Characteristics and ecological risks of microplastic pollution in a tropical drinking water source reservoir in Hainan province, China.

Microplastic (<5 mm) pollution has become a pressing environmental concern in recent years. The present study investigated the occurrence characteristics and assessed the ecological risk of microplastics in the surface water and sediment of the Chitian Reservoir, a drinking water source in Hainan province (China). The results indicated that microplastics were detected in the surface water and sediment of the Chitian Reservoir and its surrounding areas. The overall abundance of microplastics in the water was 3.05 ± 1.16 items per L and in the sediment was 0.15 ± 0.06 items per g dry weight, which is relatively low compared to other reservoirs in China. The dominant components of microplastics detected in the Chitian Reservoir were polypropylene (PP), rayon, and polyester. Physical morphology analysis of microplastics showed that fibers with small particle sizes (<1 mm) and white color were the predominant characteristics in both the surface water and sediment. The domestic sewage from surrounding residents and agricultural wastewater may be the primary sources of microplastics in the reservoir. Ecological risk assessment revealed that the overall pollution load index (PLI) in the surface water (0.65) and sediment (0.51) of the Chitian Reservoir and its surrounding area is at a low level. The potential ecological hazards (RI) of microplastics (0.13 to 336.78 in water; 0.23 to 465.93 in sediment) in most sites fall within the scope of level I, but those in a few sites are at level II due to the presence of polyvinyl chloride (PVC). This study enriches the data on microplastic pollution in inland reservoir systems, providing fundamental reference information for future ecotoxicological studies and the management of microplastic pollution control.

Effect of structural stability of lipase in acetonitrile on its catalytic activity in EGCG esterification reaction: FTIR and MD simulation.

In this study, (-)-Epigallocatechin-3-O-gallate (EGCG) esterification reaction was catalyzed by Novozym 435, Lipozyme RM, Lipozyme TLIM, and lipase Amano 30SD in acetonitrile. Fourier transform infrared spectroscopy (FTIR) and molecular dynamic (MD) simulations were used to analyze the structural stability of different lipases in acetonitrile and their effect on EGCG esterification reaction. The results showed that conversion rate of EGCG catalyzed by Lipozyme RM was the highest, followed by Lipozyme TLIM. FTIR indicated that the secondary structure of Lipozyme RM was the most stable. MD simulations suggested that whole structural stability of Lipozyme RM in acetonitrile was superior to Novozym 435 and lipase Amano 30SD and similar to Lipozyme TLIM due to their similar conformation, while the active site of Lipozyme RM is more flexible than that of Lipozyme TLIM, which indicated that lipase with stable whole structure and flexible active site may be more conducive to the esterification of EGCG in acetonitrile. This study provided a direction for rapidly screening lipase to synthetize EGCG or other polyphenols esterified derivatives.

[Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein].

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.

Preparation, identification, and inhibitory mechanism of dipeptidyl peptidase IV inhibitory peptides from goat milk whey protein.

This study explores potential hypoglycemic mechanisms by preparing and identifying novel dipeptidyl peptidase IV (DPP-IV) inhibitory peptides from goat milk (GM) whey protein. Papain was used to hydrolyze the GM whey protein. After purification by ultrafiltration, the Sephadex column, and preparative RP-HPLC, the peptide inhibited DPP-IV, α-glucosidase, and α-amylase with IC50 of 0.34, 0.37, and 0.72 mg/mL, respectively. To further explore the inhibitory mechanism of peptides on DPP-IV, SPPEFLR, LDADGSY, YPVEPFT, and FNPTY were identified and synthesized for the first time, with IC50 values of 56.22, 52.16, 175.7, and 62.32 µM, respectively. Molecular docking and dynamics results show that SPPEFLR, LDADGSY, and FNPTY bind more tightly to the active pocket of DPP-IV, which was consistent with the in vitro activity. Furthermore, the first three N-terminals of SPPEFLR and FNPTY peptides exhibit proline characteristics and competitively inhibit DPP-IV. Notably, the first N-terminal leucine of LDADGSY may play a key role in inhibiting DPP-IV.

[Mechanism of n-butanol alcohol extract of Baitouweng Decoction in treatment of vulvovaginal candidiasis based on negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis].

This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1ß, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1ß, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1ß, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.

Nanosized niosomes as effective delivery device to improve the stability and bioaccessibility of goat milk whey protein peptide.

This study sought to develop a nanoscale delivery system to enhance the stability and bioaccessibility of goat milk whey protein peptides. Goat milk whey protein was hydrolyzed by papain, and the hydrolysate was ultrafiltered to obtain a low molecular weight peptide (GWP, <3 kDa) with strong hypoglycemic activity. The GWP-loaded liposomes and niosomes encapsulation systems were prepared using phytosterols (ergosterol, ß-sitosterol, mixed phytosterols, and stigmasterol) instead of cholesterol. Results showed that the GWP-loaded niosomes (GWP-NS) prepared from ß-sitosterol had the higher GWP encapsulation efficiency (90.46 ± 4.02 %) and the smaller particle size (92.07 ± 9.8 nm) than liposomes (GWP-LS). Additionally, the morphological results showed that two GWP-loaded systems were smooth and spherical, and the FT-IR spectroscopy confirmed the successful formation of the peptide-loaded system. Compared with GWP, GWP-LS and GWP-NS showed the higher stability under different pH, temperature, and NaCl concentration conditions, especially GWP-NS. Furthermore, GWP-NS could significantly improve the retention rate of GWP during simulated gastrointestinal digestion, in vitro bioaccessibility, and hypoglycemic activity. These findings suggest that ß-sitosterol could be a potential membrane stabilizer alternative to cholesterol, and GWP niosomes could be a potential new drug delivery system.

[Risk factors of hidden blood loss in percutaneous vertebral augmentation].

OBJECTIVE: To explore the risk factors of hidden blood loss in osteoporosis vertebral compression fractures during percutaneous vertebral augmentation. METHODS: From October 2018 to December 2019, 360 patients with osteoporosis vertebral compression fractures who received percutaneous vertebral augmentation were enrolled in this study. The factors analyzed included gender, age, surgical methods, disease course, height, weight, the operative segment, bone mineral density, amount of bone cement, operative time, percentage of height loss, percentage of vertebral height restoration, cement leakage, blood clotting function, preoperative and postoperative hemoglobin and hematocrit and other internal diseases. Total blood loss was calculated by Gross's formula, influential factors of the hidden blood loss were further analyzed by t-test, multivariate linear regression and one-way ANOVA analysis. RESULTS: Surgical methods, the operative segment, disease course, cement leakage, preoperative hemoglobin, cement leakage via the basivertebral and segmental vein were significantly correlated with hidden blood loss(P<0.05). CONCLUSION: Patients with percutaneous kyphoplasty, two-level and multi-level surgery, the course of the disease beyond 6 weeks, cement leakage via the basivertebral and segmental vein, and lower preoperative hemoglobin had more perioperative hidden blood loss.

Long-term efficacy and safety of bone cement-augmented pedicle screw fixation for stage III Kümmell disease.

This study aimed to evaluate the efficacy and safety of bone cement-augmented pedicle screw fixation for stage III Kümmell disease. Twenty-five patients with stage III Kümmell disease who received bone cement-augmented pedicle screw fixation at the First Affiliated Hospital of Guangzhou University of Chinese Medicine between June 2009 and December 2015 were enrolled. All patients were females with a history of osteoporosis. The vertebral Cobb angle (V-Cobb angle), the fixed segment Cobb Angle (S-Cobb angle), pelvic parameters, visual Analogue Scale (VAS) score, and Oswestry Disability Index (ODI) were assessed preoperatively, postoperatively and at the final follow-up. Complications, loosening rate, operation time, and intraoperative bleeding were recorded. The average lumbar vertebral density T-value was - 3.68 ± 0.71 SD, and the average age was 71.84 ± 5.39. The V-Cobb angle, S-Cobb angle, and Sagittal Vertical Axis (SVA) were significantly smaller postoperatively compared to the preoperative values. The VAS and ODI at 1 month after surgery were 3.60 ± 1.00 and 36.04 ± 6.12%, respectively, which were both significantly lower than before surgery (VAS: 8.56 ± 1.04, ODI: 77.80 ± 6.57%). Bone cement-augmented pedicle screw fixation is a safe and effective treatment for stage III Kümmell disease. It can effectively correct kyphosis, restore and maintain sagittal balance, and maintain spinal stability.

Comparison of Cement Leakage Rate and Severity After Percutaneous Vertebroplasty for Osteoporotic Vertebral Compression Fractures Using Front-Opening Versus Side-Opening Cannulas.

The authors assessed the occurrence and severity of cement leakage (CL) following percutaneous vertebroplasty (PVP) for osteoporotic vertebral compression fractures (OVCFs) performed using front-opening cannulas (FOCs) vs side-opening cannulas (SOCs). This retrospective cohort study included 811 patients with single-level OVCFs who underwent PVP between March 2016 and September 2018. The 264 patients who met the inclusion criteria were divided into two groups according to whether the procedure was performed using a FOC (n=128) or a SOC (n=136). Visual analog scale score, Oswestry Disability Index, local kyphotic angle, vertebral height, amount of bone cement injected, and rate of CL were compared between the groups. Types of CL were classified according to postoperative computed tomography. The CL types were further classified according to severity (mild and severe). Visual analog scale score, Oswestry Disability Index, local kyphotic angle, and vertebral height were all significantly improved after surgery. The total incidence of CL was significantly higher for FOCs than for SOCs (P=.001). Similarly, the occurrence of the CL subtypes was significantly higher for FOCs than for SOCs (B type, P=.033; C type, P=.01; and S type, P=.015). Analysis of CL type severity revealed that the rates of severe D type (intradiskal leakage category of the C type) and severe S type were significantly lower for SOCs than for FOCs (P=.001), while the incidence of severe B type leaks was not significantly different between the groups (P=.443). Percutaneous vertebroplasty performed using SOCs decreases the incidence and severity of D type and S type CL compared with FOCs. Secondary classification of CL could help physicians better understand the severity of leakage and select a more appropriate surgical approach to reduce CL. [Orthopedics. 2021;44(3):134-140.].

Tubeimoside-I sensitizes colorectal cancer cells to chemotherapy by inducing ROS-mediated impaired autophagolysosomes accumulation.

BACKGROUND: Tubeimoside-I (TBM), a plant-derived bioactive compound, shows antitumor activity in different tumors and can enhance the efficacy of chemotherapeutic agents. However, the detail mechanism underlying remains to be elucidated. METHODS: The cytotoxic potential of TBM towards CRC cells was examined by CCK8 assay, colony formation, LDH release assay, flow cytometry method and Western blots. The ROS levels, autophagy, apoptosis, chemosensitivity to 5-FU or DOX, etc. were determined between control and TBM-treated CRC cells. RESULTS: In this study, we found that TBM could inhibit proliferation and induce apoptosis in colorectal cancer (CRC) cells. Intriguingly, TBM treatment could either promote autophagy initiation by ROS-induced AMPK activation, or block autophagy flux through inhibiting lysosomal hydrolytic enzymes, which leaded to massive impaired autophagylysosomes accumulation. Administration of autophagy initiation inhibitor (3-MA or selective ablation of autophagy related proteins) relieves TBM-induced CRC suppression, while combination use of autophagy flux inhibitor chloroquine (CQ) slightly augments TBM-induced cell death, suggesting that impaired autophagylysosomes accumulation contributes to TBM-induced growth inhibition in CRC cells. Notably, as an autophagy flux inhibitor, TBM works synergistically with 5-fluorouracil (5-FU) or doxorubicin (DOX) in CRC suppression. CONCLUSION: Together, our study provides new insights regarding the anti-tumor activity of TBM against CRC, and established potential applications of TBM for CRC combination therapies in clinic.

[Establish mouse osteoblast -osteoclast cell co-culture system in a Transwell chamber].

OBJECTIVE: To establish osteoblast-osteoclast cell co-culture system in a Transwell chamber, and detect cell viability of osteoblasts and osteoclasts in system. METHODS: Osteoblast MC3T3-E1 and mouse monocytes RAW264.7 were cultivated in vitro. RANKL-induced mouse RAW264.7 monocytes differentiated into mature osteoclasts, osteoblast-osteoclast cell co-culture system was established in Transwell chamber. Cell activity of osteoblasts and osteoclasts were detected by CCK-8 experimenting, Alizarin Red staining, TRAP staining. The expression of OPG, ALP, RANKL, TGF-b1 gene and RANKL protein in osteoblast MC3T3-E1 were detected by PCR, Western-Blot methods. Also, the expression of RANK, NF-κB in gene and protein level in osteoclast were measured through the same method respectively. RESULTS: The co-culture system of Mouse MC3T3-E1 cells and RAW264.7 cell were established in Transwell chamber. Co-culture system affected cell division activities of osteoblasts and osteoclasts. Differentiation of osteoblasts were increased, while differentiation of osteoclast division were slight decreased under microscope observation. OPG (0.65±0.08) and ALP (0.16±0.01) gene expression of co-culture system were less than single culture OPG(1.00±0.08) and ALP (1.01±0.16); TGF-b1(4.42±0.21) and RANKL(4.12±1.04) of osteoblasts in co-culture system were higher than TGF-b1(1.00±0.10) and RANKL(1.00±0.09) under single culture. However, gene expression of RANK(0.63±0.06) and NF-κB(0.64±0.08) in co-culture system were decreased than RANK(1.00±0.08) and NF-κB(1.00±0.09), in single culture, and had significant differences. Similarly, protein expression of OPG(0.43±0.05) and NF-κB(0.59±0.05) of co-culture system were less than OPG(0.84±0.06) and NF-κB(1.13±0.03) of single culture. While RANKL protein expression (0.54±0.03)of co-culture system was more than single culture RANKL(0.31±0.03), and had statistically differences, which was in agreement of the trend of gene expression change. CONCLUSIONS: Co-culture system of mouse MC3T3-E1 cells and RAW264.7 cell was viable in Transwell chamber, and the activity of osteoblasts is higher than osteoclasts in co-culture system.

Risk Factors for the Occurrence of Insufficient Cement Distribution in the Fractured Area after Percutaneous Vertebroplasty in Osteoporotic Vertebral Compression Fractures.

BACKGROUND: Insufficient cement distribution (ICD) in the fractured area has been advocated to be responsible for unsatisfied pain relief after percutaneous vertebroplasty (PVP) for osteoporotic vertebral compression fractures (OVCFs). However, little is known about risk factors for the occurrence of ICD. OBJECTIVE: The present study aimed to identify independent risk factors of the emergence of ICD. STUDY DESIGN: A retrospective cohort study. SETTING: Department of spinal surgery, an affiliated hospital of a medical university. METHODS: Patients who underwent PVP for single-level OVCF from January 2012 to September 2014 and met this study's inclusion criteria were retrospectively reviewed. Associations of ICD with co-variates (age, gender, bone mass density with a T-score, amount of injected cement, cement leakage, fracture level, fracture age, fracture severity grade, and location of the fractured area) and the influence of ICD on pain relief were analyzed. RESULTS: A total of 225 patients were included. ICD was found in 26 (11.6%) patients. Fractured area located in the superior portion of the index vertebra was significantly associated with occurrence of ICD. No further significant associations between the studied co-variates and emergence of ICD were seen in the adjusted analysis. In addition, patients with ICD had significantly higher immediate postoperative visual analog scale scores of back pain compared with those with sufficient cement distribution in the fractured area. LIMITATION: Location of the fractured area and cement distribution in the fractured area could not be evaluated quantitatively. CONCLUSIONS: The incidence of ICD is higher in patients with the fractured area located in the superior portion of the index vertebra and ICD might be responsible for unsatisfied pain relief after PVP for OVCFs. KEY WORDS: Percutaneous vertebroplasty, insufficient cement distribution, fractured area, risk factor, osteoporosis, vertebral compression fracture, spine, unsatisfied pain relief, cement augmentation.

Unique expression signatures of circular RNAs in response to DNA tumor virus SV40 infection.

Circular RNAs (circRNAs), identified as a class of widely expressed endogenous regulatory RNAs, are involved in diverse physiological and pathological processes. However, their role in viral pathogenesis and cellular antiviral response remains unexplored. In this study, a potent DNA tumor virus, simian virus 40 (SV40), was used as a model to investigate the viral influences on cellular circRNA transcriptome. Using RNA-seq, 15,241 putative circRNAs were identified de novo from 5,057 parental genes in monkey kidney-derived Vero cells. The expression of selected circRNAs was confirmed by reverse transcription-polymerase chain reaction and Sanger sequencing. Further analysis showed that most circRNAs comprised multiple exons, and most parental genes produced multiple circRNA isoforms. A total of 134 significantly dysregulated circRNAs, including 103 upregulated and 31 downregulated circRNAs, were found after SV40 infection. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that various cancer-related pathways, including p53 and Wnt pathway, could be affected by SV40 infection via the alteration of the circRNA hosting genes. Moreover, diverse cellular immune pathways, including Toll-like receptor pathway and Janus kinase-signal transducer and activator of transcription pathway, could also be affected by SV40 infection. An integrated circRNA-miRNA-gene analysis suggested the putative function of circRNAs as cellular and viral miRNA decoys to indirectly regulate the gene expression during SV40 infection. This study presented the first comprehensive expression and functional profile of circRNAs in response to SV40 infection, thus providing new insights into the mechanisms of viral pathogenesis and cellular immune response.

Icariin influences adipogenic differentiation of stem cells affected by osteoblast-osteoclast co-culture and clinical research adipogenic.

OBJECTIVE: To build mouse osteoblast MC3T3-E1 and mouse osteoclast RAW264.7 co-culture system and to study the effect of icariin on the activity of osteoblasts and osteoclasts in the co-culture system. METHODS: In vitro acquisition and cultivation of mouse osteoblasts MC3T3-E1 and mouse RAW264.7 cells were conducted. Osteoblast and osteoclast activities of cells were detected by CCK-8 staining experiment, alizarin red staining and tartaric-resistant acid phosphatase (TRAP) staining. We used different concentrations of icariin to interfere in osteoblast-osteoclast co-culture system. The effects of icariin on various genes were detected by PCR and Western blot methods The correction between the expression of PPARγ and BMD was analyzed in patients with osteoporosis. RESULTS: Mouse osteoblast-osteoclast co-culture system was built, and the osteogenic differentiation effect was enhanced. Icariin can improve the MC3T3-E1 osteogenic differentiation activity, enhance the expression of OPG and RANKL gene protein, reduce the NF-κb gene and protein expression, increase of ALP, TGF-b1 and RANKL gene expression level and reduce RANK gene expression. Icariin can act on MC3T3-E1 cells-RAW264.7 cells co-culture system, and promote the osteogenic activity of MC3T3-E1 cells, inhibit the osteoclast activity of RAW264.7 cells and reduce the level of BMSCs adipogenic differentiation. The expression level of PPAR-γ gene was negatively correlated with the level of BMD. CONCLUSIONS: Mouse MC3T3-E1 cells and mouse RAW264.7 cells could be co-cultured in vitro, and icariin could improve the osteogenic activity of MC3T3 cells-RAW264.7 cells and decrease the osteoclast activity. Icariin could inhibit adipogenic differentiation of BMSCs in the osteoblast-osteoclast co-culture, promoting osteogenic differentiation and inhibiting osteoclast differentiation.

[CLASSIFICATION AND TREATMENT STRATEGIES OF SYMPTOMATIC SEVERE OSTEOPOROTIC VERTEBRAL FRACTURE AND COLLAPSE].

OBJECTIVE: To investigate the classification and treatment strategies of symptomatic severe osteoporotic vertebral fracture and collapse. METHODS: Between August 2010 and January 2014, 42 patients with symptomatic severe osteoporotic vertebral fracture and collapse were treated, and the clinical data were retrospectively analyzed. According to clinical symptom and imaging materials, 23 cases were classified as type I (local pain, limitation of motion, no neurological symptom, and no obvious deformity), 12 cases as type II (slight neurological symptom and kyphotic Cobb angle < or = 30 degrees), and 7 cases as type III (severe neurological symptom and kyphotic Cobb angle >30 degrees). In 23 type I patients, 17 underwent percutaneous vertebral augmentation, 6 underwent posterior pedicle screw fixation strengthened with bone cement combined with percutaneous vertebral augmentation. In 12 type II patients, they were treated with local spinal decompression and internal fixation strengthened with bone cement. In 7 type III patients, 5 underwent posterior osteotomy, and 2 underwent one stage posterior approach of vertebral resection and reconstruction. The visual analogue scale (VAS), Oswestry disability index (ODI), and local kyphotic Cobb angle were used to evaluate the neurological function. The complications were recorded. RESULTS: The operation was successfully completed in all patients. Wound infection and ketoacidosis secondary to stress blood glucose rise occurred in 1 case of type III patients respectively, and were cured after corresponding treatment; primary healing of wound was obtained in the other patients. The patients were followed up from 6 to 36 months (mean, 11.6 months). The nerve function was improved in 17 cases, and micturition disability was observed in 2 cases. Asymptomatic cement leakage occurred in 13 cases (30.95%) (7 cases in type I, 4 cases in type II, and 2 cases in type III). No bone cement dislocation and internal fixation failure were found during follow-up. The VAS score, ODI, and the local kyphotic Cobb angle at 1 week and last follow-up were significantly improved when compared with preoperative ones (P<0.05), but no significant difference was found between at 1 week and last follow-up (P>0.05). CONCLUSION: In order to improve the effectiveness and reduce the risk and complications of operation, individualized strategies should be performed according to different types of severe osteoporotic vertebral fracture and collapse.

Predicting the Binding Affinity of ERß Ligands Based on a Novel Variable Selection Method.

A number of descriptors were employed to characterize the molecular structures of the 128 estrogen receptor ß ligands. A quantitative structure-activity relationship (QSAR) model of these compounds was developed by the variable selection method based on variable interaction. The QSAR model with five descriptors was internally and externally validated. The determination coefficient (R 2) and the leave-one-out cross-validated correlation coefficient (Q 2) are 0.8272 and 0.8041, respectively. The estimated correlation coefficient of the external validation is 0.8255. The mechanistic interpretation of the final model was carried out according to the definition of descriptors. As the model meets the five principles proposed by Organization for Economic Co-operation and Development, it can be used to predict the binding affinity of other derivatives.

Pap Smear Combined with HPV Testing: A Reasonable Tool for Women with High-grade Cervical Intraepithelial Neoplasia Treated by LEEP.

BACKGROUND: To evaluate HPV testing by Hybrid Capture II (HCII) in conjunction with cytology in detecting the residual/recurrence disease after treatment of high-grade cervical intraepithelial neoplasia (CIN II-III) with loop electrosurgical excision procedure (LEEP). MATERIALS AND METHODS: A retrospective review of 158 patients with histologically confirmed CIN II-III who underwent LEEP between January 2011 and October 2012 was conducted. Post-treatment control was scheduled at the 3rd, 6th, 12th and 18th month. All patients were followed up by Pap smear and HR-HPV genotype and viral load testing. RESULTS: Pre-treatment, HR-HPV DNA, was detected in all specimens of the patients. At follow-up, 25 patients were diagnosed as the residual/recurrent disease during the FU visit, among whom, 16 patients with positive margin: 13 patients (52%) with HR-HPV DNA+/cytology+, 2 patients (8%) with HR-HPV DNA+/cytology-, 1 patient (4%) with cytology+/ HR-HPV DNA-; 9 patients with clean margin--5 patients (55.6%) with HR-HPV DNA+/cytology+; 2 patients (22.2%) with HR- HPV DNA+/cytology-, 2 patients (22.2%) with cytology+/HR-HPV DNA-. None of them persisting HR-HPV DNA-/cytology- with positive or negative margin was identified as the residual/recurrent disease. The majority of residual/recurrent disease was detected at the 12th and 18th month FU, and there was almost no difference in the sensitivity and negative predictive value (NPV) between at the 3rd month and the 6th month FU visits. 14 residual/recurrence disease (14/46:30.4%) had pre-treatment high viral load (>5,000 RUL/PC) and 11 (11/112, 9.8%) with pre-treatment low viral load, P<0.05. CONCLUSIONS: (1) The persistence HR-HPV DNA is the root cause of the residual/recurrent disease for the women treated for high-grade CIN; the pre-treatment viral load and margin can be seen as the predictor. (2) The FU visit beginning at the 6th month post-treatment and lasting at least 24 months with the combination of cytology and HPV testing. (3) Patients with high pre-treatment HPV load, which is considered as one risk of developing the residual/recurrent disease, should be paid more attention (especially above 500 RUL/PC) to by clinicians.

[COMPARISON OF EFFECTIVENESS AND RADIOLOGICAL FEATURES BETWEEN SINGLE AND DOUBLE CAGE IMPLANTING THROUGH UNILATERAL TRANSFORAMINAL LUMBAR INTERBODY FUSION].

OBJECTIVE: To compare the effectiveness and imaging features between implanting single and double Cage into intervertebral body through unilateral transforaminal lumbar interbody fusion (TLIF). METHODS: The clinical data were collected and analyzed from 104 patients who underwent unilateral TLIF between January 2013 and October 2014, who were divided into 2 groups: single Cage was implanted into intervertebral body in 64 cases (76 segments) in traditional group, and double Cage was implanted into intervertebral body in 56 cases (70 segments) in reformative group. There was no significant difference in age, gender, bone mineral density, operation segments between 2 groups (P > 0.05). The visual analogue scale (VAS), Oswestry disability index (ODI), and Japanese Orthopedic Association (JOA) scores were used to evaluate the effectiveness; the area of intervertebral bone-graft, fusion rate, height of intervertebral space, and the number of Cage subsidence were measured by CT scan. RESULTS: All the patients were followed up 12.85 months on average (range, 9-15 months). The VAS, ODI, and JOA scores were significantly improved at each time point after operation when compared with preoperative values (P < 0.05), and no significant difference was found between 2 groups (P > 0.05) except VAS and ODI at 12 months after operation (P < 0.05). However, the area of intervertebral bone-graft in reformative group [(5.94 ± 1.17) cm2] was significantly larger than that in traditional group [(4.81± 0.97) cm2] at 7 days after operation (t = -6.365, P = 0.000). At 3 and 12 months after operation, the fusion rate was respectively 84.2% and 92.1% in traditional group and was respectively 88.6% and 94.3% in reformative group. Although the height of intervertebral space were increased when compared with preoperative height, the incidence rates of Cages subsidence in traditional group were 44.74% and 47.37% respectively at 3 and 12 months after operation and were significantly higher those that in reformative group (11.43% and 14.29% respectively) (P < 0.05). In addition, the height difference between affected side and normal side in traditional group was significantly larger than that in reformative group (P < 0.05). CONCLUSION: Both single and double Cage implanted into the intervertebral body through unilateral TLIF have good effectiveness. However, double Cage implanted into intervertebral body may hold the height of intervertebral space, reduce the incident rate of Cage subsidence, and prevent sagittal imbalance.

Oxidized casein impairs antioxidant defense system and induces hepatic and renal injury in mice.

SCOPE: Oxidized protein products (OPPs) can be easily found in meat and milk during processing and storage. Evidence supports that accumulation of endogenous OPPs plays a negative role in physiological metabolism. However, the impacts of dietary OPPs and the mechanisms have not been elucidated yet. The present study evaluated whether oral oxidized casein would destruct the antioxidant defense system and cause potential oxidized injury in mice liver and kidney. METHODS AND RESULTS: We performed oxidized casein (modified respectively by H2O2-Cu and HClO) feeding experiments using KM mice (20-22 g). A 10-weeks feeding of oxidized casein as basal protein caused oxidative stress by increasing protein carbonylation (PC), advanced oxidation protein products (AOPPs), dityrosine (Dityr), lipid peroxidation and ROS levels in mice liver, kidney and blood (P<0.05). In mice liver and kidney, the mRNA expression of Nrf2, γ-GCS, HO-1, GPX-3, and GPX-4 up-regulated, the protein level of Nrf2 in nucleus increased. However, activities of anti-oxidant enzymes (CAT, SOD, and GPX) decreased (P<0.05). Moreover, histopathological examination displayed the formation of fibrous septa in mice liver and kidney after oxidized casein feeding. CONCLUSION: Oxidized casein impairs antioxidant defense system and induces hepatic and renal fibrosis.

[Effectiveness of cochlear implantation in children with auditory neuropathy and cochlear nerve aplasia].

OBJECTIVE: The aim of this study is to assess of cochlear implantation in children with auditory neuropathy and cochlear nerve aplasia by using Categories of Auditory Performance (CAP) and Speech Intelligibility Rating (SIR). METHODS: Twenty one children with cochlear implants participated in this study. They all received cochlear implant surgery at our hospital from January 2004 to October 2010. All children had hearing aid trial and hearing and speech rehabilitation before surgery at least three months.Nine children (7 male, 2 female) were diagnosed with auditory neuropathy, twelve (7 male, 5 female) with cochlear nerve aplasia. Twenty children (10 male, 10 female) with sensorineural hearing loss served as a control group. All the children received cochlear implant for more than six months. Forty two children with normal hearing served as another control group which were divided into three subgroups according to their age.Group A included 18 children aged under two yrs, group B consisted of 16 children aged from two to four yrs and group C comprised eight children aged above four yrs. CAP and SIR were used to evaluate among all the children and the scores were compared. RESULTS: The CAP scores of children with auditory neuropathy, cochlear nerve aplasia, sensorial neural hearing loss and the three subgroups children with normal hearing were 4.44 ± 1.50, 4.83 ± 1.69, 4.55 ± 1.66, 5.22 ± 1.11, 6.75 ± 0.45 and 7.00 ± 0.00 respectively, and SIR scores were 2.66 ± 1.11, 2.33 ± 1.15, 2.40 ± 0.75, 2.56 ± 1.04, 4.12 ± 0.81 and 5.00 ± 0.00 respectively. There were significant differences among the six groups for CAP scores(χ(2) = 35.481, P < 0.001) and SIR scores(χ(2) = 40.549, P < 0.001).No significant differences for CAP and SIR scores were observed between children with auditory neuropathy/cochlear nerve aplasia and sensorial neural hearing loss as well as group A (P > 0.05 for each), and there were significant differences were shown between children with auditory neuropathy/cochlear nerve aplasia and group B as well as group C (P < 0.01 for each aplasia). CONCLUSIONS: The auditory and speech capabilities of children with auditory neuropathy and cochlear nerve deficiency can can get benefits from cochlear implants as children with sensorineural hearing loss, however not achieve the level of those with normal hearing after cochlear implantation. The long term effects still need follow-up and evaluation.