Mendelian-randomization study revealed causal relationship between nonalcoholic fatty liver disease and osteoporosis/fractures.

BACKGROUND: Patients with nonalcoholic fatty liver disease (NAFLD) are reported to have a higher risk of osteoporosis/fractures; however, the causal relationship remains unclear. METHODS: Publicly available genome-wide association studies (GWASs) were used for Mendelian randomization (MR) analysis. GWASs of NAFLD and fractures were obtained from the FinnGen Consortium. GWASs of bone mineral density (BMD) were derived from a meta-analysis. GWASs of obesity, diabetes, liver function, and serum lipid-related metrics were used to clarify whether the accompanying NAFLD symptoms contributed to fractures. Moreover, two additional GWASs of NAFLD were applied. RESULTS: A causal association was not observed between NAFLD and BMD using GWASs from the FinnGen Consortium. However, a causal relationship between NAFLD and femoral neck-BMD (FN-BMD), a suggestive relationship between fibrosis and FN-BMD, and between NAFLD and osteoporosis were identified in replication GWASs. Genetically proxied body mass index (BMI), high-density lipoprotein (HDL), and hip circumference increased the likelihood of lower limb fractures. The waist-to-hip ratio decreased, whereas glycated hemoglobin (HbA1C) and homeostasis model assessment of ß-cell function (HOMA-B) increased the risk of forearm fractures. Low-density lipoprotein (LDL) reduced, whereas HbA1C increased the incidence of femoral fractures. Alkaline phosphatase (ALP) raised the risk of foot fractures. However, after a multivariate MR analysis (adjusted for BMI), all the relationships became insignificant. CONCLUSIONS: NAFLD caused reduced BMD, and genetically predicted HDL, LDL, HbA1C, HOMA-B, ALP, hip circumference, and waist-to-hip ratio causally increased the risk of fractures. BMI may mediate causal relationships. Larger GWASs are required to verify this finding.

Emergomyces orientalis Emergomycosis Diagnosed by Metagenomic Next-Generation Sequencing.

Emergomyces is a newly described dimorphic fungus genus; it may cause fatal infections in immunocompromised patients, but diagnosis is often delayed. We report a case of disseminated emergomycosis caused by the novel species Emergomyces orientalis in a kidney transplant recipient from Tibet. Infection was diagnosed early by metagenomic next-generation sequencing.

Safety and Efficacy of Regional Citrate Anticoagulation during Plasma Adsorption Plus Plasma Exchange Therapy for Patients with Acute-on-Chronic Liver Failure: A Pilot Study.

BACKGROUND: Patients with acute-on-chronic liver failure (ACLF) might be at risk for citrate accumulation during plasma adsorption plus plasma exchange (PE) therapy with regional citrate anticoagulation (RCA). OBJECTIVES: To assess the safety and efficacy of RCA during double plasma molecular adsorption system (DPMAS) plus PE therapy for patients with ACLF. METHOD: A prospective nonrandomized controlled pilot study was conducted at West China Hospital of Sichuan University. Patients with ACLF were enrolled to heparin anticoagulation (HA) group and RCA group. Serial blood samples were taken. Patients were followed up for 3 months. RESULTS: Twenty-four patients with 94 sessions of HA and 28 patients with 106 sessions of RCA were enrolled. RCA method did not affect the therapeutic efficacy, the function of extracorporeal circulation, and the prognosis of these patients. The occurrences of citrate accumulation in RCA group were 0.0, 67.0, 100.0, 34.0, and 0.0% before DPMAS therapy, at the end of DPMAS therapy, immediately after PE therapy, 2 h after PE therapy, and the next morning, while that in HA group were 0.0, 0.0, 100.0, 7.4, and 0.0%, respectively. The occurrences of citrate accumulation at the end of DPMAS therapy and at 2 h after PE therapy in RCA group were much higher than that in HA group (67.0 vs. 0.0%, p = 0.000; 34.0 vs. 7.4%, p = 0.000, respectively). Although the trend of citrate accumulation in RCA group was much more obvious than that in HA group during and after DPMAS plus PE therapy (p = 0.000), the values on the next morning were similar between the 2 groups (p > 0.05). The main alteration of acid-base status was metabolic alkalosis with no difference between the 2 groups. CONCLUSIONS: RCA might be safe and effective in patients with ACLF receiving plasma adsorption plus PE therapy. RCA method might offer an alternative anticoagulation method for them.

[The Application of Estrogen Receptor Aptamer in Immunohistochemical Detection of Breast Cancer].

OBJECTIVE: To synthesize and select an estrogen receptors aptamer that can be used in immunostaining of breast cancer tissues. METHODS: ER protein was purified. ER aptamer that showed a high affinity and specificity for ER was synthesized and selected and by SELEX. Ligand -receptor interactions assay was adopted to measure the affinity of the aptamer-ER complex. Both the biotinylated aptamer and the anti-ER monoclonal antibody were tested for immunohistochemical staining of ER status on 105 breast cancer samples. Agreement on the detection of ER expression was determined by Kappa statistics. RESULTS: The dissociation contant (Kd) of the biotinylated aptamer-ER complex, as calculated by a linear regression analysis, was determined to be (0.34±0.05) nmol/L ( n=3, r=0.989). The binding capacity (B max) was 769.23 fmol/(mg prot·nmol -1·L -1). The ER aptamer and the anti-ER antibody both exhibited identical specificity to ER-expressing breast cancer cells. There was a high agreement between the two methods ( n=105, Kappa value=0.943, 95% confident interval=0.879-1.006, P<0.05 for the ER positive and negtive samples; n=75, Kappa value=0.805, 95% confident interval=0.642-0.967, P<0.05 for the ER weak and moderate/strong expression samples). Both the anti-ER antibody and the ER aptamer can also recognized breast cancer cells at the same sites. There was no expression in the negative controls. There were also positive expressions in the 2 endometrial cancer tissues by using biotinylated aptamer. CONCLUSIONS: Our results indicated that the synthesized ER aptamer has a high affinity to bind ER. ER aptamer and the anti-ER antibody can both be used for immunohistochemical staining of ER status in breast cancer tissue.

Pronounced decline of serum HBsAg in chronic hepatitis B patients with long-term effective nucleos(t)ide analogs therapy.

AIMS: This study aims to investigate the kinetics of serum HBsAg levels in chronic hepatitis B patients with long-term nucleos(t)ide analogs (NAs) therapy. METHODS: This was a retrospective clinical study. Serum HBsAg in serial samples of 94 patients, who received at least 8 years of NAs therapy, were measured using Elecsys® HBsAg II Quant Assay. RESULTS: In this cohort, serum HBsAg levels reduced from 3.80 log10 IU/mL at baseline to 2.72 log10 IU/mL at year 8 (p < .001), and the percentage of patients with HBsAg <1000 IU/mL increased from 14.9% at baseline to 55.3% at year 8 (p < .001). The reduction of serum HBsAg did not differ significantly between patients stratified by baseline virological parameters and type of antiviral agents. But as compared to patients without HBeAg seroconversion, HBsAg levels were significant lower in patients with HBeAg seroconversion (3.19 vs. 2.47 log10 IU/mL at year 8, p = .001). As compared to patients with slow (0-1 log10 IU/mL) or steady HBsAg(≤0 log10 IU/mL) decline at year 1, patients with a rapid HBsAg (≥1 log10 IU/mL) decline had a significantly lower HBsAg levels from year 2 to 8. However, Cox regression analysis showed that only absolute HBsAg levels at year 1 was an independent predictor of subsequent HBsAg <1000 IU/mL at year 8 of antiviral therapy(HR 0.242, p = .004). CONCLUSION: Pronounced HBsAg declines could be achieved in patients after long-term effective therapy with NAs, and on-treatment low serum HBsAg level at year 1 might be a predictor of serum HBsAg <1000 IU/mL at year 8.

Comparison of clinical features and prognostic factors in HIV-negative adults with cryptococcal meningitis and tuberculous meningitis: a retrospective study.

BACKGROUND: The incidence of cryptococcal meningitis (CM) and tuberculous meningitis (TBM) have gradually increased in recent years. These two types of meningitis are easily misdiagnosed which leads to a poor prognosis. In this study we compared differences of clinical features and prognostic factors in non-HIV adults with CM and TBM. METHODS: We retrospectively reviewed the medical records of CM and TBM patients from January 2008 to December 2015 in our university hospital in China. The data included demographic characteristics, laboratory results, imaging findings, clinical outcomes. RESULTS: A total of 126 CM and 105 TBM patients were included. CM patients were more likely to present with headache, abnormal vision and hearing, and they might be less prone to fever and cough than TBM patients (P < 0.05). Higher percentage of CM patients presented with cerebral ischemia/infarction and demyelination in brain MRI than TBM patients (P < 0.05). CM patients had lower counts of WBC in CSF, lower total protein in CSF and serum CD4/CD8 ratio than TBM patients (P < 0.05). After three months of treatment, CM group have worse outcome than TBM group (P < 0.05). Multivariate analysis showed that age more than 60y (OR = 4.981, 95% CI: 1.955-12.692, P = 0.001), altered mentation (OR = 5.054, 95% CI: 1.592-16.046, P = 0.006), CD4/CD8 ratios < 1 (OR = 8.782, 95% CI: 2.436-31.661, P = 0.001) and CSF CrAg ≥ 1:1024 (OR = 4.853, 95% CI: 1.377-17.098, P = 0.014) were independent risk factors for poor prognosis for CM patients. For TBM patients, hydrocephalus (OR = 7.290, 95% CI: 1.630-32.606, P = 0.009) and no less than three underlying diseases (OR = 6.899, 95% CI: 1.766-26.949, P = 0.005) were independent risk factors, headache was a protective factor of prognosis. CONCLUSIONS: Our study provided some helpful clues in the differential diagnosis of non-HIV patients with CM or TBM and identified some risk factors for the poor prognosis of these two meningitis which could help to improve the treatment outcome. Further studies are worth to be done.

Meta analysis of the association of cholesterol with pancreatic carcinoma risk.

PURPOSE: Pancreatic carcinoma is a malignant tumor with poor prognosis. This metaanalysis was conducted to investigate if there exists any association of cholesterol with pancreatic carcinoma risk. METHODS: A literature search was performed in Cochrane Central Library, PubMed, MEDLINE, EMBASE, CNKI (China National Knowledge Infrastructure), China Biology Medical literature database (CBM), and WangFang database for relevant available articles. Dietary cholesterol and serum levels of total cholesterol (TC) were assessed and compared. Pooled relative risks (RRs) with 95% confidence intervals (CIs) were calculated. RESULTS: A total 19 articles coming from Europe, Asia and north America were assessed in this study. There was a significant difference between highest and lowest dietary cholesterol intake for pancreatic carcinoma risk (RR=1.31, 95% CI:1.10 to 1.56, p=0.01). Moreover, in subgroup analysis, there was a significant difference between highest and lowest dietary cholesterol intake for pancreatic carcinoma risk for case-control studies (RR=1.52, 95% CI:1.23 to 1.90, p=0.04). However, no significant difference was noticed between highest and lowest dietary cholesterol intake for pancreatic carcinoma risk for cohort studies (RR=1.02, 95% CI:0.87 to 1.20, p=0.51). The meta analysis results showed a significant difference between highest and lowest dietary cholesterol for pancreatic carcinoma in Europeans (RR=1.15, 95% CI:0.86 to 1.53, p=0.05). Moreover, compared to the low serum level of TC, the high level serum TC was associated with pancreatic carcinoma risk (RR=1.00, 95% CI:0.86-1.17, p=0.03). There was a significant difference between high and low levels of serum TC for pancreatic carcinoma risk in Europeans (RR=1.03, 95% CI: 0.72 to 1.48, p=0.04). CONCLUSION: Dietary or serum cholesterol may be associated with risk for increased pancreatic carcinoma.

[Association of chronic obstructive pulmonary disease and lung cancer].

OBJECTIVE: To investigate the association between chronic obstructive pulmonary disease (COPD) and lung cancer. METHODS: A case-control study was undertaken, with 180 cases of lung cancer and 200 cases of controls. RESULTS: The odd of lung cancer was higher in patients with COPD, emphysema, chronic bronchitis, and pulmonary tuberculosis (P < 0.05). The odd of lung cancer increased significantly in patients with a family history of lung cancer or COPD (P < 0.05). The odd of lung cancer also increased when forced expiratory volume in one second (FEV1) < 80%. CONCLUSION: Patients with COPD or a family history of COPD have higher risk of lung cancer.

Progressive multifocal leukoencephalopathy in an HIV patient: A case report and literature review.

Key Clinical Message: Progressive multifocal leukoencephalopathy (PML) is a rare opportunistic infection of the brain caused by reactivation of the JC virus, which can lead to a lytic infection of oligodendrocytes. We report a patient with HIV who developed PML. Abstract: Progressive multifocal leukoencephalopathy (PML) is a rare opportunistic infection of the brain caused by reactivation of the John Cunningham virus (JCV), which can lead to a lytic infection of oligodendrocytes. Herein, we report the case of a patient with HIV who developed PML that presented as a progressive disturbance of consciousness and movement. The patient's clinical symptoms progressively deteriorated, and positive JC viral DNA in his cerebrospinal fluid (CSF) helped us diagnose him with PML. Magnetic resonance imaging (MRI) showed multiple asymmetric subcortical and deep white-matter lesions. Although we administered immunoreconstructive therapy, the patient's condition gradually worsened. Therefore, we suggest that PML should be considered if such lesions are found in MRIs of HIV patients.

Construction of a plasmid vector for liver-specific inhibition of hepatocyte nuclear factor 4 alpha expression.

Hepatocyte nuclear factor-4alpha (HNF-4a) is an important transcription factor in the liver, and regulates a large number of genes involved in many aspects of hepatocyte functions. In this study, a liver-specific transcriptional regulatory element comprised of albumin promoter (ALBp) and alpha-fetoprotein enhancer (AFPe) was obtained and cloned into the plasmid pHNF4sh-CMV(short hairpin RNA targeting HNF4α) with original CMV promoter removed, resulting to pHNF4sh-EP for liver-specific knockdown of HNF4α expression. In an attempt to verify its characteristics, pHNF4sh-EP was transfected to L02, HepG2, and COS1 cell lines in vitro and delivered into mice in vivo. pHNF4sh-CMV and pNCsh-EP were used as controls. For in vitro, the level of HNF4α mRNA and protein was decreased in all cell lines transfected with pHNF4sh-CMV whereas HNF4α mRNA and protein decreasing was only observed in L02 and HepG2 cell lines upon transfection with pHNF4sh-EP, and this decreasing was more significant as compared with pHNF4sh-CMV transfected cells. For in vivo, the decreasing of HNF4α mRNA and protein was observed in both liver and kidney tissues upon transfection with pHNF4sh-CMV. After transfection with pHNF4sh-EP, decreasing of HNF4α mRNA and protein was only found in liver tissue and this decreasing was more significant. No obvious HNF4α mRNA and protein decreasing was detected either in vitro or in vivo after transfected with pNCsh-EP. In conclusion, pHNF4sh-EP could highly-active and liver-specific knockdown of HNF4α expression liver and it will be useful for further study of the funcitions of HNF4α in liver.

A mutation in the interferon regulatory element of HBV may influence the response of interferon treatment in chronic hepatitis B patients.

BACKGROUND: A functional interferon regulatory element (IRE) has been found in the EnhI/X promoter region of hepatitis B virus (HBV) genome. The purpose of this study is to compare the gene order of responder and non-responder to interferon therapy in patients with chronic hepatitis B (CHB), so as to evaluate the relationship between IRE mutation and the response to interferon treatment for CHB patients. RESULTS: Synthetic therapeutic effect is divided into complete response (CR), partial response (PR) and non-response (NR). Among the 62 cases included in this study, 40 cases (64.5%) were in the response group (CR and PR) and 22 (35.5%) cases were in the NR group. Wild type sequence of HBV IRE TTTCACTTTC were found in 35 cases (56.5%), and five different IRE gene sequences. included TTTtACTTTC, TTTCAtTTTC, TTTtAtTTTC, TTTtACTTTt and cTTtACcTTC, were found in 22 cases (35.5%), 1 case (1.6%), 1 case (1.6%), 2 cases (3.2%) and 1 case (1.6%) respectively. There were 41.9%cases (26/62) with forth base C→T mutation, consisted of 32.5% (13/40) cases in response group and 59.1% (13/22) cases in NR group. Among the 35 cases with IRE sequences, there were 67.5% (27/40) cases in response group and 36.4% (8/22) in NR group, and the difference in IRE sequences between two groups was statistic significantly (P = 0.027). The result suggested that there is likely relationship between the forth base mutation (C→T) of IRE region and the response of HBV to Interferon therapy, and this mutation may partially decrease the inhibition effect of interferon on HBV. CONCLUSION: The forth base C→T mutation in IRE element of HBV may partially influence the response of Interferon treatment in CHB patients.

Inhibition of hepatitis B Virus replication by hepatocyte nuclear factor 4-alpha specific short hairpin RNA.

BACKGROUND: Previous studies showed that hepatocyte nuclear factor 4α (HNF4α) may play a critical role in hepatitis B virus (HBV) replication. AIMS: This study aimed to investigate the effect of knocking down of HNF4α with RNA interference technique on HBV replication in a HBV replication mouse model. METHODS: Four HNF4α, specific short hairpin RNA (shRNA)-producing plasmids were constructed. HBV mRNA and DNA replication intermediates were analysed using Northern and Southern blot respectively. The expression of HNF4α and HBV core antigen (HBcAg) was detected using immunohistochemistry technique. RESULTS: One of the HNF4α shRNAs, HNF4α shRNA1, efficiently inhibited the expression of HNF4α in HepG2 cells and mice liver. HBV RNA transcripts and DNA replication intermediates in HNF4α shRNA1 group were decreased 67.3 and 76%, respectively, in HepG2 cells, and 68.1 and 70.6% in mice liver respectively. The expression level of HBcAg in the liver was also decreased with the inhibition of HNF4α expression. CONCLUSIONS: These results suggested that decreasing of HNF4α expression was associated with the reduced level of HBV replication in HepG2 cells and mice liver. These data indicated that HNF4α played a critical role in HBV replication in vivo, and HNF4α shRNA could inhibit HBV replication in vivo.

Pre-core/basal-core promoter and reverse transcriptase mutations in chronic HBV infected-patients.

BACKGROUND/AIMS: Some HBV mutations have been shown to have an association with liver disease. The aim of the study was to investigate the incidence of mutations in hepatitis B virus (HBV) pre-core/basal core promoter (BCP) and reverse transcriptase (RT) regions and their relationship with disease progression in chronic HBV-infected patients. METHODOLOGY: A total of 133 patients were enrolled in this study, comprising the acute-on-chronic hepatitis B liver failure (ACLF-HBV) and chronic hepatitis B (CHB) patients. The pre-core/ BCP and RT gene fragments were amplified by high-fidelity PCR. Mutations of pre-core/BCP and RT regions were examined by direct sequencing. RESULTS: There were no significant differences in age, the average level of ALT and course of disease between the ACLF-HBV and CHB groups. The HBeAg positive rate and average values of HBV-DNA loads of the ACLF-HBV patients were lower than that of CHB patients. In HBV pre-core/ BCP region, the point mutations T1753C (39.06% vs. 21.74%, p<0.01), A1762T (26.56% vs. 13.04%, p<0.05), G1764A (31.25% vs. 18.84%, p<0.01), G1896A (29.69% vs. 15.94%, p<0.05) and G1899 (23.44% vs. 10.14%, p<0.05) were significantly more frequent in the ACLFHBV than CHB patients. For combined mutations, A1762T+G1764A (23.43% vs. 11.59 %, p<0.05) and G1896A+ G1899A (21.88% vs. 13.04%, p<0.05) were significantly more frequent in ACLF-HBV than CHB patients. However, there were no significant differences in RT mutations between two groups. CONCLUSIONS: ACLFHBV patients had more frequent mutations in HBV precore/ BCP region than that of CHB patients. Some mutations in HBV pre-core/BCP region might be related to the aggravation of chronic HBV infection.

Dynamics of Serum Pregenome RNA in Chronic Hepatitis B Patients Receiving 96-Month Nucleos(t)ide Analog Therapy.

Background: Tissue covalently closed circular DNA (cccDNA) can reflect the activity of HBV replication. However, it is impractical to assess intrahepatic cccDNA in every outpatient. Serum pregenome RNA (pgRNA) is transcribed from intrahepatic cccDNA and may reflect the activity of intrahepatic cccDNA. We explored the dynamics and the potential role of serum pgRNA in patients receiving long-term NAs treatment. Methods: Serum pgRNA, HBV DNA, HBsAg, HBeAg, and ALT levels were quantified, and the relationships between serum pgRNA and these common clinical indicators before and after the treatment were investigated. Results: Serum pgRNA showed dynamic change during the 96-month NAs therapy, and serum pgRNA levels were positive and detectable in 19 patients with undetectable serum HBV DNA. Serum pgRNA showed strong and positive correlation with serum HBV DNA (r = 0.693, p < 0.001) and serum HBsAg levels (r = 0.621, p < 0.001) at baseline. Patients with HBeAg seroconversion had lower baseline serum pgRNA levels (p = 0.002). The area under the curve (AUC) of baseline serum pgRNA for predicting HBeAg seroconversion was 0.742 (95% CI: 0.606-0.850) with 63.16% sensitivity and 80.56% specificity. The cumulative HBeAg seroconversion rate was higher in patients with low serum pgRNA (p = 0.001). Conclusion: Serum pgRNA of low level at baseline or great decline at month 6 may independently predict the high incidence of undetectable serum pgRNA at year 4 following NAs therapy, and the baseline serum pgRNA may serve as a novel predictor for HBeAg seroconversion during NAs therapy.

Construction of a highly-active, liver-specific transcriptional regulatory element through combination of the albumin promoter and α-fetoprotein enhancer.

In an attempt to construct a highly active, liver-specific transcriptional regulatory element, the mouse albumin promoter (ALBp) and α-fetoprotein enhancer (AFPe) were obtained. To verify its hepatic specificity and activity, the AFPe-ALBp-containing fragment was cloned into the plasmids, pVAX-S and pGL3-Luc with original promoter removed. Plasmid pVAX-AFPe-ALBp-S was then transfected into hepatic and non-hepatic cells in vitro, and delivered into mouse by intravenous injection and intramuscular injection, respectively. In addition, pGL3-AFPe-ALBp-Luc was transfected into hepatic and non-hepatic cell lines; pVAX1, pVAX1/S, and pGL3-ALBp-Luc were used as controls. The expression of hepatitis B surface antigen (HBsAg) was observed, and luciferase activity in cells was measured. For plasmid pVAX-AFPe-ALBp-S, the expression of HBsAg was observed in hepatic cell lines, but not in a non-hepatic cell line. Using pVAX-S, the expression of HBsAg was observed in both hepatic and non-hepatic cell lines. In cells expressing pGL3-AFPe-ALBp-Luc, the level of luciferase activity was significantly higher in hepatic cell lines, compared with the non-hepatic cell lines. In addition, the level of luciferase activity in cells expressing pGL3-AFPe-ALBp-Luc was significantly higher than that of pGL3-ALBp-Luc in hepatic cell lines, suggesting that AFPe could enhance target gene expression under the control of ALBp. The expression of HBsAg was detected in mouse liver, but not muscle when using pVAX-AFPe-ALBp-S. In contrast, the expression of HBsAg was detected in both mouse liver and muscle upon transfection with pVAX-S. In conclusion, the AFPe-ALBp element could be used as a tool to induce liver-specific expression of a target gene.

Case Report: A Novel Homozygous Variant Identified in a Chinese Patient With Benign Recurrent Intrahepatic Cholestasis-Type 1.

Benign recurrent intrahepatic cholestasis (BRIC) is a rare hereditary cholestatic liver disorder. Accurate diagnosis and timely interventions are important in determining outcomes. Besides clinical and pathologic diagnosis, genetic study of BRIC remains limited. Here, we report a young man enduring recurrent jaundice and severe pruritus for 15 years. The increased level of direct bilirubin was the main biochemical abnormality, and the work-up for common causes of jaundice were unremarkable. Liver biopsy showed extensive cholestasis of hepatocytes in zone 3. The novel homozygous variant including c.1817T > C and p.I606T was detected on his ATP8B1gene. The patient was finally diagnosed with BRIC-1. His symptoms were relieved, and liver function tests returned to normal after taking ursodeoxycholic acid. This case provides a different perspective to the methodology employed when dealing with cases of jaundice and helping diagnose rare diseases.

Histological changes in chinese chronic hepatitis B patients with ALT lower than two times upper limits of normal.

The role of ALT as a predictor of liver injury has been questioned. The aim of this study is to use liver biopsy to assess the degree of liver injury in patients with chronic hepatitis B(CHB) whose ALT < 2 x upper limit of normal (ULN). A total of 49.2% of patients in this study had significant inflammation (grade >or=2) and 36.4% had significant fibrosis (stage >or=2). The frequency of serious inflammation and fibrosis was similar in patients with different ALT levels. The level of serum HBV DNA was not significantly associated with the extent of inflammation and fibrosis. Advanced age was a significant independent predictor of histological damage and the presence of more significant inflammation and fibrosis. We conclude that many CHB patients with ALT < 2 x ULN have significant liver inflammation or fibrosis and that liver biopsy is necessary to assess liver damage and should be used to assess the need for anti-viral therapy.

Leprosy in a low-incidence setting : Case report relevant to metagenomic next generation sequencing applications.

Leprosy is a disease caused by Mycobacterium leprae that results in disability. In 2000 the World Health Organization announced that leprosy had been eradicated. In nonendemic areas diagnosing leprosy is becoming a challenge for inexperienced clinicians. This case involves a male patient suffering from chronic numbness, hand deformity and recurrent erythema. Skin biopsy revealed granuloma and acid-fast staining of short-rod bacteria. Peripheral venous blood was subjected to metagenomic next generation sequencing and bioinformatics analysis, which revealed 3 unique sequence reads of M. leprae. Paraffin-embedded tissue and fresh samples scraped from skin lesions were subjected to in-house PCR targeting 16S rRNA, hsp65, rpoB, rpoT, ribF-rpsO, and mmaA. Sanger sequencing of amplicons from fresh samples and paraffin-embedded tissue verified the presence of M. leprae. For inexperienced clinicians in nonendemic areas nucleic acid amplification tests, such as in-house PCR, are helpful for diagnosing leprosy but sequence reads from metagenomic next generation sequencing may also provide evidence when interpreted cautiously.

A novel predictive score for citrate accumulation among patients receiving artificial liver support system therapy with regional citrate anticoagulation.

Patients with liver failure may suffer citrate accumulation when using regional citrate anticoagulation for artificial liver support system therapy (RCA-ALSS therapy). This study aimed to develop a predictive scoring system to stratify the risk of citrate accumulation. A total of 338 patients treated with RCA-ALSS therapy were retrospectively enrolled and randomly divided into derivation and validation cohorts. Longer duration of citrate accumulation (LDCA) was defined as the presence of citrate accumulation 2 h after RCA-ALSS therapy. Four baseline variables were found to be independently associated with LDCA: gender, international normalized ratio of prothrombin time, serum creatinine, and serum chloride. A predictive R-CA model and its simplified R-CA score were developed. The R-CA model (AUROC = 0.848) was found to be superior to the MELD score (AUROC = 0.725; p = 0.022) and other univariate predictors (AUROCs < 0.700; all p ≤ 0.001) in predicting LDCA. The R-CA score (AUROC = 0.803) was as capable as the R-CA model (p = 0.369) and the MELD score (p = 0.174), and was superior to other univariate predictors (all p < 0.05) in predicting LDCA. An R-CA score of 0-2 had a negative predictive value of 90.2% for LDCA. Our R-CA score reliably predicts LDCA in patients with RCA-ALSS therapy, and it is easy to use. Patients with R-CA score of 0-2 can safely receive RCA-ALSS therapy, while others should be carefully evaluated before treatment.Trial registration: Chinese Clinical Trial Registry, ChiCTR2000029179. Registered 17 January 2020, https://www.chictr.org.cn/showproj.aspx?proj=48084.