Objective To explore a simple, effective and stable method for the isolation and purification of Kupffer cells from rat liver, enabling further study on the structure and function of these cells in vitro. Methods After laparotomy, a catheter was inserted into the portal vein and secured with artery clamp. Then, the rat liver was perfused and digested with solution Ⅰ and solution Ⅱ containing 0.05% collagenase Ⅳ respectively. The cell suspension was centrifuged with isopycnic sedimentation in a two-step Percoll gradient to harvest Kupffer cells. The isolated Kupffer cells were purified by selective adherence after 30 min of cultivation, and identified by evaluation of phagocytosis of India ink and peroxidase staining with DAB through light and electron microscopy. Results It was verified that the viability of isolated Kupfffer cells was more than 90% through Trypan blue staining. Those Kupffer cells could attach to plastic quickly and phagocytose ink, and had the appearance of “fried eggs” in positive peroxidase staining with a purity of 95%. Under the light microscopy, the appearance of newly isolated Kupffer cells was round with uniform shape and size. After two days of culture, Kupffer cells appeared to distend with irregular or stellate shape. The typical features were observed in the transmission electron micrographs. There were numerous pseudopods and occasional cup-like indentations in the cell membrane of Kupffer cells. The cytoplasm contained numerous types of lysosomes and other phagocytotic vesicles. Conclusion The method for isolating and culturing Kupffer cells in this study is effective and stable, and the biological characters are preserved in the cultured cells.
Abstract： Objective To analyze the influence of preoperative left atrial dimension （LAD） on the effectiveness of surgical radiofrequency ablation for the treatment of atrial fibrillation （AF） through a 5-year postoperative follow-up of AF patients after surgical radiofrequency ablation. Methods Clinical data of 433 patients with persistent or permanent AF who received bipolar radiofrequency ablation procedures during concomitant cardiac surgery in Beijing Anzhen Hospital from 2006 to 2009 were retrospectively analyzed. All the patients were divided into 4 groups according to their preoperative LAD：Group A， 75 patients with their LAD＜50 mm， including 22 males and 53 females with their average age of 56.50±10.05 years；Group B， 89 patients with their LAD ranging from 50 to 60 mm， including 32 males and 57 females with their average age of 55.63±10.28 years；Group C， 117 patients with their LAD ranging from 60 to 70 mm， including 41 males and 76 females with their average age of 55.13±10.96 years；and Group D， 152 patients with their LAD＞70 mm， including 68 males and 84 females with their average age of 53.22±11.49 years. Postoperative ECG records right after surgery， before discharge， at 6 months and 1，2，3，4 and 5 years during follow-up were collected. The relationship between preoperative LAD and postoperative sinus rhythm restoration rate was analyzed. Results There was statistical difference in sinus rhythm restoration rate right after surgery（P=0. 011）， before discharge（P=0. 002）， at 6 months（P＜ 0. 001） and 1 year （P＜0. 001）， 2 years（P＜0. 001）， 3 years（P＜0. 001）， 4 years（P＜0. 001） and 5 years（P= 0. 006） during follow-up among the 4 groups. Postoperative sinus rhythm restoration rates right at 6 months and 1，2，3， 4 and 5 years during follow-up was 90.4%， 89.9%， 90.3%， 91.3%， 89.1%， and 90.9% in Group A， 80.2%，79.0%，78.1%， 76.1%，72.5%，70.0% in Group B，74.7%，74.0%，71.2%，72.4%，70.0%， and 64.7% in Group C， and 61.8%，57.6%，56.8%，53.9%，50.7%，and 48.6% in Group D， respectively. Conclusion Patients with a larger preoperative LAD have a lower postoperative sinus rhythm restoration rate after surgical radiofrequency ablation for the treatment of AF.
Objective To compare the long-term outcome between breast-conserving therapy with mastectomy therapy for early stage invasive breast cancer through a Meta analysis of the randomized controlled trials published worldwide. Methods Cochrane systematic evaluation was used to search through Cochrane libraries of clinical comparative trials, PubMed, Embase, Cancer Lit, and so on. The quality of literatures was independently evaluated and cross-checked by two evaluators, indicators for assessment including death number at the end of follow-up, locoregional and total recurrence. The results were analyzed with RevMan 4.2.2 software. Results Six articles were involved in the Meta analysis with total 3 933 patients. No statistical difference was found in the death at the end of follow-up between breast-conserving therapy group and mastectomy therapy group (OR=1.05, 95% CI=0.93—1.19， P=0.45). Locoregional and total recurrence rate of breast-conserving therapy group were statistical higher than those in mastectomy therapy group (OR=1.64, 95% CI: 1.10—2.44， P=0.01； OR=1.42, 95% CI: 1.22—1.64， Plt;0.01). Conclusions Breast-conserving therapy and mastectomy therapy have comparable effects on mortality in patient with early stage invasive breast cancer, even after long-term follow up. However, breast-conserving therapy is associated with significantly greater risk of locoregional recurrence.
Objective To investigate the clinical characteristics and risk factors associated with death in critically ill patients with severe community-acquired pneumonia( CAP) .Methods A retrospective analysis was carried out in 125 hospitalized patients with severe CAP admitted from July 2008 to February 2012. Earlier clinical features were compared between 109 survival patients and 16 dead patients, and logistic regression analysis was conducted. Results The death group had more underlying diseases than the survival group( P lt;0. 05) . The heart rate at admission in the death group was significantly higher than that in the survival group( P lt;0.05) . The ratio and number of complication, the highest temperature before admission, the platelet count, the arterial blood pH, PaO2 and PaO2 /FiO2 in the death group were significantly lower than those in the survival group( P lt; 0. 05) . Logistic regression analysis showed that the number of underlying diseases, heart rate and PaO2 were predictors of death in the patients with severe pneumonia. Conclusion The number of underlying diseases, heart rate and PaO2 has highly predictive value of death for severe CAP.
Objective
To summarize the nutritional management strategies of patients undergoing electively gastrointestinal surgery.
Methods
This article reviewed the recent researches on perioperative nutritional management in electively gastrointestinal surgery, including four major directions: preoperative nutritional evaluation, glucose level control, nutritional type, and immunonutrition.
Results
At present, preoperative nutritional evaluation methods included anthropometry, laboratory tests, subjective global assessment (SGA), nutritional risk screening (NRS) 2002, Reilly nutritional risk screening, nutritional risk indicator (NRI), and so on. For preoperative nutritional assessment system, however, current data could not single out superiority for any nutritional assessment methods in the ability to predict surgery-related complications. The usage of enhanced recovery after surgery (ERAS) protocol to reduce surgical stress and preclude postoperative insulin resistance had recently been clearly linked to reductions in postoperative morbidity and adverse outcomes. There were specific criterias for perioperative parenteral and enteral nutrition in undernourished patients, who were defined in clinical guidelines recently, such as the Guidelines for Adult Perioperative Nutrition Support issued by Chinese Society of Parenteral and Enteral Nutrition (CSPEN). Several systematic reviews showed that immunonutrition could reduce both morbidity and length of stay after major electively gastrointestinal surgery.
Conclusion
Perioperative nutritional management can ensure patients benefit from nutritional support by nutritional assessment, can reduce the nutritional risk and metabolic disorder caused by operation, can achieve the goal of optimal nutrition support in surgical patients, and can ultimately reduce postoperative complications.
Objective To evaluate if performing thyroidectomy through small incision has any notable aesthetic impact on patients compared with larger incision. Methods Thirty consecutive patients underwent thyroidectomy were enrolled from March 2008 to June 2008 in this prospective randomized pilot study. The incision length was 6 cm in the small incision group and 9 cm in the larger incision group. After 3 years follow-up，the scar aesthetics were evaluated by patients and surgeons using the Patient and Observer Scar Assessment Scale （POSAS），Vancouver Scar Scale （VSS），respectively. Digital photographs about scars were taken and assessed by non-research related viewers. Results There were 13 cases who received scar aesthetic assessment in both groups. The demographic characteristics of both groups were comparative. The overall patients’ satisfactions for the small incision group and the larger incision group were （2.5±1.9） scores and （2.2±1.5）scores， respectively （P=0.55）. There were no significant differences in scar assessment scale score as for other scar assessment scales （including VSS score，PSAS score，and OSAS score） between the two groups. The evaluation of digital photographs about scars by non-research related viewers was no significant difference （P＞0.05）. Conclusion Larger cervical scar in thyroidectomy does not decrease patients’ satisfaction with their scar results.
ObjectiveTo compare the indirect calorimetry (IC) measured resting energy expenditure (MREE) with adjusted Harris-Benedict formula calculating resting energy expenditure (CREE) in the mechanically ventilated surgical critically ill patients and to evaluate the relationship between the resting energy expenditure (REE) with the severity of illness. MethodsTwenty-one patients undergonging mechanical ventilation for critical illness in the intensive care unit of general surgery between August 2008 and February 2010 were included in this study. Data during the study period of nutrition support were collected for computation of the severity of critical illness by acute physiology and chronic health evaluation Ⅱ scores (APACHE Ⅱ scores) and organ dysfunction scores (Marshall scores). MREE was measured by using IC of the MedGraphics CCM/D System within the first 7 d after nutrition therapy. CREE was calculated by using the HarrisBenedict formula adjusted with correction factors for illness at the same time. According to APACHE Ⅱ scores on admission, the enrolled patients were divided into two groups: APACHEⅡ score ≥20 scores group (n=8) and APACHE Ⅱ score lt;20 scores group (n=13), and the differences between MREE and CREE of patients in two groups were determined. ResultsThe reduction of variation tendency in CREE other than MREE in the enrolled patients within the first week of nutritional support was statistical significance (Plt;0.001). The CREE of patients 〔(1 984.49±461.83) kcal/d〕 was significantly higher than the MREE 〔(1 563.88±496.93) kcal/d〕 during the first week of nutritional support (Plt;0.001). The MREE on the 0, 1, 2, and 4 d after nutrition therapy were statistically significant lower than CREE at the same time interval in these patients (Plt;0.01), and the differences at the other time points were not significant (Pgt;0.05). There was a trend towards a reduction in APACHE Ⅱ and Marshall scores within the first week of nutrition therapy that reached statistical significance (Plt;0.001). During the first week of nutrition therapy, APACHEⅡ and Marshall scores of patients in ≥20 scores group were significantly higher than those in lt;20 scores group, respectively (Plt;0.05 or Plt;0.01), and the reductions of APACHE Ⅱ scores and Marshall scores were significant in patients of two groups (Plt;0.001). A significant positive correlation was found between CREE with APACHE Ⅱ scores (r=0.656, Plt;0.001) and Marshall scores (r=0.608，Plt;0.001) in patients within the first week after nutrition support. Although no statistically significant correlation was observed between MREE and APACHEⅡ scores (r=-0.045， P=0.563), a significant positive correlation was observed between MREE and Marshall scores (r=0.263, P=0.001) within the first week after nutrition therapy. There was no correlation between MREE and CREE (r=0.064, P=0.408) in patients at the same time interval. The reduction of MREE of patients in ≥20 scores group other than in lt;20 scores group was statistically significant within the first week after nutrition therapy (P=0.034). In addition, the MREE of patients in ≥20 scores group were not significantly different from those in lt;20 scores group (Pgt;0.05), and the mean CREE was not different in two groups patients within the first week of nutritional therapy 〔(1 999.55±372.73) kcal/d vs. (1 918.39±375.27) kcal/d, P=0.887〕. CREE was significantly higher than MREE of patients in ≥20 scores group within the first week except the 3 d and 5 d after nutrition therapy (Plt;0.05), while in lt;20 scores group CREE was significantly higher than MREE in patients only within the first 3 d after nutrition therapy (Plt;0.05 or Plt;0.01). MREE and CREE of patients in ≥20 scores group were not different from those in lt;20 scores group, respectively (Pgt;0.05).
ObjectiveTo explore the changes of focal adhesion kinase (FAK) in the fibrotic atrium of patients with valvular atrial fibrillation and explore its downstream signaling pathways.MethodsA total of 45 patients with mitral valve disease were included in this study and were divided into a valvular atrial fibrillation group (VAF, ≥6 months, 25 patients) and a sinus rhythm group (SR, 20 patients) based on having atrial fibrillation or not. The atrial appendage tissue was obtained during the operation , histopathological examination and Western blotting were performed. The degree of atrial fibrosis and changes in FAK and its downstream pathways in fibrotic myocardium were observed.ResultsThis study revealed a higher degree of atrial fibrosis in valvular atrial fibrillation and disordered cell arrangement. Expression of fibroblast differentiation marker alpha smooth muscle actin (α-SMA) was significantly increased in atrial fibrillation, and the expression of FAK and downstream AKT/S6K pathway proteins was up-regulated, while the other signal was observed, there was no significant change in ERK1/2 signaling pathway.ConclusionAtrial fibrosis in valvular atrial fibrillation is an important feature of atrial structural remodeling. We found overproduction of collagen fibers disrupted the continuity of atrial myocytes, leading to abnormal conduction and providing a matrix environment for the development of atrial fibrillation. The expression of focal adhesion kinase and downstream AKT/S6K signaling pathway in fibrotic myocardium may be involved in the process of atrial fibrosis, providing a basis for the study of its mechanism.
Objective To evaluate the impact of total thyroidectomy on health-related quality of life （HRQOL） in patients with nodular goiter. Methods The patients who underwent total thyroidectomy from Jan. 2009 to Dec. 2011 in our hospital were retrospectively analyzed with regard to the quality of life （total thyroidectomy group）. The patients with similar demographic features who underwent hemithyroidectomy during the same period were matched as control （hemi-thyroidectomy group）. The validated HRQOL instrument， which was the Euro quality of life-5D （EQ-5D）， was applied to measure the HRQOL. Comparison of HRQOL in patients of 2 groups was performed， meanwhile， the data of total thyroidectomy group was compared with data of normal population who were obtained from The Forth National Health Survey. Results There were 26 and 28 valid questionnaires returned for the total thyroidectomy group and hemithyroi-dectomy group respectively. The demographic features of patients in 2 groups were comparable. No significant variancecould be found between the 2 groups that there were no significant differences on the mobility， self-care， usual activities，pain/discomfort， anxiety/depression， and visual analogous scales （P＞0.05）. Furthermore， no significant differences in HRQOL were found in EQ-5D questionnaire compared with normal population derived from The Forth National Health Survey （P＞0.05）， except that there were more patients complained of moderate and severe pain/discomfort in the total thyroidectomy group 〔30.8% （8/26） vs.9.2% （16 330/177 501）， P＜0.01〕. Conclusion Total thyroidectomy appears to have little impact on the quality of life in the patients with nodular goiter.
Objective To investigate the threshold value for postoperative parathyroid hormone （PTH） that can predict the development of symptomatic hypocalcemia. Methods The patients received total thyroidectomy and all the operations performed by the same team from June 2010 to December 2011 in the thyroid and breast group of department of general surgery，Xuanwu Hospital of Capital Medical University were enrolled in this prospective study. Serum PTH and calcium were tested at 1-hour postoperatively and thereafter every day during hospitalization. Symptoms of hypocalcemia were recorded. Receiver operator characteristic curve was plotted to test the value of PTH assay to predict symptomatic hypocalcemia. Results There were 45 consecutive patients recruited in this prospective study，9 （20.0%） patients remained normocalcemic and 36 （80.0%） patients developed hypocalcemia postoperatively. There were 19 （42.2%） patients who developed symptoms associated with hypocalcemia and needed calcium supplement. The postoperative PTHs for normocalcemic （n=9），asymptomatic （n=17），and symptomatic （n=19） hypocalcemia patients groups were （40.5±23.7） ng/L，（18.3±9.1） ng/L，and （7.6±4.2） ng/L，respectively （P＜0.001）. The PTH value less than 13.4 ng/L could easily predict who were at high risk to develop symptomatic hypocalcemia with sensitivity of 94.7% and specificity of 76.9%. Conclusions This study suggests that a 1-hour postoperative PTH level at 13.4 ng/L can identify those individuals at risk for developing symptomatic hypocalcemia. Therefore，we recommend early calcium supplementation for these patients to decrease their postoperative morbidity from symptomatic hypocalcemia.
