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.
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.
Objective To determine the frequencies and patterns of gastrointestinal wall thickening at muti-slice computed tomography (MSCT) in patients with hepatic cirrhosis. Methods One hundred and nine patients with cirrhosis and 130 patients without cirrhosis for gastrointestinal wall thickening were retrospectively analyzed by the abdominal MSCT scans. The frequencies of wall thickening were determined in the cirrhosis patients and in those without cirrhosis. The segmental distribution, symmetry and enhancement pattern were evaluated in all patients with cirrhosis for gastrointestinal wall thickening. Results Gastrointestinal wall thickening was seen in 72 cases (66%) with cirrhosis patients and in 12 cases (9%) without cirrhosis patients (Plt;0.005). The jejunum and ascending colon were the most common sites of gastrointestinal wall thickening, which was involved in 32 and 31 patients respectively. The scans of 46 (64%) patients with gastrointestinal wall thickening showed multisegmental distribution. Gastrointestinal wall thickening was concentric and homogeneous in all patients with cirrhosis. Conclusion Gastrointestinal wall thickening is common in patients with hepatic cirrhosis. It frequently involves multiple segments. The jejunum and ascending colon are the most common sites of involvement. MSCT plays an invaluable role in diagnostic evaluation of bowel wall thickening in patients with hepatic cirrhosis.
ObjectiveTo investigate the role of intraoperative frozen section pathology in central lymph node metastasis of papillary thyroid microcarcinoma (PTMC), and to analyze the risk factors of central lymph node metastasis.MethodsClinical data of 481 patients diagnosed with PTMC from January 2015 to June 2019 in our hospital were included. The consistency of frozen pathological results of intraoperative prelaryngeal lymph nodes, pretracheal lymph nodes, and paratracheal lymph nodes with postoperative paraffin pathological results, as well as the relationship between the numbers of intraoperative lymph nodes sent for examination and postoperative pathological results were analyzed. Then the Kappa value were calculated respectively. Furthermore, univariate and multivariate analysis were used to analyze the factors affecting central lymph node metastasis.ResultsCentral lymph node metastasis was found in 207 patients with PTMC (43.0%). Of the 207 patients, 192 patients were examined by frozen section, with 139 patients had positive results. The Kappa value of prelaryngeal lymph nodes, paratracheal lymph nodes, pretracheal lymph nodes, and central lymph nodes were 0.300, 0.643, 0.560, and 0.755, respectively (P<0.001). Simultaneous intraoperative examination of three anatomic lymph nodes in the central region has a high accuracy in evaluating whether there was lymph node metastasis. The consistency test between intraoperative frozen and postoperative paraffin pathological results showed that when the number of lymph nodes was less than 5, the Kappa value was 0.690 (P<0.001), and when more than or equal to 5, the Kappa value was 0.816 (P<0.001). The results of logistic regression showed that, maximum value of tumor diameter, tumor number, and thyroid capsule involvement were risk factors for central region lymph node metastasis in PTMC (P<0.05).ConclusionsCentral region lymph node metastasis in PTMC was common. Prelaryngeal lymph nodes, pretracheal lymph nodes, and paratracheal lymph nodes should be selected for frozen pathological examination during the operation, which could effectively indicate whether the central lymph nodes were involved. And combined with the risk factors of lymph node metastasis, such as maximum value of tumor diameter, number of tumors, and thyroid capsule involvement, a more accurate individualized operation plan can be designed for patients.
Objective To evaluate the effect of neoadjuvant chemotherapy on the expression of CXCR4 in breast cancer and its clinical significance.Methods The clinical data of 59 patients with breast cancer of stage Ⅱ and stage Ⅲ underwent neoadjuvant chemotherapy with paclitaxel plus epirubicin for 3 cycles between April 2005 and March 2009 were retrospectively analyzed. The expression of CXCR4 in the breast cancer tissues before and after neo-adjuvant chemotherapy was examined by immunohistochemistry and its relationship with clinicopathologic factors was analyzed.Results The CXCR4 positive expression was observed in 56 patients with breast cancer (94.9%), but not in corresponding nontumor normal tissues. The expression level of CXCR4 was correlated to lymph nodes metastasis (P=0.019) and breast cancer stage (P=0.040), but it was not correlated to age of patients, tumor size, grade, hormone receptor (ER and PR)， and HER2 status. The expression level of CXCR4 was significantly decreased after neoadjuvant chemotherapy. Decline extent of CXCR4 expression after chemotherapy and CXCR4 expression level were not correlated to the effect of neoadjuvant chemotherapy, while the effect of chemotherapy in patients expressed CXCR4 in cluster distribution was better than that in scattering distribution (P=0.015). Conclusion The decline extent of CXCR4 expression level after paclitaxel combined with epirubicin neoadjuvant chemotherapy is not correlated to the efficacy, but its expressing distribution may be considered as an index to the effect of neoadjuvant chemotherapy.