[Intraoperative slide rail CT assistance in percutaneous sacroiliac joint screws for the treatment of pelvic posterior ring injury].

OBJECTIVE: To compare the clinical efficacy of intraoperative slide rail CT combined with C-arm X-ray assistance and just C-arm for percutaneous screw in the treatment of pelvic posterior ring injury. METHODS: A retrospective analysis was performed on the patient data of 76 patients with posterior pelvic ring injury admitted to the Department of Orthopedic Trauma from December 2018 to February 2022. Among them, 39 patients in the CT group were treated with C-arm combined with slide rail CT-assisted inline fixation including 23 males and 16 females with an average age of (44.98±7.33) years old;and the other 37 patients in the C-arm group were treated with intraline fixation treatment under only C-arm fluoroscopy including 24 males and 13 females with an average age of (44.37±10.82) years old. Among them, 42 patients with anterior ring fractures were treated with percutaneous inferior iliac spines with internal fixation (INFIX) or suprapubic support screws to fix the anterior pelvic ring. Postoperative follow-up time, operation time, complications of the two groups were compared. Results of Matta reduction criteria, Majed efficacy evaluation, the CT grading and the rate of secondary surgical revision were compared. RESULTS: The nailing time of (32.63±7.33) min in CT group was shorter than that of (52.95±10.64) min in C-arm group (t=-9.739, P<0.05). The follow-up time between CT group (11.97±1.86) months and C-arm group (12.03±1.71) months were not statistically significant(P>0.05). The postoperative complication rates between two groups were not statistically significant (χ2=0.159, P>0.05). Results of Matta reduction criteria (Z=2.79, P<0.05), Majeed efficacy evaluation(Z=2.79, P<0.05), CT grading (Z=2.83, P<0.05) in CT group were better than those in C-arm group(P<0.05); the secondary surgical revision rate in the CT group was significantly lower than that in the C-arm group (χ2=5.641, P<0.05). CONCLUSION: Compared with traditional C-arm fluoroscopy, intraoperative slide rail CT combined with C-arm assisted percutaneous sacroiliac joint screw placement surgery has the characteristics of short operation time, high accuracy and safety, and significant decrease in postoperative secondary revision rate, and is one of the effective methods for re-establishing the stability of the posterior ring of pelvic fracture.

Silica nanoparticle design for colorectal cancer treatment: Recent progress and clinical potential.

Colorectal cancer (CRC) is the third most common cancer worldwide and the second most common cause of cancer death. Nanotherapies are able to selectively target the delivery of cancer therapeutics, thus improving overall antitumor efficiency and reducing conventional chemotherapy side effects. Mesoporous silica nanoparticles (MSNs) have attracted the attention of many researchers due to their remarkable advantages and biosafety. We offer insights into the recent advances of MSNs in CRC treatment and their potential clinical application value.

Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer.

Exosomes are messengers for intercellular communication and signal transduction. Circular RNA (circRNA) abnormal expression and regulation are involved in the occurrence and development of a variety of tumors. In the present study, exosomes in the serum of five patients with non-small cell lung cancer (NSCLC) were isolated before and after EGFR-TKIs resistance, and the circRNA expression profile was screened using a circRNA microarray. The effects of the exosome circRNA_102481 on cell proliferation and apoptosis were analyzed. The interaction between miR-30a-5p and circRNA_102481 or ROR1 was predicted by starBase software, and was confirmed by RNA pull-down and dual-luciferase reporter assays. The results showed that exosomes containing circRNA_102481 were significantly up-regulated in NSCLC with EGFR-TKIs resistance (p<0.05), and that circRNA_102481 was mainly secreted by EGFR-TKIs resistance cell via exosomes (p<0.05). Both circRNA_102481 silencing and si-circRNA_102481 transported by exosomes could inhibit EGFR-TKIs resistance cell proliferation and promote cell apoptosis and circRNA_102481 overexpression could promote EGFR-TKIs sensitive cell proliferation and inhibit cell apoptosis in vitro (p<0.05). CircRNA_102481 served as a miR-30a-5p sponge to regulate ROR1 expression (p<0.05). Furthermore, the expression of circRNA_102481 in exosomes was associated with TNM stage, tumor differentiation status, brain metastasis, and PFS and OS duration. Therefore, it was concluded that tumor-derived exosomal circRNA_ 102481 could contribute to EGFR-TKIs resistance via the microRNA-30a-5p/ROR1 axis in NSCLC. Exosomal circRNA_102481 may serve as a novel diagnostic biomarker and a therapeutic target for EGFR-TKIs resistance in NSCLC.

[Phenylhexyl isothiocyanate (PHI) regulates histone methylation and acetylation and induces apoptosis in SMMC-7721 cells].

OBJECTIVE: To investigate the effects of PHI on histone acetylation and methylation in hepatocellular carcinoma line SMMC-7721 cells. METHODS: Apoptosis was measured by TUNNEL assay. Histone methylation and acetylation were detected by Western blot. RESULTS: PHI inhibited cells growth and induced apoptosis. PHI treatment resulted in increased acetylation of histone H3 and H4 , elevated level of histone H3 lysine 4 methylation, and decreased level of histone H3 lysine 9 methylation. CONCLUSIONS: PHI can modulate both histone acetylation and methylation, which could remodel chromatin structure. PHI may be a novel anticancer drug.

The calmodulin-binding protein kinase 3 is part of heat-shock signal transduction in Arabidopsis thaliana.

SUMMARY: Based on our previous findings, we proposed a pathway for the participation of Ca(2+)/calmodulin (CaM) in heat-shock (HS) signal transduction. The specific mechanism by which CaM regulates activation of heat-shock transcription factors (HSFs) is not known. CaM-binding protein kinases (CBK) are the most poorly understood of the CaM target proteins in plants. In this study, using a yeast two-hybrid assay, we found that AtCBK3 interacts with AtHSFA1a. Fluorescence resonance energy transfer was used to confirm the interaction between AtCBK3-YFP and AtHSFA1a-CFP. Furthermore, we demonstrate that purified recombinant AtCBK3 phosphorylated recombinant AtHSFA1a in vitro. We also describe the results of both downregulation of AtCBK3 expression and ectopic overexpression in Arabidopsis thaliana. The T-DNA insertion AtCBK3 knockout lines had impaired basal thermotolerance, which could be complemented by transformation of plants with the native gene. Overexpression of AtCBK3 resulted in plants with increased basal thermotolerance. Results from real-time quantitative PCR and protein gel-blot analyses suggest that AtCBK3 regulates transcription of heat-shock protein (HSP) genes and synthesis of HSPs. The binding activity of HSF to the heat-shock element (HSE), the mRNA level of HSP genes and synthesis of HSPs were upregulated in AtCBK3-overexpressing lines after HS, but downregulated in AtCBK3 null lines. These results indicate that AtCBK3 controls the binding activity of HSFs to HSEs by phosphorylation of AtHSFA1a, and is an important component of the HS signal transduction pathway.

A comparison of several second-order algorithms for simultaneous determination of neomangiferin and mangiferin with severe spectral overlapping in Anemarrhenae Rhizoma.

This work presents a greener approach for simultaneous determination of neomangiferin and mangiferin, the major bioactive constituents with severe spectral overlapping in Anemarrhenae Rhizoma, combining the sensitivity of molecular fluorescence and the selectivity of chemometric multivariate calibration algorithms. In this study, we compared the analytical performance of two group chemometric algorithms including trilinear algorithms such as parallel factor analysis (PARAFAC), alternating trilinear decomposition (ATLD), self-weighted alternating trilinear decomposition (SWATLD) and alternating penalized trilinear decomposition (APTLD), and PLS-based methods such as unfolded partial least-squares or the multi-dimensional partial least-squares, both combined with residual bilinearization (U-PLS/RBL, N-PLS/RBL). The statistical parameters for the validation set of the second calibration were evaluated through the relative error of prediction (REP%), the average recovery (Rec%), and the root mean square error of prediction (RMSEP). Prediction results for the validation set by trilinear algorithms showed that the values were satisfactory for neomangiferin, and higher and not acceptable values for mangiferin, while U-PLS and N-PLS predictions were very successful for two analytes. Therefore, U- and N-PLS/RBL were chosen to determine neomangiferin and mangiferin in more complex real samples simultaneously, and U-PLS/RBL algorithm showed the best performance. The predicted concentrations by proposed methods were satisfactorily compared with those obtained using high performance liquid chromatography with ultraviolet detection.

Time-resolved fluorescence and chemometrics-assisted excitation-emission fluorescence for qualitative and quantitative analysis of scopoletin and scopolin in Erycibe obtusifolia Benth.

This paper presents a new strategy for qualitative identification of scopoletin and scopolin in Erycibe obtusifolia Benth using time-resolved (lifetimes) fluorescence and quantitative analysis with chemometrics-assisted excitation-emission matrix (EEM) fluorescence. Due to the significant spectral overlapping among analytes and interference, the use of the more selective time-resolved fluorescence is proposed for qualitative identification in quality control of traditional Chinese medicine (TCM) for the first time. Using the strategy of combining EEM fluorescence with second-order calibration method, i.e. parallel factor analysis (PARAFAC), the simultaneous quantification of scopoletin and scopolin in the complex system of Erycibe obtusifolia Benth was achieved successfully. The predicted concentrations were compared with the values obtained using high performance liquid chromatography-coupled to fluorimetric detector (HPLC-FLD), and no significant differences between them were observed. Therefore, the proposed methods using time-resolved fluorescence for qualitative analysis and EEMs coupled with second-order calibration for quantitative analysis in TCM are comparable and provide a suitable alternative to the chromatography-based method.

Simultaneous determination of α-asarone and ß-asarone in Acorus tatarinowii using excitation-emission matrix fluorescence coupled with chemometrics methods.

A chemometrics-assisted excitation-emission matrix (EEM) fluorescence method was proposed for simultaneous determination of α-asarone and ß-asarone in Acorus tatarinowii. Using the strategy of combining EEM data with chemometrics methods, the simultaneous determination of α-asarone and ß-asarone in the complex Traditional Chinese medicine system was achieved successfully, even in the presence of unexpected interferents. The physical or chemical separation step was avoided due to the use of "mathematical separation". Six second-order calibration methods were used including parallel factor analysis (PARAFAC), alternating trilinear decomposition (ATLD), alternating penalty trilinear decomposition (APTLD), self-weighted alternating trilinear decomposition (SWATLD), the unfolded partial least-squares (U-PLS) and multidimensional partial least-squares (N-PLS) with residual bilinearization (RBL). In addition, HPLC method was developed to further validate the presented strategy. Consequently, for the validation samples, the analytical results obtained by six second-order calibration methods were almost accurate. But for the Acorus tatarinowii samples, the results indicated a slightly better predictive ability of N-PLS/RBL procedure over other methods.

Clinical observations on the treatment of prolapsing hemorrhoids with tissue selecting therapy.

AIM: To compare the effects and postoperative complications between tissue selecting therapy stapler (TST) and Milligan-Morgan hemorrhoidectomy (M-M). METHODS: Four hundred and eighty patients with severe prolapsing hemorrhoids, who were admitted to the Shenyang Coloproctology Hospital between 2009 and 2012, were randomly divided into observation (n=240) and control (n=240) groups. Hemorrhoidectomies were performed with TST in the observation group and with the M-M technique in the control group. The therapeutic effects, operation security, and postoperative complications in the two groups were compared. The immediate and long-term complications were assessed according to corresponding criteria. Pain was assessed on a visual analogue scale. The efficacy was assessed by specialized criteria. The follow-up was conducted one year after the operation. RESULTS: The total effective rates of the observation and control groups were 99.5% (217/218) and 98.6% (218/221) respectively; the difference was not statistically significant (P=0.322). Their were significant differences between observation and control groups in intraoperative blood loss (5.07±1.14 vs 2.45±0.57, P=0.000), pain (12 h after the surgery: 5.08±1.62 vs 7.19±2.01, P=0.000; at first dressing change: 2.64±0.87 vs 4.34±1.15, P=0.000; first defecation: 3.91±1.47 vs 5.63±1.98, P=0.001), urine retention (n=22 vs n=47, P=0.001), anal pendant expansion after the surgery (2.35±0.56 vs 5.16±1.42, P=0.000), operation time (18.3±5.6 min vs 29.5±8.2 min, P=0.000), and the length of hospital stay (5.3±0.6 d vs 11.4±1.8 d, P=0.000). Moreover TST showed significant reductions compared to M-M in the rates of long-term complications such as fecal incontinence (n=3 vs n=16, P=0.003), difficult bowel movement (n=1 vs n=9, P=0.011), intractable pain (n=2 vs n=12, P=0.007), and anal discharge (n=3 vs n=23, P=0.000). CONCLUSION: TST for severe prolapsing hemorrhoids is a satisfactory technique for more rapid recovery, lower complication rates, and higher operation security.

[Fluorescence of Tb(3+) -calmodulin complex and its analytical application].

Calmodulin (CaM) is a ubiquitous Ca(2+) -binding protein of eukaryotes, and regulates a broad spectrum of fundamental cellular processes. CaM harbors four binding domains, among which domains I, II and III contain no tyrosine, only domain IV has one tyrosine for plant species. In contrast to mammals, plants express numerous CaM isoforms that exhibit differential activation or inhibition of CaM dependent enzymes in vitro. In the present study, the isoform II of Arabidopsis thaliana CaM was used to test the binding properties of metal ion to CaM by Tb3+ fluorescence. The increase in fluorescence was monitored at 545 nm as a function of the number of Tb3+ bound to CaM using direct (221 nm) and indirect (280 nm ) excitation. Upon direct excitation of Tb(3+) -CaM, the fluorescence of Tb3+ increased markedly, and one of the pathways of energy dissipation of the excited state of Tb3+ was energy transfer to the vibrational levels of water molecules in the hydration sphere around the Tb3+ ion. When waters of hydration were removed as a result of Tb3+ binding to CaM, an increase in rate constants of luminescence was observed. The titration curve with direct excitation increased up to 4 mol of Tb3+/mol of CaM before the onset of a plateau, in agreement with the expected maximum of four binding sites. Using indirect excitation at 280 nm, the resultant titration curve was sigmoid, albeit with less fluorescence intensity, also reached a maximum at a ratio of 4 mol of Tb3+/mol of CaM, in which the first phase exhibits an end at 2 : 1, and in this phase there was only a small increase in Tb3+ fluorescence. The fact that only the second pair of added Tb3+ shows a large enhancement in Tb3+ fluorescence suggests that it is Tb3+ bound to the low affinity sites that can accept energy from tyrosine group. The turning points of fluorescence titration curves were used to estimate the concentrations of CaM on the base of CTb3+ = 4c(CaM).

[Anatomical and clinical study of the supinator syndrome evoked embitterment test].

OBJECTIVE: To explore the mechanism and feasibility of the supinator syndrome evoked embitterment test from anatomy and clinic. METHODS: 25 cases of The supinator syndrome were reviewed. 18 of them were male and 7 were female. Drop finger deformation were apparent in 25 cases and The supinator syndrome evoked embitterment test was positive for All patients. Operative neurolysis was done in 8 cases, conservation treatment 17 cases; 92 cadaver upper extremities were dissected for a study the relationship between supinator tunnel and posterior interosseous nerve. RESULTS: 22 cases had been followed up for an average of 9 months. 16 cases had a full recovery and 6 cases, a partial recovery. the anatomical study shows that The posterior interosseous nerve was compressed by Forhse arcade and the distal border of the supinator muscle during passive pronation forearm. CONCLUSION: The supinator syndrome evoked embitterment test was a new test for the diagnosis of supinator syndrome, it was found to be more sensitive and specific than the others test.

[Preliminary investigation of coblation for early glottic carcinoma].

OBJECTIVE: To establish whether Coblation is a suitable modality for removal of early glottic carcinoma. METHODS: Fourteen patients with early glottic carcinoma (Tis-T2) without lymph node metastasis underwent resection of laryngeal cancer lesions using transoral endoscopic coblation (TEC), without pre- or post-operative radiotherapy and chemotherapy. RESULTS: No severe complication such as bleeding and dyspnea occurred in the cases. Only mild postoperative pain happened to the patients. All patients could eat and pronounce on the surgery day. With following-up of 25 - 37 months, no recurrence was observed in 13 cases. One case, with poorly differentiated squamous cell lesion in the anterior commissure invading subglottic, recurred 3 months postoperatively. The patient received the re-resection of laryngeal lesion by coblation, but another recurrence happened to the patient 6 months postoperatively, and then was cured by partial laryngectomy, with recurrence-free survival 21 months postoperatively. CONCLUSION: The observation suggests that transoral endoscopic coblation is a reliable and safe modality for the resection of early glottic carcinoma.

[Effect of phenylhexyl isothiocyanate on adriamycin resistance of K562/A02 cell line].

This study was to investigate the effect of phenylhexyl isothiocyanate (PHI) on drug resistance and sensitivity on K562/A02 cell line to adriamycin (ADM) and to elucidate the possible mechanisms. The inhibition rates of ADM and PHI + ADM on growth of K562/A02 cell line were measured by MTT assay, and K562/A02 cell resistant multiple was calculated. The apoptosis rate of K562/A02 cell line, the changes of intracellular ADM concentrations and MRP1 protein level were detected by flow cytometry (FCM). Intracellular reoxidized GSH level was determined by spectrometric enzyme assay and MRP1 mRNA was assayed by semiquantitative RT-PCR before and after using PHI. The results indicated that the survival rate of K562/A02 cell line decreased with the increasing concentration of PHI. Apoptosis rate increased after treatment in combination with two above drugs, the changes of drug resistance multiple and intracellular ADM level had statistical significance between K562/A02 and K562 cells (p < 0.05), when the concentration of PHI was more than 20 micromol/L. Intracellular GSH level in K562/A02 cell line reduced 5% when 1 microg/ml ADM was used alone, and it increased slightly at first, then decreased when more than 10 micromol/L PHI was used. When more than 20 micromol/L PHI was used in combination with 1 microg/ml ADM, intracellular GSH level in K562/A02 cell line decreased progressively with increasing the concentration of PHI. The expressions of MRP1 mRNA and protein had no statistical significance between K562/A02 and K562 cells (p > 0.05) after or before PHI was used. It is concluded that the cyto-toxicity of PHI to K562/A02 cell line does not associate with the depletion of the intracellular GSH. PHI not only enhances the sensitivity of K562/A02 cell line to ADM, but also partially reverses effect of K562/A02 cell resistance to ADM. ADM combined with PHI can diminish side effect and dosage of ADM.

[Effect of phenylhexyl isothiocyanate on inducing apoptosis of multiple myeloma cells in vitro].

In order to investigate the effects of phenylhexyl isothiocyanate (PHI) on proliferation and apoptosis of multiple myeloma cells RPMI8226 in vitro, the RMPI8226 cells were co-cultured with PHI at various concentrations; the apoptosis of PHI-treated cells was assayed by TUNEL; the cell cycle changes of PHI-treated cells were analyzed by FCM; the mitochondrial potential changes of PHI-treated cells were detected by using a potential sensitive dye JC-1 as probe; the VEGF levels secreted from PHI-treated cells were measured by quantitative sandwich ELISA. The results showed that PHI significantly inhibited RPMI8226 cell proliferation, induced their apoptosis at low concentration (0.5 micromol/L), the inhibitory effect was related to PHI concentration. PHI-treated cells were arrested in G(0)/G(1) phase. The RPMI8226 cells showed shift from red fluorescence to green fluorescence in some concentration-dependent manner, indicating increase of mitochondrial depolarization and potential loss by 3-4-fold as compared with control, after treated RMPI8226 cells with 10 micromol/L of PHI for 48 hours, the VEGF level secreted from RMPI8226 cells significantly decreased, it was 35% of control. It is concluded that the PHI can inhibit cell proliferation, induce cell apoptosis of RMPI8226, the cell apoptosis is associated with mitochondrial depolarization and potential loss, the inhibiting VEGF secretion from RMPI8226 cells by PHI may be one of the reasons causing apoptosis. PHI may be a potential therapeutic drug for multiple myeloma.

[Advance of study on demethylation in leukemia - review].

DNA methylation is a frequent change in epigenetics of leukemia. In the development of leukemia, methylation plays an important role in the genetic expression regulation and genetic structure stabilization. Hematopoietic stem cell transplantation (HSCT) and chemotherapy are the common methods to cure leukemia at present. Unfortunately, it is very hard to find suitable donor for transplantation, while the relapse and drug resistance are the unresolved problems in chemotherapy. Because of the reversibility of methylation, the drugs reducing the level of methylation become a new way to cure leukemia. Decitabine is the most common medicament used to reduce the level of methylation. In this article, the methylation and tumor, methylation and hematologic diseases, methylation detection methods, demethylation therapy in leukemia and so on are reviewed.

[Phenylhexyl isothiocyanate reducing U266 cell line methylation level of p16 gene].

This study was purposed to investigate whether phenylhexyl isothiocyanate (PHI) can reduce p16 gene methylation level or not. The myeloma U226 cell line was cultured with PHI of 0, 5, 10 micromol/L for 72 hours, then DNA was extracted. Hydrosulfite was used to treat the genome DNA of healthy adult, PCR amplification was carried out by using this DNA as template. The gene chip detecting methylation changes of 3 CpG in promoter region of p16 gene was constructed by designing a pair of probes which contain one methylated and one unmethylated probes. This pair of probes was used to detect 3 consecutive sites of CpG island in p16 gene. The standard curve was constructed by using gene chip after the methylated and unmethylated DNA were mixed at different ratio. Then treated samples of U266 cells were dotted on gene chip, obtained results were compared with standard curve to get the quantitative results. The results indicated that the probes on chip had excellent reproducible ability and precision, the methylation level of p16 gene in U266 cells treated with 0, 5 and 10 micromol/L of PHI was determined as 78.2%, 61.7% and 54.8%, respectively. It is concluded that the PHI can reduce the methylation level of p16 gene in U266 cells.

[Experiment study of PHI on histone methylation and acetylation in Molt-4 cells].

OBJECTIVE: To investigate the effect of phenyl-hexyl isothiocyanate (PHI) on acetylation and methylation of histone in acute lymphoblastic leukemia cell line Molt4. METHODS: The inhibition of cell proliferation was observed by MTT method and clone suppression test. Apoptosis and cell cycle arrest were measured by flow cytometry. The alterations in histone acetyltransferase and acetylation and methylation of histones were detected by Western blot. RESULTS: PHI could up-regulate the expression of acetyltransferase (P300/CBP), markedly induced the accumulation of acetylated histone H3, H4 and methylated histone H3 lysine 4 (H3K4), and inhibited methylation on lysine 9 of H3 (H3K9). The epigenetic regulation resulted in cell cycle arrest at G0/G1 phase, and induction of apoptosis. CONCLUSIONS: PHI can modulate both histone methylation and acetylation. It may serve as a histone deacetylase inhibitor, and might be a potential novel anti-leukemia agent.

[Surgical treatment of tumors in anterior and middle skull base by modified maxillary bone disassembly procedures].

OBJECTIVE: To explore the therapeutic effect of the modified maxillary bone disassembly procedures on patients with tumors in the anterior and middle skull base. METHODS: Retrospective analysis was made of patients treated with the modified maxillary bone disassembly procedures. Ten tumors in the anterior and middle skull base were resected according to the pathology, size and site of the skull base tumors including 9 benign tumors and 1 malignant tumor. RESULTS: All tumors were resected completely. All patients were followed up from 12 months to 5 years postoperatively. The complications were less and the life quality was increased. The patients with benign tumors showed no recurrence . Two patients with cerebrospinal rhinorrhea cured spontaneously after 7 days. One patient with melanoma died of brain metastase. CONCLUSIONS: It is necessary to estimate the tumors thoroughly before surgery. According to the location of the tumor, the modified maxillary bone disassembly is the nearest and harmless approach, through which the tumors can be completely excised with minimal invasiveness. The life quality is thus increased.