immunoblotting, and enzyme-linked immunosorbent assay (ELISA) based on recombinant mitochondrial antigens; furthermore, her titers of anti-gp210, an antinuclear antibody (ANA), increased sharply. Laboratory tests and imaging were performed to diagnose PBC and SS in September 2015. However, she was subsequently diagnosed with AIHA after 32 months of UDCA therapy based on the identification of pancytopenia, increased reticulocyte (RET) count, and a positive result from the direct Coombs test. INTERVENTIONS: UDCA, hepatic protectant, albumin infusion, chest drainage, rational antibiotic use, diuretics, and methylprednisolone were used to treat the patient. OUTCOMES: Liver cirrhosis was complicated by the development of AIHA, which became severe at 42 months of follow-up. LESSONS: This is the first case report showing a patient with comorbid PBC and SS, as well as the sequential development of AIHA with decreased AMA and increased anti-gp210 titers; this may have been due to immunodeficiency. These findings stress the importance of the serological screening of ANA profile, as well as repeated measurement of ANA and AMA to track PBC progression and prognosis.

[Effect of Chinese drugs for Pi strengthening Shen benefiting on the immunity function of HIV patients' specific T cells].

OBJECTIVE: To explore the effect of Chinese drugs for Pi strengthening Shen benefiting (CDPSSB) on the immunity function of HIV/AIDS patients' specific T cells. METHODS: Totally 20 patients were randomly recruited from the treated group [treated by CDPSSB combined highly active anti-retroviral therapy (HAART)] and 23 patients were randomly recruited from the control group (treated by HAART alone). All patients were follow-up infected persons form You'an Hospital between from June 2010 to June 2012. CD4+ T absolute counts and HIV viral load were detected. Meanwhile, HIV whole gene overlapping peptides were used as stimulating antigen. The response intensity of HIV specific T cells was detected in the two groups. RESULTS: There was no statistical difference in CD4 T absolute counts or HIV viral load between the two groups (P > 0.05). The response intensity of HIV specific T cells was significantly enhanced in the treated group, when compared with the control group (P < 0.05). Along with elongation of treatment time (6, 12, 18, and 24 months) in the treated group, the response intensity of HIV specific T cells showed enhancing tendency, but there was no statistical difference among these time points (P > 0.05). CONCLUSION: CDPSSB could enhance improve the immunity function of HIV specific T cells, which might be one of its mechanisms.

Human leucocyte antigen alleles and haplotypes and their associations with antinuclear antibodies features in Chinese patients with primary biliary cirrhosis.

BACKGROUND & AIMS: Primary biliary cirrhosis (PBC) is an autoimmune liver disease. Genetic factors are critical in determining susceptibility to PBC. Among human leuocyte antigen (HLA) genes, an association between the DRB1*08 allele and PBC has been reported in many populations, but not in Chinese patients. METHODS: We investigated HLA-A, B, DRB1, and DQB1 alleles and haplotypes in 145 PBC patients and 500 healthy subjects. Patients were also stratified according to autoantibody features, and associations between these and HLA alleles were analyzed. RESULTS: Significant associations existed between HLA-DRB1*08:03 (22.1% vs. 9.0%, Pc < 0.0001, OR = 2.86), DQ2 (41.4% vs. 25.4%, Pc < 0.0001, OR = 2.07) and DQB1*06:01 (31.0% vs. 17.8%, Pc = 0.014, OR = 2.08) alleles and PBC. DRB1*08:03-DQB1*06:01 (22.1% vs. 8.2%, P < 0.0001, OR = 3.17) and DRB1*07:01-DQB1*02:02 haplotypes (28.3% vs. 17.6%, P = 0.005, OR = 1.85) were also associated with PBC susceptibility. In contrast, the DQB1*03:01 allele (21.4% vs. 39.2%, Pc < 0.0001, OR = 0.42) and DRB1*12:02-DQB1*03:01 haplotype (6.9% vs. 14.6%, P = 0.015, OR = 0.43) were significantly decreased in PBC patients compared with controls. DRB1*14:54 and DQ5(1) protected against antinuclear antibody (ANA) (OR = 0.25) and anti-gp210 antibody (OR = 0.39) production, respectively, while HLA-B*44:03 predisposed patients to anti-gp210 antibody (OR = 5.70) production. CONCLUSION: These results suggest that Chinese patients with PBC have a distinct genetic background in eastern Asia, and we confirmed the role of HLA genes in determining PBC susceptibility and autoantibody features in the Chinese population.

[Investigate circulating levels of chemokines and evaluate the correlation between these chemokines and liver function indicators in autoimmune hepatitis].

OBJECTIVE: This study investigated circulation levels of chemokines (CCL2, CCL5, CXCL8, CXCL9, CXCL10) in autoimmune hepatitis(AIH) patients and evaluated the correlation between these chemokines and liver function indicators. METHODS: A total of 5 chemokines (CCL2, CCL5, CXCL8, CXCL9, CXCL10) were measured simultaneously by cytokine beads assay(CBA) in the sera of 46 patients with AIH and 12 cases of healthy control. RESULTS: In this study we found that serum levels of CCL2 , CXCL9 and CXCL10 in AIH patients and healthy controls were 11.79:8.39 pg/ml, 11.31:2.69 pg/ml, 15.85:4.64 pg/ml, respectively , which implied these chemokines were significantly higher in AIH patients when compared to healthy control (Z=-1.958, P=0.05; Z=-4.527, P less than 0.0001; Z=-3.84, P less than 0.0001, respectively). And circulation levels of CCL2 , CXCL8 , CXCL9 and CXCL10 in pretreatment and remission stages of patients with AIH were 29.69:11.16 pg/ml, 7.2:5.38 pg/ml, 16.02:5.47 pg/ml, 90.01:13.24 pg/ml, respectively, which showed these chemokines decreased during remission from pretreatment stage levels (t=2.985, P=0.005; Z=-2.547, P=0.0112; Z=-3.187, P=0.001; t=2.12, P=0.0015, respectively). Among AIH , CXCL8 was correlated positively with lgG(r2=0.291, P=0.0039); CXCL9 was associated positively with ALT and AST(r2=0.5324 , P less than 0.0001; r2=0.3352, P less than 0.0001); CXCL10 showed a positive correlation with ALT , AST and GGT(r2=0.9551, P less than 0.0001; r2=0.8960, P less than 0.0001; r2=0.8271, P less than 0.0001). CONCLUSION: Serum levels of CCL2, CXCL8, CXCL9 and CXCL10 are significantly higher in patients with AIH, but decrease to levels in healthy controls after successful treatment , and circulation levels of CXCL9 and CXCL10 are associated positively with liver function indicators which can react inflammation activity of liver, all these may imply that chemokines can reflect the degree of liver inflammation and may be one of the main culprits in AIH pathological damage.

[Prognostic values of chemokines in different clinical outcomes of influenza A (H1N1)-infected patients].

OBJECTIVE: To analyze the prognostic values of chemokines in different clinical outcomes of influenza A (H1N1)-infected patients. METHODS: A total of 60 cases with influenza A (H1N1) virus were enrolled. There were 32 mild cases, 16 healing cases from severe stage and 12 mortality cases resulting from severe stage. The serum levels of chemokines including CXCL8, CCL10, CCL2, CCL3, CCL4, CCL5 and CCL11 were detected by the Luminex technique. RESULTS: The levels of CXCL8, CCL2 and CCL10 in the mortality group were higher than those of the healing serve and mild cases. And the differences were significant. P value was 0.001, < 0.0001 and 0.027 respectively. The thresholds of CCL2 and CXCL8 for predicting clinical mortality were 45.99 ng/L (sensitivity 100%, specificity 89.58%) and 59.75 ng/L (sensitivity 75%, specificity 95.83%). CONCLUSION: The serum levels of chemokines are elevated in severe cases of influenza A (H1N1). And the rises of CXCL8 and CCL2 are more obvious in the mortality cases. Thus CXCL8 and CCL2 may serve as two prognostic indices for the clinical fatal outcomes.

[The analysis of effect of Th1/Th2 cytokine in the different prognosis in severe influenza A (H1N1)].

OBJECTIVE: To explore the effect of cytokine in the different prognosis of patients with severe influenza A (H1N1) infection. METHODS: 28 cases with severe influenza A (H1N1) were enrolled in the study including 16 cured cases and 12 dead cases. The cytokine level in serum was detected by Luminex technology. RESULTS: The levels of IL-2, IL-12 (P70) and IFN-gamma in dead group was lower than cured and normal control group and the difference were significant, P <0.05, respectively. IL-4 level in the dead group was significantly lower than cured group and normal control group, P value was 0.0310 and 0.0012, respectively. CONCLUSIONS: The Thl cytokine level in the severe 2009 epidemic H1N1 influeaze cases shows decreased trend, and the trend is more obvious in dead cases. The decrease of Th1 cytokine may be one of reasons leading to severe clinical situation and related withthe bad prognosis.

Prognostic factors and survival analysis of antimitochondrial antibody-positive primary biliary cirrhosis in Chinese patients.

BACKGROUND AND AIMS: Primary biliary cirrhosis (PBC) is a relatively uncommon liver disease, and information on the prognosis and survival of PBC patients in mainland China is lacking. We therefore conducted a retrospective study to investigate the prognostic factors and survival in Chinese PBC patients. METHODS: Between October 2001 and May 2009, patients registered at Beijing You'an Hospital with abnormal liver function and serum positivity for antimitochondrial antibody (AMA) and/or AMA-M2 (n = 391) were screened. Patients diagnosed with PBC were identified, and their medical data were reviewed and analyzed for mortality predictors. RESULTS: A total of 147 PBC patients were identified (mean age: 54 years, range: 28-81), of whom 126 (85.7%) were women. At the time of diagnosis, 119 patients (81.0%) were symptomatic, 28(19.0%) had hepatic decompensation, and no patients were asymptomatic. During a median follow-up period of 48 months (range: 2-312), 36 patients (24.5%) died or underwent liver transplantation, and 65 patients (44.2%) developed hepatic decompensation. The overall 5-year survival rate was 79%. Multivariate analysis indicated that Mayo risk score ≥6.11(P = 0.008), and serum IgG ≥ 17.20 g/l (P = 0.016) were associated with mortality. CONCLUSIONS: Most Chinese PBC patients in this study were symptomatic at diagnosis and had significant mortality. Mayo risk score, and serum IgG were independent prognostic factors for survival.

[Phylogenetic relationships among some Chinese genera of Phoridae based on mitochondrial 12S and 16S rRNA gene sequences].

The phylogenetic relationships among 14 Chinese genera of Phoridae were reconstructed based on concatenated sequences of the mitochondrial 12S and 16S rRNA genes using maximum parsimony, maximum likelihood and Bayesian methods. The alignment of the concatenated sequences spanned 819 sites including 277 variable sites, of which 200 were parsimony-informative. The sequences showed a 77.7% A+T bias. The phylogenetic analysis showed that the Phoridae was monophyletic. It was shown that the involved genera in the study clustered into monophyletic Phorinae and Metopininae. The genera Diplonevra and Dohrniphora were closely related to each other. The relationships among Anevrina, Conicera and Spiniphora were also closer. The genera Metopina and Puliciphora proved to be reciprocal sister group and the cluster analysis showed the close relationship between Gymnophora and Phalacrotophora.