ObjectiveTo evaluate the efficacy of XiaochengqiMixture (XM) on promoting healing of colonic stoma. MethodsForty Wistar rats were divided into two groups randomly after colonectomy: experimental group (n=20) and control group (n=20). In early postoperatively stage rats were given gastric administration of XM in the experimental group and pure water in the control group. On day 3, 7, and 14 after establishment of animal models, laparotomy was performed in two groups of rats, respectively. Anastomotic stoma and surrounding tissues were harvested to detect the context of hydroxyproline and collagen fiber proportion by Masson dying. ResultsOn day 3 after establishment of animal models, hyperplastic collagen with small fiber was observed while no fasciculus was found. Hydroxyproline context and collagen fiber proportion of rats were higher in experimental group than those in control group (Plt;0.05). On day 7 after operation, many fasciculuses were found in two groups of rats, hydroxyproline context and collagen fiber proportion of rats were higher in experimental group than those in control group (Plt;0.01). On day 14 after operation, fasciculuses became bigger and more regular in arrangement, but there was no significant difference between the two groups (Pgt;0.05). ConclusionXM is capable of promoting healing of colonic stoma and might prevent the occurrence of anastomotic fistula.
Objective To study the migration of Schwann cells from the nerve autograft in the acellular nerve allograft of the rats in vivo. Mehtods The sciatic nerves (20 mm long) of the SD rats were harvested and prepared for the acellular nerve grafts by the chemical extraction. Then, they were observed by the gross view, HE staining, and Antilamininstaining, respectively. Another 32 female SD rats weighing 250-300 g were obtained for the study. A 2-mm-long nerve autograft was interposed between the two 10-mm-long nerve allografts to form a 22-mm-long composite. Then, the composite was placed in the muscle space, together with a sole 22-mm-long nerve allograftas a control. They were harvested at 5，10，15 and 20 days, respectively, and were then given the HE staining and the S-100 staining. Results The acellular nerve graft was semitransparent under the gross view. HE staining showed that no cell was observed within the nerve graft. Anti-laminin staining showed that the basal membrane was partially interrupted, with a positive result (dark brown). All the nerve grafts in both the groups exhibited the existenceof the cells. The S-100 positive cells were observed from the 15th day at the far ends of the two allografts of the composite; however, there were no suchcells observed within the sole nerve allograft. Conclusion Schwann cells from the sciatic nerves (2 mm- long) of the rats can migrate in the acellular nerve allograft to the far ends of the neighboring 10-mm-long nerve allografts at 15 days after operation, which offers the theoretical basis forthe repair of the longrange nerve defect by the composite of the acellular nerve allografts with the interposed nerve autograft.
ObjectiveTo study the differential expression of minichromosome maintenance protein (MCM) gene family in hepatocellular carcinoma (HCC) and to explore its survival predictive value.MethodsTranscriptome data, clinical data, and survival information of patients with HCC were extracted from The Cancer Genome Atlas (TCGA), and the differential expression of MCM gene was analyzed. The prognostic value of differentially expressed of MCM gene was studied by Cox proportional hazards regression model, the prognostic model and risk score system were constructed. On the basis of risk score, a number of indicators were included to construct a nomogram to predict the3- and 5-year survival probability of HCC patients, and to verify and evaluate their predictive ability and accuracy.ResultsThe expressions of MCM2, MCM3, MCM4, MCM5, MCM6, MCM7, MCM8, and MCM10 in HCC tissues were higher than those of normal liver tissues (P<0.05), and univariate analysis showed that they were all related to prognosis (P<0.05). Multivariate analysis showed that MCM6 and MCM10 were independent factors affecting survival of HCC patients (P<0.05). Through multivariate analysis, a prognostic model consisting of MCM6, MCM8, and MCM10 was constructed, and a risk scoring system was established. It had been verified that this risk score was an independent risk factor affecting the prognosis of patients with HCC, and the prognosis of patients with high scores were worse than those of patients with low scores (P<0.001). We used TNM stage, T stage, and risk score to construct a nomogram with a consistency index (C index) of 0.723 and draw a time-dependent receiver operating characteristic curve, the results showed that area under the curve of 3- and 5-year were 0.731 and 0.704, respectively.ConclusionsMCM6,MCM8, and MCM10 in the MCM gene family have important prognostic value in HCC. The nomogram constructed in this study can better predict the survival probability of HCC patients.
Objective To investigate the difference of minichromosome maintenance protein 2 (MCM2) mRNA expression among colonic normal mucosa, colonic adenoma and carcinoma. Methods The expressions of MCM2 mRNA were determined by real-time RT-PCR in 12 colonic normal mucosa, 33 colonic adenomas and 12 colonic carcinomas. Data were evaluated by relative expression software tool (REST-XL). Results The expressions of MCM2 mRNA elevated in turn by colonic normal mucosa, adenoma and carcinoma. The expression of MCM2 mRNA in colonic carcinomas was significantly higher than that in adenomas， as well as in normal mucosa (P=0.001), while the difference of MCM2 mRNA expressions between adenoma and normal mucosa was insignificant (P=0.184). Conclusion The difference of MCM2 mRNA expressions between adenoma and carcinoma indicated the potential value on early diagnosis for colonic carcinoma.
Six New Zealand white rabbits were under the pentobarbital anesthesia, the sciatic nerve trunk were exposed and divided. Afterwards, both the proximal and distal ends of the divided nerve trunk were incubated with Blue-SAb in the micropipe for 30 minutes at room temperature. The sensory fascieuli which were bly Blue-immunostained among unstained fasciculi and other tissues had been seen under the magnifying operating microscope (×4). The neural cells of the spinal cord and the ganglion were cultured on RPMI 1640 medium containing Bright- Blue.The growth and the metabolism of the neural cells were tested by MTT method. The t-test was used to determine the statistical difference blue staining of the neural cells was considered to be of no statistical significance (Pgt;0.05).
Objective To investigate the feature and regularity of the collagen change in bone healing during bone lengthening. Methods Bone lengthening model was made in the middle segment of the rabbit tibia. Five days after the model was established, the bone was lengthened 1.5 mm perday for 14 days. The rabbits were put to death after elongation, 7,14,21,30,40,50,60 and 70 days after elongation. The distracted area of the bone was imbedded with paraffin. After being stained by the picric acidsirius red staining, the slice was observed under polarized microscope. Results The features of the collagen change in the distracted bone were as follows: ① In the fibrous tissue of the distracted area during lengthening period and the early stage after lengthening, there was not only collagen Ⅲ but alsomuch collagen Ⅰ. ② Collagen Ⅰ, Ⅱ and Ⅲ were observed in the cartilage. ③ Collagen Ⅰ, Ⅱ and Ⅲ were also observed in the pseudogrowth plate. ④ Collagen Ⅰ took the dominance during lengtheningperiod and the late stage after lengthening. Conclusion New bone formation in bone lengthening is under the distracted force, so the collagen changes have different features compared with that in fracture healing. Collagen Ⅰ, Ⅱ and Ⅲcan be identified by picric-acid-sirius red staining and polarized microscope, so a new method for studying the collagen typing in bone repairing is provided.
Abstract： Objective To investigate the expression and correlation of phosphatase and tensin homologue deleted on chromosome ten(PTEN)， epidermal growth factor receptor(EGFR) and Ki-67 in human thymic tumors, and their possible role in tumor genesis， infiltration and metastasis. Methods The expression of PTEN， EGFR and Ki-67 were detected by using SP immunohistochemical technique in 45 cases of thymic tumors and 5 cases of normal thymic tissues. Results In 5 cases of normal thymic tissues, the expression of PTEN was bly positive, whereas EGFR and Ki -67 were weakly positive or negative. In 45 cases of thymic tumors, the positive ratio of PTEN were significantly reduced from benign thymoma， invasive thymoma to thymic carcinoma (χ2=7.808, P=0.020), but the positive ratio of EGFR and Ki-67 were gradually increased(χ2=8.032, 0.018，7.006；P=0.030). The positive ratio of PTEN， EGFR and Ki-67 protein were significantly related to Levine classification, Masaoka staging and lymph node metastasis (Plt;0.05). PTEN positive cases were negatively correlated with EGFR and Ki-67(r=－0.632，－0.653；Plt;0.01), EGFR positive cases were positively correlated with Ki-67 in thymic tumors(r=0.807,Plt;0.01). Conclusions Reduced or absent PTEN and increased EGFR and Ki-67 expression might play an important role in the genesis， invasiveness and metastasis of thymic tumors. The expression of PTEN is bly associated with the expression of abnormal EGFR and Ki-67. Detection of the three protein expressions simultaneously might be more helpful in making an early diagnosis of the tumors jndgement of theirs malignant degree，invasiveness and metastasis capacity, as well as the prognosis.
Objective To explore the appl ication of 3D nerve visual ization system in processing 2D imageinformation of human ulnar nerve acquired by series freezing tissue section, staining and scanning. And to draw the 3Danatomical atlas of human ulnar nerve through 3D Nerve visual ization software system. Methods One left ulnar nerve (frommedial fasciculus of brachial plexus to transverse carpal l igament, about 50 cm ) was taken from a fresh donated cadaver. After marked with human hair and embedded in OCT, series freezing tissue sections were made and stained with acetylchol inesterasehistochemically. Series 2D image information was obtained through high resolution scanner. Then the microstructure of ulnar nerve was reconstructed with 3D Nerve visual ization software system. Results Different cross sections of ulnar nerve have different numbers, positions and characters of the internal nerve fibers. The microstructure of ulnar nerve could be observed in magnifying visual field at any cross section after reconstructed in 3D Nerve visual ization soft ware system, which made it possible to track stereo courser of fascicles. Conclusion Reconstructed 3D Nerve visual ization software system shows the whole microstructure of ulnar nerve and the 3D stereo-structure of its internal fascicles, thus provides exact topography atlas for medical teaching and facil itates precise repair of ulnar nerve injury to improve theraputic effect.
Objective To evaluate the urine cytology silver staining combined with ultrasonography（USG）in the detection of bladder transitional cell carcinoma （TCC） recurrence after transurethral resection of bladder tumor（TURBT）in terms of sensitivity and specificity. Methods Cystoscopy was used as “gold standard”. Urine cytology combined with USG or cystoscopy was measured separately and blindly. AgNORs protein stained by silver were used in cytology with Kappa of inter-observers 0.81. For the USG, the patients were scanned with trans-rectal probe with Kappa of inter-observers 0.76. The results of urine cytology combined with USG （Positive when urine cytology and/or USG positive. Negative when both urine cytology and USG negative） were compared with “gold standard”. Results The 148 consecutive superficial TCC patients with TURBT one year previously were included in this study. Fifty seven recurrenced cases were detected. Recurrence rate was 38.51%. The sensitivity and specificity of urine cytology silver stain were 89.47% （95% CI 0.82 to 0.98） and 87.91% （95% CI 0.81 to 0.95）. Area under ROC curve was 82.22%. The sensitivity and specificity of USG were 57.90% （95% CI 0.45 to 0.71 ） and 90. 11% （ 95% CI 0.84 to 0.96）. Area under ROC curve was 73.13% . The sensitivity was improved to 94. 74% （95% CI 0.89 to 1.00） when cytology combined with USG. But specificity decreased to 84. 62% （95% CI 0.77 to 0.92 ）. Area under ROC curve was improved to 98.28%. Conclusions Urine cytology silver stain combined with USG improves the high sensitivity for follow-up TCC patients after TURBT. The non-invasive protocol is suggested.