Objective: To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell (PBMC) recovery with high viability, long-term storage stability, and sterility of the harvested residues, thereby providing a clinically translatable strategy. Methods: Three sterile bag-packaged solutions—Saline, Solvent, and Hanks' balanced salt solution (HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters. Nucleated cell recovery rate and viability of the harvested residues were compared. The optimal solution identified was applied to an expanded sample set. PBMC viability and yield were evaluated after 1h vs 48h storage of the residues. PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month, followed by post-thaw comparisons of viability and T-cell expansion capacity. Results: The Solvent group achieved the highest and most consistent nucleated cell recovery rate. Post-flush recovery rate from filters after 400 mL whole blood processing was (21.3±1.6)% for the Solvent group, significantly higher than Saline group (19.2±6.3)% and HBSS group (11.2±5.0)%, with residues from all groups maintaining viability >90%. No biologically significant difference in residue viability was observed between 48h vs 1h storage groups (93.3±2.3)% vs (95.7±1.8)%). PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups [(48.2%±9.5%)vs (40.41%±8.35%), P>0.05], with (17.7±2.6)×10 cells. After 1-month cryopreservation and 10-day expansion, PBMCs isolated from 48-hour-stored residues retained (91.2±3.2)% viability and achieved a (61.9±15.9)-fold expansion. Conclusion: The bag-packaged Solvent, as a back-flush solution, enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections. These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃, with post-cryopreservation (1-month liquid nitrogen) viability and expansion capacity remaining stable. This protocol complies with blood bank regulatory criteria, addresses the concerns about the infectious window period in cell therapy raw materials, and provides a clinically translatable strategy for PBMC-based applications.

Objective To investigate the effect of parecoxib pretreatment on focal cerebral ischemia-reperfusion (I/R) injury in rats. Methods Sixty-four male SD rats weighing 250-300 g were randomly divided into 4 groups ( n= 16 each): sham operation group (group S); focal cerebral I/R group; focal cerebral I/R + parecoxib 5 mg/kg group (group P5); focal cerebral I/R + parecoxib 10 mg/kg group (group P10). Focal cerebral I/R was produced by occlusion of middle cerebral artery for 2 h followed by 24 h of reperfusion. Parecoxib 5 and 10 mg/kg were injected intravenously through the internal jugular vein 30 min before ischemia in group P5 and P10 respectively. The neurologic deficit scores (NDSs) were measured at 24 h of reperfusion and then the rats were decapitated.Brains were rapidly removed for determination of the infarct volume, apoptosis rate and expression of Bcl-2 and Bax. The ratio of Bcl-2 to Bax (Bcl-2/Bax) was calculated. Results The NDSs, apoptosis rate and expression of Bcl-2 and Bax were significantly higher, Bcl-2/Bax was significantly lower, and the infarct volume was significantly larger in group I/R than in group S ( P ＜ 0.01 ). The NDSs were significantly lower in group P10, and the apoptosis rate and Bax expression were significantly lower, the infarct volume was significantly smaller, Bcl-2 expression and Bcl-2/Bax were significantly higher in group P5 and P10 than in group I/R (P ＜0.05 or 0.01). The infarct volume was significantly smaller, the apoptosis rate and Bax expression were significantly lower, and Bcl-2 expression and Bcl-2/Bax were significantly higher in group P10 than in group P5 ( P ＜ 0.05 or 0.01 ). Conclusion Pretreatment with parecoxib can attenuate focal cerebral I/R injury in a dose-dependent manner through inhibition of cell apoptosis via up-regulation of Bcl-2 expression and down-regulation of Bax expression in rats.

OBJECTIVETo evaluate the efficacy and toxicity of concurrent chemoradiotherapy for patients with locally advanced unresectable extrahepatic cholangiocarcinoma.

METHODSThirty-eight patients with locally advanced unresectable extrahepatic cholangiocarcinoma admitted in Shaoxing People's Hospital from February 2007 to February 2012 were enrolled in the study. They were randomized into sequential chemoradiotherapy (n=19) or concurrent chemoradiotherapy group (n=19). All patients were treated with intensity modulated radiation therapy (IMRT). Patients in concurrent chemoradiotherapy group received the regimen of gemcitabine plus oxaliplatin. Tumor response and adverse effects were observed periodically. The primary end points were disease progression-free survival (PFS) and overall survival (OS).

RESULTSThe response rates of sequential chemoradiotherapy and concurrent chemoradiotherapy groups were 42.1% (8/19) and 63.2% (12/19). The disease control rates of them were 78.9% (15/19) and 84.2% (16/19))， respectively. The median PFS of sequential chemoradiotherapy group and concurrent chemoradiotherapy group was 8.3 (95%CI: 7.6-9.0) and 10.4 months (95%CI: 9.4-11.4, P=0.037), and the median OS in two groups were 14.2 (95%CI: 12.6-15.8) and 15.6 months (95%CI: 14.2-17.0, P=0.095), respectively. The major adverse reactions were controllable hematology toxicity and gastrointestinal reaction. There was no significant difference in incidence of adverse reactions between two groups (P>0.05).

CONCLUSIONSequential chemoradiotherapy and concurrent chemoradiotherapy may improve PFS and OS in patients with locally advanced unresectable extrahepatic cholangiocarcinoma, and both are well-tolerated. In addition, concurrent chemoradiotherapy might provide additional PFS benefit and would be preferable.

Bile Duct Neoplasms ; therapy ; Bile Ducts, Intrahepatic ; pathology ; Chemoradiotherapy ; Cholangiocarcinoma ; therapy ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Humans ; Organoplatinum Compounds ; therapeutic use ; Survival Rate

Objective: To investigate the clinical effect of orthodontic-repair combined therapy in the treatment of maxillary anterior teeth dentition defect and malocclusion. Methods: The clinical data of 68 patients with maxillary anterior dentition defect and malocclusion were analyzed retrospectively.They were divided into control group (repair treatment) and study group (orthodontic-repair treatment) according to the different methods of treatment, 34 cases in each group.The tooth occlusal relationship, the occlusion of the anterior teeth and the arrangement of the teeth, clinical treatment effect and the treatment cost were analyzed.The satisfaction of the patients after treatment was investigated by questionnaire.The debris index and attachment loss index were detected. Results: The good rate of tooth occlusal relationship, good rate of anterior teeth coverage and teeth order rate in the study group were 82.35%(28/34), 88.24%(30/34), 94.12%(32/34), respectively, which were significantly higher than those in the control group[58.82%(20/34), 67.65%(23/34), 73.53%(25/34)](χ²=4.53, 4.19, 4.53, P=0.03, 0.04, 0.03). The total effective rate of the study group was 94.12%(32/34), which was significantly higher than that of the control group[70.59%(24/34)](χ²=6.50, P=0.03). The average treatment cost of the study group was (8 124.63±386.58)yuan, which was significantly higher than (2 179.86±276.82)yuan of the control group (t=72.90, P=0.00). The patients' satisfaction of the study group was was 88.24%(30/34), which was significantly higher than that of the control group[52.94%(18/34)](χ²=10.20, P=0.00). The debris index and attachment loss index in the study group were (0.14±0.07), (5.06±1.38), respectively, which were significantly lower than those in the control group[(0.63±0.18), (6.52±1.58)](t=14.79, 14.79, P=0.00, 0.00). Conclusion Orthodontics combined with orthodontic treatment can help to improve the patients' tooth deformity, and the effect is remarkable, which can win the patients' praise, but the cost of the method is high, so patients should be considered in the selection.

Objective    To evaluate the early and mid-term safety of transcatheter aortic valve replacement via transfemoral (TF), transapical (TAp) and transsubclavian (TSc) approaches by meta-analysis. Methods    We systematically searched the clinical comparative trials published from inception to June 2019 from PubMed, Web of Science, EMbase and The Cochrane Library, to evaluate the safety of transcatheter aortic valve replacement through TF, TAp or TSc approaches. The information of all-cause mortality at 30 days, 1 year, 2 years and the incidence of common complications at 30 days after operation (including pacemaker-dependent block, major vascular complications, severe bleeding events, acute renal injury and stroke) were exacted, and a meta-analysis was conducted by RevMan 5.3 software. Results    This study included 11 literatures, with a total of 7 833 patients, among whom 5 348 patients were treated by TF TAVR, 1 796 patients by TAp TAVR and 689 patients by TSc TAVR. The results of the meta-analysis were as follows: (1)  at 30 days after operation, the mortality of TF and TSc approaches were lower than that of the TAp approach (TF vs. TAp:OR=0.57, 95%CI 0.39-0.84, P=0.004; TSc vs. TAp: OR=4.12, 95%CI 1.93-8.79, P=0.000 3). There was no statistical difference between the TF and TSc approaches (TF vs. TSc: OR=0.98, 95%CI 0.38-2.51, P=0.97); at 1 year, there was no statistical difference in mortality among the three approaches (P>0.05); at 2 years, there was no statistical difference between TSc and TF or TAp approaches (TF vs. TSc: OR=1.21, 95%CI 0.95-1.54, P=0.13; TSc vs. TAp: OR=1.02, 95%CI 0.76-1.36, P=0.91). (2) The incidence of acute kidney injury after TF approach was lower than that of the TAp approach (OR=0.30, 95%CI 0.22-0.41, P<0.000 01). (3) There was no statistical difference in major vascular complications between TSc and TF or TAp approaches (TF vs. TSc: OR=0.75, 95%CI 0.38-1.49, P=0.41; TSc vs. TAp: OR=1.37, 95%CI 0.56-3.32, P=0.49). (4) There was no statistical difference in severe bleeding events between TF and TSc (OR=0.97, 95%CI 0.53-1.76, P=0.92). (5) There was no statistical difference in the incidence of postoperative stroke, pacemaker dependent block among the three approaches (P>0.05). Conclusion    TAp and TSc approaches are safe and effective. They are not only an alternative to TF approach, but also the first choice in some patients with poor condition of iliofemoral artery.

OBJECTIVETo investigate the expression of NOB1 in colorectal cancer and its relationship with the clinicopathological characteristics.

METHODSThe expression of NOB1 was detected immunohistochemically in 60 primary colorectal cancer tissues and the corresponding normal epithelia (3.0 cm away from the cancer margin) and graded according to the staining intensity and the percentage of positively stained tumor cells.

RESULTSNOB1 overexpression was found in 32 of the 60 cases (53.3%). NOB1 overexpression in the adjacent non-neoplastic tissues was found in 10 of the cases (16.7%), a rate significantly lower than that in the cancer tissues (P<0.05). NOB1 expression was not correlated to such tumor characteristics as gender, age, histological differentiation grade, depth of invasion and lymph node metastasis (P>0.05).

CONCLUSIONSNOB1 expression is higher in colorectal cancer than in normal colorectal tissues, suggesting its involvement in the tumorigenesis and progression of colorectal cancer.

Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; metabolism ; RNA-Binding Proteins ; genetics ; metabolism

An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus. The sample was incubated in 70% ethanol in a water bath for 2 h, and the extract was analyzed by HPLC using Kromasil C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution. The flow rate was 1.0 mL·min^-1. The temperature of column was 30 ℃. The detection wavelength was 280 nm. HPLC fingerprint of characteristic components of Leonurus japonicus was established. There were 12 common peaks among 25 batches of samples, and 5 of them were identified and determined. Syringic acid, leonurine hydrochloride, rutin, hyperoside or isoquercitrin showed a good linearity in the ranges of 0.426 1-85.22 ng (r = 0.999 9), 7.948-1 590 ng (r = 0.999 3), 10.20-2 040 ng (r = 1.000 0), 2.018-403.6 ng (r = 0.999 9), or 8.704-1 741 ng (r = 0.999 9), respectively. The average recoveries were 99.0%, 97.6%, 97.4%, 96.9% and 98.5% with RSD of 1.1%, 1.8%, 1.4%, 1.5% and 1.3%, respectively. The HPLC characteristic fingerprint of Leonurus japonicus was specific, and this method can simultaneously determine the content of 5 components.

Humans ; Carcinoma, Small Cell/pathology* ; Urinary Tract/pathology* ; Carcinoma, Neuroendocrine/pathology* ; Kidney Neoplasms/pathology* ; Carcinoma, Transitional Cell/pathology* ; Ureter/pathology*

The root of Scutellaria baicalensis Georgi, also called Huangqin, is frequently used in traditional Chinese medicine. In ancient China, S. baicalensis root was used to clear heat, protect the fetus, and avoid a miscarriage for thousands of years. In modern times, pregnancy-related diseases can seriously affect maternal and fetal health, but few systematic studies have explored the mechanisms and potential targets of S. baicalensis root in the treatment of pregnancy-related diseases. Flavonoids (baicalein, wogonin and oroxylin A) and flavonoid glycosides (baicalin and wogonoside) are the main chemical components in the root of S. baicalensis. This study presents the current understanding of the major chemical components in the root of S. baicalensis, focusing on their traditional uses, potential therapeutic effects and ethnopharmacological relevance to pregnancy-related disorders. The mechanisms, potential targets and experimental models of S. baicalensis root for ameliorating pregnancy-related diseases, such as recurrent spontaneous abortion, preeclampsia, preterm birth, fetal growth restriction and gestational diabetes mellitus, are highlighted.
Infant, Newborn ; Humans ; Pregnancy ; Female ; Scutellaria baicalensis ; Premature Birth/drug therapy* ; Flavonoids ; Plant Extracts/pharmacology* ; Medicine, Chinese Traditional ; Ethnopharmacology ; China

【Objective】 To evaluate the screening efficacy and practical value of the portable microfluidic biochemical analyzer in the detection of blood donors before blood donation. 【Methods】 Blood donor samples, clinical blood samples and constant quality control products were collected. Referring to the documents of ISO15189 and National Health Industry Standard, the precision and accuracy of hemoglobin (Hb) and alanine aminotransferase (ALT) were verified and compared with other detection systems. 【Results】 The MS200 biochemistry instrument has an intra-batch precision of 1.40% to 1.46%, inter-batch precision of 1.91% to 1.94%, and correctness bias of -0.9% to -1.3% for Hb test, and an intra-batch precision of 3.77% to 4.86%, inter-batch precision of 4.92% to 6.02%, and correctness bias of -3.0% to -4.8% for ALT test, which were within the range of quality requirements of industry standard. Comparison of Hb test results between MS200 biochemistry and Hb201 analyser on 1 189 peripheral blood samples from donors showed no statistically significant difference (P>0.05). 65 samples showed positive correlation between MS200 biochemistry and XS-900i automated hematology analyzer on Hb test results (R²=0.986, P=0.000). Correlation analysis of all the results of ALT detection by MS200 biochemical analyzer and AU480 biochemical analyzer in 1 065 samples showed a positive correlation (R²=0.965, P=0.000). The elevated ALT samples did not affect the Hb test results, and the samples with abnormal Hb value did not affect the ALT test results, with no interference between the two items in the detection. 【Conclusion】 The MS200 biochemical analyzer based on microfluidic technology has reliable methodological performance and can meet the need of pre-donation testing.