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 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 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 study the mechanism of gastric bypass operation on treatment of type 2 diabetes mellitus, recognize the etiology and pathogenesy of the disease and frame therapy strategy for type 2 diabetes mellitus. Methods The literatures about gastric bypass operation on treatment of type 2 diabetes mellitus, including clinical cases reports and evidence-based studies were reviewed. Results Gastrointestinal bypass operation was regarded as an effective treatment for type 2 diabetes mellitus. There were three hypotheses of therapy mechanism: early delivery of nutrients to the distal intestine, exclusion of the proximal intestine and incretin/anti-incretin. Conclusion Gastrointestinal bypass operation is now considering as an effective treatment, there is still a lack of basic experimental studies to clarify the mechanism.
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.
The treatment of patients suffering from both severe coronary artery disease (CAD) and malignancy is different from those who have isolated CAD or tumor, because their body condition is poorer and the lesions of coronary artery are severer, which poses severe challenges to doctors. At present, the surgical treatment for these patients mainly includes staged operation and simultaneous surgery. Staged operation, which separates coronary artery revascularization surgery from tumor resection, has a lower incidence of postoperative complications, but tumor progression may occur during the waiting period. Simultaneous surgery combines heart procedure with tumor resection at the same time, shortens the treatment period of patients, overcomes the defect of tumor progression that may occur during the waiting period of the staged surgery, but the incidence of postoperative complications is higher. The progress of surgical treatment in patients with CAD combined with malignant tumor is summarized.
Objective To summarize the recent progress in pathogenetic, diagnostic and therapeutic researches on the intestinal barrier dysfunction (IBD) of severe acute pancreatitis (SAP). MethodsThe advancement of IBD in SAP, which was published recently at home and abroad, was collected and reviewed. Results The pathogenesis of IBD in patients with SAP was complex. Ischemia-reperfusion injury, endotoxin, inflammatory mediators and gastrointestinal hormone played an important role in the process of IBD. There were many ways to detect IBD, and the ratio of lactulose and mannitol, plasma diamine oxidase were relatively ideal markers. Medical therapies, such as treatment of SAP and maintaining the perfusion of intestines, were essential to cure IBD. On this basis, the propulsives, nutritional support and traditional Chinese drugs should be administered reasonably. Conclusions IBD is a sophisticated process of pathophysiology. In recent years, abundant of animal experiments and clinical researches have provided new clue for prevention and cure of IBD, but further researches are still needed on the mechanism of the cells and molecules implicated.
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).