Identification of potential biomarkers of vascular calcification using bioinformatics analysis and validation in vivo.

Background: Vascular calcification (VC) is the most widespread pathological change in diseases of the vascular system. However, we know poorly about the molecular mechanisms and effective therapeutic approaches of VC. Methods: The VC dataset, GSE146638, was downloaded from the Gene Expression Omnibus (GEO) database. Using the edgeR package to screen Differentially expressed genes (DEGs). Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were used to find pathways affecting VC. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed on the DEGs. Meanwhile, using the String database and Cytoscape software to construct protein-protein interaction (PPI) networks and identify hub genes with the highest module scores. Correlation analysis was performed for hub genes. Receiver operating characteristic (ROC) curves, expression level analysis, GSEA, and subcellular localization were performed for each hub gene. Expression of hub genes in normal and calcified vascular tissues was verified by quantitative reverse transcription PCR (RT-qPCR) and immunohistochemistry (IHC) experiments. The hub gene-related miRNA-mRNA and TF-mRNA networks were constructed and functionally enriched for analysis. Finally, the DGIdb database was utilized to search for alternative drugs targeting VC hub genes. Results: By comparing the genes with normal vessels, there were 64 DEGs in mildly calcified vessels and 650 DEGs in severely calcified vessels. Spp1, Sost, Col1a1, Fn1, and Ibsp were central in the progression of the entire VC by the MCODE plug-in. These hub genes are primarily enriched in ossification, extracellular matrix, and ECM-receptor interactions. Expression level results showed that Spp1, Sost, Ibsp, and Fn1 were significantly highly expressed in VC, and Col1a1 was incredibly low. RT-qPCR and IHC validation results were consistent with bioinformatic analysis. We found multiple pathways of hub genes acting in VC and identified 16 targeting drugs. Conclusions: This study perfected the molecular regulatory mechanism of VC. Our results indicated that Spp1, Sost, Col1a1, Fn1, and Ibsp could be potential novel biomarkers for VC and promising therapeutic targets.

Identification of potential biomarkers for diagnosis of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) has a high mortality rate owing to its complexity. Identification of abnormally expressed genes in HCC tissues compared to those in normal liver tissues is a viable strategy for investigating the mechanisms of HCC tumorigenesis and progression as a means of developing novel treatments. A significant advantage of the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) is that the data therein were collected from different independent researchers and may be integrated, allowing for a more robust data analysis. Accordingly, in the present study, the gene expression profiles for HCC and control samples were downloaded from the GEO and TCGA. Functional enrichment analysis was performed using a Metascape dataset, and a protein-protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of Interacting Genes/proteins (STRING) online database. The prognostic value of mRNA for HCC was assessed using the Kaplan-Meier Plotter, a public online tool. A gene mRNA heatmap and DNA amplification numbers were obtained from cBioPortal. A total of 2,553 upregulated genes were identified. Functional enrichment analysis revealed that these differentially expressed genes (DEGs) were mainly accumulated in metabolism of RNA and the cell cycle. Considering the complexity and heterogeneity of the molecular alterations in HCC, multiple genes for the prognostication of patients with HCC are more reliable than a single gene. Thus, the PPI network and univariate Cox regression analysis were applied to screen candidate genes (small nuclear ribonucleoprotein polypeptide B and B1, nucleoporin 37, Rac GTPase activating protein 1, kinesin family member 20A, minichromosome maintenance 10 replication initiation factor, ubiquitin conjugating enzyme E2 C and hyaluronan mediated motility receptor) that are associated with the overall survival and progression-free survival of patients with HCC. In conclusion, the present study identified a set of genes that are associated with overall survival and progression-free survival of patients with HCC, providing valuable information for the prognosis of HCC.

Efficacy and safety of transjugular intrahepatic portosystemic shunt combined with transcatheter embolization/chemoembolization in hepatocellular carcinoma with portal hypertension and arterioportal shunt.

OBJECTIVES: This study seeks to assess the efficacy and safety of transjugular intrahepatic portosystemic shunt (TIPS) combined with transarterial embolization/transarterial chemoembolization (TAE/TACE) in hepatocellular carcinoma (HCC) with portal hypertension and arterioportal shunt (APS). METHODS: Consecutive hospitalized patients having HCC accompanied by portal hypertension and APS were retrospectively analyzed. A total of 103 patients were enrolled. Of them, 26 patients were in Group A and 77 patients were in Group B according to the treatment protocol (Group A: TIPS plus TAE/TACE; Group B: TAE/TACE alone). The clinical outcomes and survival rate were compared between the two groups. RESULTS: The mean survival time in Group A and Group B were 14 mo and 9.9 mo, respectively, with statistical difference (p = 0.043). The immediate APS improvement rate was 95.2% in Group A and 91.9% in Group B, respectively, with no signficant difference (p = 1.000). However, the first follow-up consultation revealed that APS improvement rate in Group A was more obvious (66.7% vs 27.4%, p = 0.001). Objective response rate of HCC tended to be greater in Group A compared with Group B (65.4% vs 38.7%, p = 0.019). Liver function parameters significantly increased in Group A than those in Group B. After TIPS placement, the mean portal pressure gradient decreased from 32.61 ± 8.87 mmHg to 15.61 ± 8.15 mmHg, with significant difference (p = 0.000). The rate of absorption of ascites and control of variceal bleeding were statistically different between the two groups (p = 0.045 and 0.039, respectively). CONCLUSION: Our research suggests that TIPS combined with TAE/TACE seems to be safe and efficacious in patients with HCC accompanied by portal hypertension and APS, albeit may be accompanied by liver function damage.

Multidisciplinary treatment of advanced hepatocellular carcinoma with severe arterioportal shunt: a case report.

Hepatocellular carcinoma (HCC) is the fourth leading cause of cancer mortality globally. In addition, most patients present in advanced stages with limited curative treatment options. Therefore, multidisciplinary treatment is often warranted. Here, we report a patient with HCC and severe arterioportal shunt (APS) who was treated with a multidisciplinary approach comprising interventional radiology procedures, apatinib and camrelizumab. After treatment, the intrahepatic mass was stable, and a notable decrease in the number and size of lung lesions was observed. The patient achieved a long-term survival of more than 2 years. These data suggest that multidisciplinary treatments may be effective in the treatment of advanced HCC with severe APS.

Potentially Inappropriate Medication among Older Patients Who Are Frequent Users of Outpatient Services.

Aging is accompanied by changes in organ degeneration, and susceptibility to multiple diseases, leading to the frequent occurrence of adverse drug reactions resulting from polypharmacy (PP) and potentially inappropriate medications (PIM) in older patients. This study employs a retrospective cohort design and investigates the association of PP with PIM among older patients with high rates of medical utilization. Using records from a national pharmaceutical care database, an experimental group is formed from patients meeting these criteria, who are then offered home pharmaceutical care. Correspondingly, a control group is formed by identifying older patients with regular levels of use of medical services who had been dispensed medications at community pharmacies. Multivariate logistic regression is performed to assess the association between the rate of PIM and variables, including age, gender, and PP. The study finds that experimental PP participants had a higher rate of PIM prescription (odds ratio (OR) = 5.4) than non-PP control participants (all p < 0.001). In clinical practice, additional caution is required to avoid PIMs. Patients engaged in continuously using long-term medication should take precautions in daily life to alleviate related discomforts. Pharmacists should serve as a bridge between patients and physicians to enhance their health and improve their quality of life.

MAPRE1 promotes cell cycle progression of hepatocellular carcinoma cells by interacting with CDK2.

Targeting cyclin-dependent kinases (CDKs) is a promising method of therapy for cancer. Unfortunately, the efficacy of CDK inhibitors in hepatocellular carcinoma (HCC) is limited, due in part to incomplete understanding of cell cycle progression and a lack of specific biomarkers to adequately identify which patients may be responsive to CDK inhibitors. In the present study, we report that microtubule-associated protein RP/EB family member 1 (MAPRE1), a gene involved in cell cycle and microtubule regulation, is significantly increased in HCC tissue, promotes HCC cell proliferation, enhances in vitro tumorigenesis, and associates with poor prognosis of HCC. We demonstrate that MAPRE1 binds with CDK2, resulting in the hyperphosphorylation of the CDK2 Thr161 residue in HCC cells. Our findings reveal that targeting MAPRE1 might be an effective therapeutic strategy in HCC, and suggest that MAPRE1 expression might provide a promising biomarker to stratify patients with HCC who may benefit from treatment with CDK inhibitors.

Effect of Pharmacist Intervention on a Population in Taiwan with High Healthcare Utilization and Excessive Polypharmacy.

Patients with high healthcare utilization are at increased risk of polypharmacy and drug interactions. This study investigated the changes in the number of medications, drug interactions and interaction severity in high frequency outpatients with polypharmacy at hospitals and clinics in Taiwan after home pharmaceutical care, to understand the effectiveness of interventions by pharmacists. This was a retrospective observational study. Cases with excessive polypharmacy (10+ drugs) were selected from the Pharmaceutical Care Practice System database of the Taiwan Pharmacist Association in 2017. After the home care intervention, the number of drug types used decreased 1.89-fold (p < 0.001), and the number of medications fell 61.6%. The incidence of drug interaction was 93.82%. In an average case, the incidence of drug interaction after the pharmacist intervention decreased 0.6-fold (p < 0.001). The drug most commonly causing interactions was aspirin, followed by diclofenac; also common were three used in diabetes, two psycholeptics and two beta blockers. Among 22 cases of severe drug interaction, seven resulted in increased risk of extrapyramidal symptoms and neuroleptic malignant syndrome. By analyzing the relationship between the side effects of individual drugs and the pharmacokinetic Tmax, a sequential thermal zone model of adverse drug reactions can be established, the value of which could prompt physicians and pharmacists to intervene in order to prevent adverse events. It is concluded that home pharmaceutical care by pharmacists can significantly reduce the number of medications and interactions in patients with excessive polypharmacy and high healthcare utilization.

PPFIA1 is upregulated in liver metastasis of breast cancer and is a potential poor prognostic indicator of metastatic relapse.

Although the oncogenic role of PPFIA1 (liprin-α1) in breast cancer has been reported, whether its dysregulation is associated with metastasis risk or survival outcomes in breast cancer patients is not clear. Our primary data showed that PPFIA1 expression was significantly higher in liver metastatic breast tumors than in the primary tumors. Then, we tried to pool previous annotated genomic data to assess the prognostic value of PPFIA1 in distant metastasis-free survival, the risk of metastatic relapse, and metastatic relapse-free survival in breast cancer patients by data mining in two large databases, Kaplan-Meier plotter and bc-GenExMiner 4.0. Results from Kaplan-Meier plotter showed that although high PPFIA1 expression was generally associated with decreased distant metastasis-free survival in estrogen receptor+ patients, subgroup analysis only confirmed significant association in estrogen receptor+/N- (nodal negative) group (median survival, high PPFIA1 group vs low PPFIA1 cohort: 191.21 vs 236.22 months; hazard ratio: 2.23, 95% confidence interval: 1.42-3.5, p < 0.001), but not in estrogen receptor+/N+ (nodal positive) group (hazard ratio: 1.63, 95% confidence interval: 0.88-3.03, p = 0.12). In estrogen receptor- patients, there was no association between PPFIA1 expression and distant metastasis-free survival, no matter in Nm (nodal status mixed), N-, or N+ subgroups. In bc-GenExMiner 4.0, Nottingham Prognostic Index- and Adjuvant! Online-adjusted analysis validated the independent prognostic value of PPFIA1 in metastatic risks in estrogen receptor+/N- patients. Based on these findings, we infer that high PPFIA1 expression might be an independent prognostic indicator of increased metastatic relapse risk in patients with estrogen receptor+/N- breast cancer, but not in estrogen receptor+/N+ or estrogen receptor- patients.

MicroRNA-30e reduces cell growth and enhances drug sensitivity to gefitinib in lung carcinoma.

MicroRNAs (miRNAs) play critical roles in variousbiological processes,including malignancy. Here, we demonstrated that miR-30e levels were markedly reduced in human lung carcinoma specimens in comparisonwith adjacent normal tissues.In addition, miR-30eamounts were starkly lower in the resistant PC9/gefitinib (PC9G) cancer cells compared with PC9 cells. Meanwhile, miR-30eoverexpression inPC9G cells resulted in reduced cell proliferation and migration,reversing drug resistance to gefitinib.Conversely,miR-30e silencing in PC9 cells increased proliferation as well as migration, and conferred resistance to gefitinib.Moreover, HOXA1, which was identified asa new miR-30etarget, plays important roles in regulating cell fate, early developmental patterns and organogenesis.Importantly, miR-30ealso inhibited PC9G growth in vivo. Taken together, these findings demonstrated that miR-30eshould be considered a tumor suppressor miRNA, which could be used in treatinghuman lung cancer.

[Retrospective study of complication of interspinous implants for degenerative lumbar disease].

OBJECTIVE: To summarize and analyze the complications of interspinous implants for degenerative lumbar disease. METHODS: From September 2007 to September 2011, 177 cases with degenerative lumbar diseases were treated with interspinous implants. There were 99 male patients and 78 female patients, the average age was 44.5 years (26 - 71 years). According to the application interspinous dynamic stabilization system type were divided into the Wallis group (136 cases) and Coflex group (41 cases). The clinical results were assessed by visual analog scale (VAS) of pain on lumbar and lower limbers, lumbar Japanese Orthopedic Association (JOA) score and Prolo functional score. The radiological results including segmental lodosis and segement movement degree were assessed by lumbar X ray and dynamic X ray. Summarize and analyze the complications both during operation and post operation. Quantitative datas were compared by paried-samples t test and complication rate was compared by χ(2) test. RESULTS: There were 168 cases had completed follow-up and the average time was 34.7 months (3 - 50 months). In the final follow-up, lumbar pain VAS, lower limber pain VAS, lumbar JOA score and Prolo functional score were better than pre-operation (t = 10.7, 7.9, 13.4 and 8.8, P < 0.01). Segment lodosis angles was 14° ± 4° which was less than pre-operation 19° ± 4° (t = 9.4, P < 0.01).Segment movement degree was larger in Coflex group (12.6° ± 3.1°) than in Wallis group (9.7° ± 2.7°) (t = 8.6, P < 0.05). Complication rate was 10.7% (18/168), which of Wallis group was 6.2% (8/130) and Coflex group was 26.3% (10/38) (χ(2) = 12.5, P < 0.01). In Wallis group, there were 3 cases with dura tear and cerebrospinal fluid leakage, 1 case with nerve root injury and foot drop, 2 cases with spacer breakage when implantation and change the implants and 2 cases with recurrence of lumbar disc herniation. In Coflex group, there was 1 case with dura tear and cerebrospinal fluid leakage, 2 cases with mild displacement post operation, 1 case with debridement for aseptic wound exudates, 1 case with implant removal for breakage 1 week post operation, 4 cases with recurrence of lumbar disc herniation and 1 case with lumbar disc herniation 6 months post operation of lumbar stenosis. CONCLUSIONS: The application of interspinous implants for degenerative lumbar diseases is effective and relative safe, but would suffer from the risk of complications.

[Clinical characteristics and reoperation of symptomatic adjacent segment degeneration postoperation of lumbar fusion].

OBJECTIVE: To evaluate the clinical characteristics and reoperation of symptomatic adjacent segment degeneration postoperation of lumbar fusion. METHODS: In the study, 28 cases of symptomatic adjacent segment degeneration postoperation of lumbar fusion from May 2007 to April 2012 were retrospectively reviewed,with an average age of (64.3±8.7) years. The mean period between reoperation and primary fusion surgery was (47.5±30.8) months. Symptomatic adjacent segment degeneration located in cephalic segments in 12 cases, in caudal segments in 14 cases and in both segments in 2 cases. Of all the 28 patients, 12 suffered from lumbar stenosis, other 13 from lumbar disc herniation and still other 3 from both lumbar stenosis and disc herniation, of whom 11 were together with segment instability. Localization diagnoses of 19 cases were clear and other 9 received diagnostic nerve root block. The cases were divided into two groups by the type of reoperation,of which 8 cases were in adjacent degenerative segment nonfusion group and the other 20 cases in adjacent degenerative segment fusion group. The clinical results were assessed by lumbar pain visual analog score (VAS) and lower limber pain VAS,lumbar Japanese Orthopedic Association (JOA) score and Prolo functional score before operation and in the final follow-up. RESULTS: In adjacent degenerative segment nonfusion group, the average operation time was (86.3±17.1) min and average blood volume was (125.0 ±37.8) mL of reoperation and 1 case with dural injury; and in adjacent degenerative segment fusion group, the average operation time was (201.6 ±71.0) min and average blood volume was (313.6±218.9) mL of revision surgery and 4 cases with dural injury. The average follow-up period was 25.5 months. In the final follow-up,lumbar pain VAS,lower limber pain VAS,lumbar JOA score and Prolo functional score of adjacent degenerative segment nonfusion group were 2.4±1.2, 2.8±1.4,23.5±4.2 and 8.2±1.5,which were better than preoperation 5.5±2.9, 6.8±2.5, 13.7±5.2 and 4.3±2.1, P<0.001; lumbar pain VAS, lower limber pain VAS,lumbar JOA score and Prolo functional score of adjacent degenerative segment nonfusion group were 3.3±1.9, 3.1±1.2, 22.2±4.4 and 7.7±1.6, which were better than preoperation 5.4±2.7, 7.0±2.4, 13.0±5.6 and 3.9±1.9, P<0.001.In the final follow up, lumbar pain VAS of adjacent degenerative segment nonfusion group was better than that adjacent degenerative segment fusion group(P=0.028). CONCLUSION: Symptomatic adjacent segment degeneration postoperation of lumbar fusion is difficult for diagnosis and treatment. Appropriate reoperation could get the good results.

[Prevention for venous thromboembolism prophylaxis after spinal surgery].

OBJECTIVE: To investigate the efficacy and safety of venous thrombus embolism (VTE) prophylaxis according to risk stratifications after spinal surgery. METHODS: From June 2008 to June 2009, we separated 298 spinal patients who had different VTE risk factors into low-, medium- and high-risk groups for 22 cases, 48 cases and 228 cases respectively. Physical prevention measures such as thigh-length thromboembolic deterrent stockings (TEDS) and pneumatic sequential compression device (PSCD) were used in low- and medium-risk groups. In high-risk groups, low molecular weight heparin(LMWH) was applied in addition to physical prevention measures. Lower limb vascular doppler ultrasonography was used to monitor thrombosis pre- and postoperatively. Simultaneously the occurrences of epidural or wound hematoma, mucosal bleeding, thrombocytopenia caused by low molecular heparin and nerve damage caused by extradural hemorrhage were monitored. RESULTS: Among the 298 cases of patients with spinal surgery, DVT occurred in 23 cases, the incidence of DVT was 7.7%. There were 0, 2 and 21 patients with positive findings of deep vein thrombosis on duplex ultrasonograph respectively in low-, medium- and high-risk groups. There was no case of PE. All DVT was thrombosis in calf which was distal to the knee. There was no clinical symptom of VTE. The DVT needed no therapy. The vein with thrombosis was recanalized 3 months after operation. No case caught epidural or wound hematoma, mucosal bleeding, thrombocytopenia caused by low molecular heparin or nerve damage caused by extradural hemorrhage. CONCLUSION: Individual VTE prophylaxis was taken according to risk stratifications. No VTE of clinical value or no complications from prophylaxis happened. So our prophylaxis is effective and safe. But more prospective, case-control studies are needed to assess the efficacy and safety of VTE prophylaxis.

[Retrospective study of combined application of interspinous process fixation system and rigid fixation system for degenerative lumbar diseases].

OBJECTIVE: To assess the efficacy and safety of combined application of interspinous process fixation system and rigid fixation system for degenerative lumbar diseases. METHODS: From September 2007 to September 2008, 16 cases with degenerative lumbar diseases were treated with combined application of interspinous process fixation system and rigid fixation system. The clinical results were assessed by VAS of pain of lumbar and lower limbs, lumbar JOA score and Prolo functional score. The radiological results including implant position (interspinous process, pedicle screws and plates), spinous process fracture, segmental range of motion (the non-fusion fixation segment, and the intermediate segments between fused and non-fused segments) which were assessed by lumbar static and dynamic X rays. RESULTS: All 16 cases obtained an average follow-up of 17.6 months. At final follow up, lumbar VAS, lower limbers VAS, lumbar JOA score and Prolo functional score were significant improved than those of pre-operation (lumbar VAS: 1.9 +/- 1.4 vs. 4.5 +/- 3.1; lower limbs VAS: 1.7 +/- 1.2 vs. 6.3 +/- 2.9; lumbar JOA score: 22.8 +/- 3.3 vs. 12.5 +/- 4.7; Prolo functional score: 8.3 +/- 1.2 vs. 4.0 +/- 2.3). Range of motion of the non-fusion fixation segment was (9.8 +/- 4.2) degrees and that of the intermediate segments between fused and non-fused segments was (13.2 +/- 3.5) degrees . CONCLUSIONS: Combined application of interspinous process fixation system and rigid fixation system for degenerative lumbar diseases provides a new idea to avoid the multi-segment fusion fixation and pertinent potential problems. Short-term clinical results are successful.

[Effect of pedicle screw fixation with bone cement in lumbar].

OBJECTIVE: To evaluate the strength of pedicle screw fixation with bone cement in human cadaveric lumbar vertebral bodies by using biomechanical test. METHODS: Twenty-four fresh frozen cadaveric lumbar vertebral bodies were randomly divided into 3 groups, which each group had 8 bodies. The bodies in the first group were drilled only. The bodies in the second group were drilled with tap. The bodies in the third group were drilled and the holes were enlarged by borer. Each vertebral body was drilled in both right and left sides, which one side served as control with the pedicle screws fixation only and another side served as testing side with the pedicle screw fixation with bone cement. The pullout strength was tested on both sides by using borer. RESULT: There was significant difference of manifestation of pull-strength between the control side and the testing side in all groups (P < 0.001). For the manifestation of pull-strength of testing side in different groups, which had pedicle screw fixation with bone cement, there was no significant difference between the second group and third group, however, the difference between the first group and the second group and between the first group and third group were significant. CONCLUSION: The pedicle screws with bone cements could significantly enhance the strength of fixation in cadaveric lumbar compared with pedicle screws only.