The effect of Danshen extract on lipoprotein-associated phospholipase A2 levels in patients with stable angina pectoris: study protocol for a randomized controlled trial - the DOLPHIN study.

BACKGROUND: Lipoprotein-associated phospholipase A2 (Lp-PLA2), a biomarker of oxidation and inflammation, has been associated with increased coronary artery disease risk. To date, very few studies have examined the Chinese herbal drug Danshen or its extract on Lp-PLA2 in patients with stable angina pectoris. In this study, we aim to investigate the effect of Danshen extract on Lp-PLA2 level in patients with stable angina. METHODS/DESIGN: This is a randomized, single-blind, placebo-controlled, adaptive clinical trial. A total of 156 patients meeting the eligibility criteria will be randomly assigned to either the Danshen extract (DanshenDuofensuanyan injection and Danshen drop spill) group or the placebo group in a 1:1 ratio. Participants will then undergo treatment with DanshenDuofensuanyan injection or placebo (glucose) during hospitalization, followed by open-label Danshen drop spill (30 pills/day) in Danshen extract group for 60 days after discharge. Because this is an adaptive trial, two interim analyses are prospectively planned. These will be performed after one-third and two-thirds of the patients, respectively, have completed the trial. On the basis of the results of these interim analyses, a data monitoring committee will determine how to modify aspects of the study without undermining the validity and integrity of the trial. The primary outcome measure is the serum level of Lp-PLA2 in the Danshen extract group and the placebo group. The secondary outcomes include the proportion of patients who show a clinically significant change, which is defined as at least a 20-point improvement in angina frequency score on the Seattle Angina Questionnaire and the carotid intima-media thickness, which will be measured using ultrasound. Other secondary efficacy and safety outcomes will also be assessed. DISCUSSION: This study will provide evidence that Danshen extract is beneficial for stable angina and may establish a possible mechanism of Danshen treatment effects on cardiovascular disease. This study may also validate an objective blood test (LP-PLA2 level) for assessing the effectiveness of Danshen therapy in patients with stable angina pectoris. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02870764 . Registered on 13 August 2016.

Selectivity evaluation and separation of human immunoglobulin G, Fab and Fc fragments with mixed-mode resins.

Adsorption selectivity is critical important for mixed-mode chromatography with specially-designed ligands. Human immunoglobulin G (hIgG), Fc and Fab fragments were used in the present work to evaluate adsorption behavior and binding selectivity of four mixed-mode resins with the ligands of 4-mercatoethyl-pyridine (MEP), 2-mercapto-1-methylimidazole (MMI), 5-aminobenzimidazole (ABI) and tryptophan-5-aminobenzimidazole (W-ABI), respectively. The resins showed an obvious pH-dependent adsorption behavior. High adsorption capacities were found at neutral pH for hIgG, Fc and Fab, and almost no adsorption happened under acidic conditions. An adsorption selectivity index was proposed to evaluate separation efficiency. High specificity of hIgG/Fc was found at pH 8.9 for MEP resin, and for W-ABI resin at pH 8.0 and 8.9. In addition, isothermal titration calorimetry was used to evaluate ligand-protein interactions. Finally, the separation of hIgG and Fc (1:1) was optimized with mixed-mode resins, and the best separation performance was obtained with W-ABI-based resin. Loading at pH 8.0 resulted in the flow through of Fc with purity of 90.4% and recovery of 98.8%, while elution at pH 3.6 provided hIgG with purity of 99.7% and recovery of 86.5%.

A microcalorimetric study of molecular interactions between immunoglobulin G and hydrophobic charge-induction ligand.

Hydrophobic charge-induction chromatography (HCIC) with 4-mercaptoethyl-pyridine (MEP) as the ligand is a novel technology for antibody purification. In this study, isothermal titration calorimetry (ITC) was used to evaluate the molecular interactions between MEP ligand and immunoglobulin G (IgG). Three types of IgG molecules including human IgG (hIgG), bovine IgG (bIgG) and a monoclonal antibody (mAb) were investigated with human serum albumins (HSA) and bovine serum albumin (BSA) as the comparison. The thermodynamic parameters obtained from ITC were compared with the adsorption data. The results indicated that MEP binding to protein at neutral pH was entropy driven and induced by multimodal molecular interactions that was dominated by hydrophobic forces. The interactions between MEP and IgGs were stronger than that of albumins, which resulted in high binding affinity of IgGs. Moreover, the effects of pH and salt addition on MEP-hIgG binding were studied. The change of enthalpy increased obviously with the decrease of pH, which revealed that the electrostatic forces dominated the MEP-hIgG interactions at acidic condition and caused typical charge-induced elution of HCIC. Salt addition influenced both hydrophobic and electrostatic interactions. With the increase of salt concentration, the hydrophobic interactions decreased first and then increased, while the electrostatic interactions showed the opposite trend. This resulted in trade-off between the multimodal interactions, which caused the salt-tolerant property of MEP resin. In general, ITC studies revealed the molecular mechanism of three critical characteristics of HCIC, multimodal interactions, pH-dependent and salt-tolerant properties.

[Clinical outcomes of percutaneous kyphoplasty under local anesthesia for osteoporotic vertebral compression fractures].

OBJECTIVE: To analyze the clinical outcomes of percutaneous kyphoplasty under local anesthesia for osteoporotic vertebral compression fractures. METHODS: In this study, 76 elderly patients with osteoporotic vertebral compression fractures undergoing percutaneous kyphoplasty were followed up for 2-3.1 years. They were divided into 3 groups according to different ages: 60-69 yrs (A group), 70-79 yrs (B group) and 80- 91 yrs (C group). Pre- and post-operative and last follow-up evaluations were conducted. And the parameters of bone mineral density (BMD), kyphotic angle, change of visual analog scale (VAS), Oswestry disability index, average vertebral body height, complications and costs of hospitalization were recorded and analyzed. RESULTS: BMD decreased with advancing age and had statistical significance between three groups (P < 0.05). Three thoracic spine fractures and 6 lumbar spine fractures could not be detected with digital radiography and were observed only on magnetic resonance imaging (MRI). The pre-operative levels of visual analogue scale and Oswestry disability index increased in all groups.No statistical significance existed between A and B groups. But there was statistical significance between A or B and C groups (P < 0.05). Pre- and post-operative assessments showed that statistically significant improvements were found in visual analogue scale and Oswestry disability index in all groups (P < 0.05). And statistically significant improvements were found for the pre- and post-operative kyphotic angles and vertebral body heights in A, C group and B groups (P < 0.05). The sites for symptomatic leakage of cement included paravertebral vein (n = 2), intervertebral disc (n = 1) and paravertebral space (n = 2). Adjacent vertebral fracture occurred in 1 patient at 17 months and underwent percutaneous kyphoplasty. The mean operative duration was 28 minutes per vertebrae and the mean cost of hospitalization at RMB yuan 33 778. CONCLUSION: As a simple and safe procedure for osteoporotic vertebrae compression fractures, percutaneous kyphoplasty may relieve pain quickly, restore vertebral height, prevent further fractures and improve the patient's quality-of-life.

Caprylate as the albumin-selective modifier to improve IgG purification with hydrophobic charge-induction chromatography.

Hydrophobic charge-induction chromatography (HCIC) is a novel downstream bioprocessing technology for antibody purification and it has several advantages over traditional purification processes. However, its separation selectivity still needs to be improved. In this work, sodium caprylate (NaCA) was used as the selective modifier to improve IgG purification from serum albumin containing feedstock with a typical HCIC resin, MEP HyperCel. The effects of NaCA on the adsorption equilibrium of bovine serum immunoglobulin G (IgG) and bovine serum albumin (BSA), as well as the dynamic binding and displacement behaviors were investigated. The binding and elution behaviors of these two proteins in the column were studied. It was found that adding 50-75 mM NaCA in the liquid phase could effectively reduce the adsorption of BSA on the MEP resin, but the same treatment has little influence on the adsorption of IgG. Moreover, the mechanism of the competitive binding between caprylate and MEP ligands on the surface of BSA is discussed. It was found that by controlling NaCA addition in the loading or washing buffer, the process efficiency of IgG purification from BSA containing feedstock can be improved, and the purity of IgG could reach to over 98%. The results indicated that caprylate could be a promising albumin-selective modifier to improve the separation efficiency of antibodies with the HCIC process.

[Locked plating with minimally invasive percutaneous plate osteosynthesis versus intramedullary nailing of distal extra-articular tibial fracture: a retrospective study].

OBJECTIVE: To compare the efficacies of minimally invasive plate osteosynthesis (MIPPO) and interlocking intramedullary nailing (IMN) in the treatment of extra-articular fractures of distal tibia. METHODS: Retrospective reviews were conducted for 126 patients with extra-articular distal tibia fractures. Treatment was either MIPPO (n = 61) or IMN (n = 65). The outcomes were assessed by comparing operating duration, time to union, the last follow-up American Orthopedic Foot and Ankle Society (AOFAS) score and complication rate. RESULTS: The average follow-up period was 23.7 (12-53) months. In the minimally invasive plate osteosynthesis group, there were deep infections (n = 2), superficial infections (n = 5), delayed union (n = 2), malunion (n = 2) and knee joint pain (n = 10) were observed. In addition, the average operating duration (85.9 ± 18.9 min), average time to union (17.3 ± 3.8 weeks) and average AOFAS (83.2 ± 11.9) were analyzed. In the interlocking intramedullary nailing group, there were delayed union (n = 3), malunion (n = 12) and knee joint pain (n = 22). And the average operating duration (83.3 ± 15.7 min), average time to union (16.5 ± 3.1 weeks) and average AOFAS (84.9 ± 12.0) were analyzed. No statistical significance existed in operating duration, time to union and the last follow-up AOFAS between two groups (P > 0.05). However, the rates of malformation and knee joint pain were higher in the intramedullary nail group than those in the plate group. And the difference was statistically significant (P = 0.015, P = 0.025). CONCLUSION: Both MIPPO and IMN are effective for extra-articular fractures of distal tibia. However, the former has the advantage of lowers rate of malformation and knee joint pain. Therefore a surgeon should consider the degree of injury while managing extra-articular fracture of distal tibia.

[Prognostic effect of postoperative resection-margin status for intraoperative positive resection margins in patients with advanced gastric cancer].

OBJECTIVES: To investigate prognostic effect of postoperative resection-margin status for intraoperatively positive resection margin in advanced gastric cancer and discuss the treatment choice for intraoperatively positive resection margins. METHODS: A retrospective study was investigated in 64 advanced gastric cancer patients with positive resection margin after potentially curative resection. The survival between 50 patients who was re-excised to a negative resection margin (NR group) and 14 patients who were left with positive resection margin (PR group) was compared. Prognostic factors were analyzed using univariate and multivariate Cox regression model analysis. RESULTS: The median survival in the PR group was 17.0 months (95%CI: 11.6 - 22.4) as compared with 23.0 months (95%CI: 20.5 - 25.5) in the NR group (P = 0.045). However, resection-margin status lost significance on multivariate analysis. In the subgroup of D2 lymphadenectomy, the median survival in the PR group and NR group were 17.0 months (95%CI: 12.0 - 22.0) and 24.0 months (95%CI: 19.8 - 28.1) respectively; multivariate analysis further identified resection margin status as an independent prognostic factor. CONCLUSIONS: Re-excision for intraoperatively positive margin to negative margin improves the prognosis of the patients with advanced gastric cancer, and re-excision is the first choice when intraoperative frozen section detects a positive margin. Routine frozen section of resection margin should be mandatory in all advanced gastric cancer undergoing potentially curative surgery.

Enhancing IgG purification from serum albumin containing feedstock with hydrophobic charge-induction chromatography.

Hydrophobic charge-induction chromatography (HCIC) with 4-mercaptoethyl-pyridine (MEP) as the ligand is a novel technology for antibody purification, however, the separation selectivity still needs to be improved for the applications, especially for the impurity of serum albumin. In this study, with bovine serum immunoglobulin G (IgG) as the model, the purification of IgG from the serum albumin containing feedstock was developed with the commercial HCIC resin MEP HyperCel, focusing on the optimization of operation pH and salt addition. The adsorption isotherms of IgG and bovine serum albumin (BSA) were investigated at different pHs, and the binding and elution behaviors of two proteins in the column were also studied at varying pHs. In addition, the protein-ligand interactions were investigated with some additives in the buffer. It was found that the conditions of pH 6 with 0.1 M NaCl or pH 8 could be used to effectively remove BSA from the MEP resin without the influence on IgG adsorption. Two modes with control of loading or washing buffer were tested to enhance the purification of IgG from BSA containing feedstock, and the purity of IgG was improved to about 95% compared with 62.9% for the control. The results demonstrated that the control of loading pH or the addition of NaCl in the buffer might be an effective method to improve the purification of antibody with the HCIC process.

Clinicopathologic variables predicting tumor response to neoadjuvant chemotherapy in patients with locally advanced gastric cancer.

BACKGROUND AND OBJECTIVES: To identify clinicopathologic variables that could predict pathologic tumor response to neoadjuvant chemotherapy for patients with locally advanced gastric cancer. METHODS: The study enrolled 108 patients who underwent neoadjuvant chemotherapy followed by surgery between July 2004 and December 2010. Tumor responses to neoadjuvant chemotherapy were assessed in terms of tumor regression. Statistical analyses were performed to identify factors associated with pathologic tumor response. RESULTS: Tumor regression was found in 22.2% (24/108) patients, patients with tumor regression observed better overall survival as compared to that of patients without tumor regression. Univariate and multivariate analyses observed that both tumor differentiation and tumor size were independent predictors of tumor regression. CONCLUSIONS: This study suggests that both tumor differentiation and tumor size is the most important clinical predicator of pathologic tumor response, it may be of benefit in the selection of treatment options in locally advanced gastric cancer.

Neoadjuvant chemotherapy versus surgery alone for locally advanced gastric cancer: a retrospective comparative study.

BACKGROUND/AIMS: Preoperative chemotherapy is considered an effective treatment option for patients with gastric cancer. We retrospectively evaluated neoadjuvant chemotherapy with oxaliplatin, leucovorin and 5-flurouracil (OLF) in patients with locally advanced gastric cancer to determine its feasibility, as well as impact on the curative resection rate and patients' survival. METHODOLOGY: A total of 87 patients with locally advanced gastric cancer that underwent preoperative chemotherapy combined with surgery or surgery alone were randomly matched according to the clinical TNM stage. The clinical responses to chemotherapy were assessed. The curative rate, postoperative complications and patients' survival between both groups were compared. RESULTS: The two groups were well matched. Complete or partial response was observed in 51.7% (15/29) of patients in the OLF group, and three (10.3%) of them had complete pathologic response. The curative resection rates were 89.7% in the OLF group and 77.6% in the surgery alone group. The postoperative complications were equal for both groups. The mean survival is 20.6 months in the OLF group vs. 19.9 months in the surgery alone group (p=0.02). CONCLUSIONS: Neoadjuvant chemotherapy using OLF combination is active in gastric cancer and the toxicity level is acceptable. This treatment improves the curative resection rate and patients' survival in locally advanced gastric cancer.

Laparoscopic local resection based on sentinel node evaluation for early gastric cancer: a preliminary report.

We previously reported that lymphatic mapping using isosulfan blue can be used to identify sentinel nodes (SNs). This study was undertaken to evaluate the feasibility of using the SN technique in treating early gastric cancer and to explore its usefulness for minimal invasive surgery. Twenty-three patients with early gastric cancer who underwent SN biopsy were retrospectively evaluated. Based on SN evaluation, individualized surgery was performed in five patients with T1N0M0 gastric cancer. When pathological examination of frozen sections revealed metastasis in SNs, we performed a standard D2 gastrectomy. Laparoscopic local resection was applied when the SN biopsy was negative. Our results showed that the success rate with SN biopsy in early gastric cancer was 100%, as were the accuracy, sensitivity, and specificity. All five patients with early gastric cancer had SNs negative for metastases both by frozen section and by postoperative pathology. Thus, all these patients underwent laparoscopic local resection without extended lymphadenectomy. We conclude that SN biopsy is a useful tool to individualize the operative procedure, and laparoscopic local resection can be safely performed using SN guidance in selected patients with early gastric cancer.

Different criteria for radioactive sentinel lymph nodes has different impact on sentinel node biopsy in breast cancer patients.

BACKGROUND AND OBJECTIVES: This study set out to determine the impact of different criteria for radioactive sentinel lymph nodes (SLNs) on sentinel lymph node biopsy (SLNB), and the optimal criteria for radioactive SLNs. METHODS: Eighty-four breast cancer patients with cT1-2N0M0 were studied prospectively. Filtered technetium 99 m sulfur colloid was injected in peritumor parenchyma. Three different definitions of SLNs were adopted in each patient: (1) the lymph node with the highest radioactivity. (2) Any lymph node with an in vivo hot spot-to-background activity ratio of at least 3:1 or an ex vivo SLN-to-non-SLN ratio of at least 10:1. (3) All radioactive hot nodes. RESULTS: With three different definitions, the success rate of sentinel node biopsy were all 96.4%, the sensitivity was 78.9%, 92.1%, 97.4% respectively; false negative rate 21.1%, 7.9%, 2.6% respectively; predicting accuracy 90.1%, 96.3%, 98.8% respectively (P < 0.05). The first, the first two, the first three, and the first four highest radioactive sentinel nodes identified 81.1%, 89.2%, 94.6%, 100% of the positive-SLN patients, respectively. CONCLUSIONS: The different criteria for radioactive SLNs had different impact on the accuracy, sensitivity, and false negative rate, but not on the success rate, during sentinel node biopsy using radioactive sulfur colloid in breast cancer patients. The first four highest radioactive sentinel nodes could accurately predict the status of axillary metastases. J. Surg. Oncol. 2007;95:635-639. (c) 2007 Wiley-Liss, Inc.

[Clinical evaluation of laparoscopic radical resection of colon cancer].

OBJECTIVE: To investigate the clinical effect of laparoscopic radical resection of colon cancer. METHODS: Patients with colon cancer who underwent radical resection during January 2000 to January 2004 in Sir Run Run Shaw Hospital (Medical College of Zhejiang University) were divided into open and laparoscopic groups for a non-randomized case-control study. All the patients were followed up and their clinical and pathological results were compared. RESULTS: No difference was seen between the two groups on the patients' age, gender, lymph node numbers and Dukes staging. The laparoscopic group had a longer mean operation time [(182 +/- 62) min vs (141 +/- 37) min] and shorter hospital stay [(5.3 +/- 1.9) d vs (8.2 +/- 1.2) d] than the open group. All surgical margins were pathological negative. Post-operation follow up was 12-18 months (mean 21 months). No incision recurrence was found in neither group. As far the local recurrence and distal metastasis rate were similar in both groups. CONCLUSIONS: Laparoscopic radical resection of colon cancer has minimal invasion and shorter recuperation than open procedure, the radicalness and either local recurrence or distal metastasis rate after surgery are similar when compared with open group.