AIM: To investigate the effect of retinoic acid on proliferation of renal interstitial myofibroblasts in a rat model of the left unilateral ureteal obstruction (UUO). METHODS: UUO model was established by unilateral ligation of ureter in 36 SD rats. Rats were divided into 2 groups: Retinoic acid-treated group and control group, each with 18 rats. UUO rats were treated with either daily subcutaneous injection of 10 mg/kg body weight of all trans-retinoic acid or vehicle alone two days before the operation until being sacrificed. Groups of 6 rats were killed on day 3, 7 and 12 after ligation of the left ureter. The percentage of renal tubular lesion, interstitial fibrosis score, the number of interstitial myofibroblasts, the number of proliferating myofibroblasts and the expression of TGF?-1 mRNA were determined. RESULTS: There were significant accumulation and local proliferation of myofibroblasts in the interstitium of the UUO rats. On day 7 of the UUO model, the percentage of tubular lesions and interstitial fibrosis score were significantly lower in the retinoic acid-treated group than those in control group [(15.9?2.0)% vs (27.3?2.2)% and (0.47?0.12) vs (1.65?0.18), P

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To study the process of hydroxypropyl-β-cyclodextrin (HP-β-CD) inclusion techniques for osthol. Methods The inclusion complex of osthol and HP-β-CD was prepared by unsaturated water solution and freeze-drying technique. Inclusion techniques were selected by screening on quadratic orthogonal rotation combination design method, and the entrapment efficiency was identified by HPLC. Results The optimal technical conditions were as follows:the ratio of HP-β-CD and drug was 4.5∶1;temperature for electric mixer was 35 ℃;the stirring time for thermostatic waterbath was 210 min. Conclusion This method is reasonable and it may have a prosperous future of development and application.

Objective To evaluate the sensitivity, repeatability and accuracy of microarray digital PCR system in detecting JAK2 V617F mutation, which was closely related to myeloproliferative neoplasms (MPN).Methods All of the 31 MPN patients with JAK2 V617F mutation, including 18 cases of polycythemia vera(PVs),11 primary thrombocythemias (ETs) and 2 primary myelofibrosis (PMFs), were collected from Huashan Hospital, Fudan University during 2014 -2015, while 10 normal controls and 6 cases with abnormal increased hemoglobin were involved.Human erythroleukemia cell line ( HEL ) and colorectal cancer cell SW480 were used as the mutant and the wild type control, respectively.The sensitivity of microarray digital PCR were verified by detecting the gradient diluted mutation standard harboring 30%, 10%, 1%, 0.1%and 0.01%mutant allele burden, respectively .Repeatability was evaluated by detecting 1%and 10% mutated samples for 5 times, respectively.MGB probe real time PCR was selected as the reference method to verify the accuracy of the digital PCR.Results With digital PCR, the accurate quantitation of JAK2 V617F mutation was achieved down to 0.1%, which is approximate to 0.16 copies per microliter.The results obtained from the two kinds of technique showed a high correlation by linear regression analysis (R2 =0.998 3).The results of repeated samples showed CVs as 17.18% for 1%mutant allele burden and 7.50%for 10%.Among all cases, the 31 patients known mutated were detected as positive and 10 controls as negative by both digital PCR and Real time PCR.In another 6 cases, 2 were found JAK2 V617F mutation of low allele burdens of 0.37% and 0.18% by digital PCR but detected as negative by real time PCR.Conclusions Microarray digital PCR offers a higher sensitivity and better repeatability than real time PCR which could help detect rare JAK2 V617F mutations in MPNs accurately.

Objective To investigate the feasibility and primary therapeutic effect of inverted Y-shaped self-expandable metal stent for complex airway stenosis.Methods On the standpoint of the peculiar anatomic structure and the pathological changes of complex airway stenosis,we designed the inverted Y-shaped self- expandable metal stent.Under the fluoroscopic guidance,7 stents were implanted in 7 cases of airway complex stenosis.Results The inverted Y-shaped self-expandable metal stents were placed seccussfully,with instantaneous relief of dyspnea and improvement of living quality.Conclusion The placement of inverted Y- shaped self-expandable metal stent is feasible and safe for treating airway complex stenosis.(J Intervent Radiol, 2007,16:92-94)

Fatigue is a common complication of type 2 diabetes mellitus (T2DM). We examined the relationship between T2DM fatigue and the skeletal muscle 5-hydroxytryptamine (5-HT) system. In animal experiments, a T2DM model was established in mice by feeding a high-fat diet with intraperitoneal injection of streptozotocin. The mice were treated with the 5-HT_2A receptor antagonist sarpogrelate hydrochloride (SH) and the 5-HT synthesis inhibitor carbidopa (CDP) (separately and in combination). In cell culture experiments, C2C12 cells were stimulated with D-glucose, palmitic acid or 5-HT. 5-HT_2AR, 5-HT synthesis and 5-HT degradation were inhibited by SH, CDP, or monoamine oxidase A (MAO-A) inhibitor. The animal experiments were in accordance with the regulations of the Animal Ethics Committee of China Pharmaceutical University. The results showed that 5-HT_2AR, 5-HT synthase and MAO-A were expressed in mouse skeletal muscle and C2C12 cells. The expression of these proteins was significantly up-regulated in T2DM mice or when C2C12 cells were exposed to palmitic acid and D-glucose; palmitic acid was a stronger stimulant of their expression than D-glucose. Rotating rod experiments and biochemical index tests have shown that T2DM fatigue is associated with an increase in skeletal muscle 5-HT_2AR, 5-HT synthesis and 5-HT degradation. 5-HT_2AR mediates the expression of MAO-A and the synthesis of 5-HT, which indirectly regulates the degradation of 5-HT. MAO-A regulates cell inflammation, mitochondrial ROS production and membrane potential depolarization by mediating 5-HT degradation. MAO-A also inhibits the expression of peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1), carnitine palmitoyltransferase-1 (CPT1) and ATP synthase-6 (ATP6), thus inhibiting mitochondrial functions such as fatty acid β oxidation and ATP synthesis. SH and CDP can effectively treat T2DM fatigue, and can also reduce blood glucose and blood lipid, and the combination of SH and CDP has a clear synergistic effect.

OBJECTIVETo investigate the clinico-pathologic characteristics, treatment and prognosis of thyroid carcinoma in childhood and adolescents.

METHODSFrom 1984 to 1997, 86 cases with thyroid carcinoma in childhood and adolescent treated were summarized.

RESULTSAll cases underwent operation with adjuvant therapy. Pathologically, papillary carcinoma was diagnosed in 73 (84.9%), follicular carcinoma in 6 (7%), papillary-follicular carcinoma in 4 (4.7%) and medullary carcinoma in 3 (3.5%). Cervical lymph node metastasis was found in 59 cases (68.6%), 16 of which with both thyroid carcinoma and bilateral cervical lymph node metastasis (27.1%). Lung metastasis was found in 11 cases. Recurrence occurred in 6 cases after operation. Compared with the thyroid carcinoma in adult patients, cervical lymph node metastasis, bilateral involvement of the thyroid gland with bilateral cervical nodes and lung metastasis rate were more commonly seen in childhood and adolescence. All but 2 patients had been followed up for more than 5 years, 41 patients for more than 10 years. The 5-year and 10-year survival rate was 95.3% (82/86) and 87.8% (36/41), respectively.

CONCLUSIONThe clinical manifestations of childhood and adolescent thyroid cancer are generally not pathognostic which may lead to misdiagnosis. Surgery is the main method in the comprehensive treatment with a good prognosis. The therapy with (131)I after operation was beneficial for some patients accompanied with lung metastasis.

Adolescent ; Adult ; Cell Differentiation ; Child ; Child, Preschool ; Female ; Humans ; Male ; Prognosis ; Thyroid Neoplasms ; mortality ; pathology ; surgery

Objective To explore rationale and clinical application of simplified modified radical thyroideetomy for differentiated thyroid carcinoma.Methods From Jan.2007 to Jun.2010,349 cases of differentiated thyroid carcinoma received simplified operative procedure based on standard modified radical thyroidectomy.The simplified procedure took a low small collar incision(about 10-12 cm).In separating upper and lower skin flaps,subcutaneous tissues covering posterior triangle of neck and posterior edge of sternoeleidomastoid muscle were spared to protect sensory nerves.Subtotal thyroidectomy Was performed to resect the affected lobe,isthmus,and the majority of opposite lobe without considering the size of primary tumor or whether metastasis to the neck lymph nodes happened.Soft tissues of the mainly metastatic areas(Ⅱ a、Ⅲ、Ⅳ、Ⅴb)were cleared.The accessory nerve was not exposed routinely to avoid stimulation.Lymph nodes metastasis in different areas was recorded respectively.Complications in different operative modes were compared.Results Compared with standard modified radical thyroidectomy,the simplified mode had shorter scar-and no limit of neck mobility.Because of muscles and nerves pemervation,movement dysfunction and abnormal sensation of neck and shoulder decreased obviously.The operation duration was shortened.Cervical lymph node status Was evaluated,which provided basis for prognosis judgment and comprehensive treatment.Conclusions The simplified modified radical procedure has the benefit of decreased trauma while maintains the similar recurrence rate compared to modified radical thyroidectomy.It improvs the life quality of patients.This procedure fits the principle of functional radical neck dissection better.

Objective: To investigate the roles of N-acetyl-L-cysteine (NAC) against binge drinking-induced fatty liver in mice. Methods: SPF male C57BL/6 mice were randomly divided into 3 groups, i.e. control group, model group, and NAC/ethanol group (n=10). Mice in model and NAC/ethanol groups were exposed to 3 doses of ethanol (6 g/kg bw) to induced fatty liver, while mice in control group received equal volume and equal energy of maltodextrin solution. NAC was administered to mice at 1 h before ethanol exposure (100 mg/kg bw, i.p.). The mice were sacrificed at 6 h after the last ethanol exposure. The liver and epididymal adipose tissues were collected. Histopathological examination and biochemical assay kit were used to evaluate the fat accumulation, while Western-blot was performed to detect the protein levels of some key factors involved in fat metabolism in liver and adipose tissues. Results: Compored with control group mice, the liver index and liver weight were significantly increased compared with model group, the liver index and TG level in NAC/ethanol group mice were all significantly decreased (P<0.05). Histological examination showed NAC effectively suppressed binge drinking-induced fat accumulation in mice liver. In addition, NAC had no significant effects on the protein levels of peroxisome proliferator-activated receptor-α (PPAR-α), Acy-CoA oxidase (ACOX), sterol regulatory element binding protein 1 c (SREBP-1c) and fatty acid synthase (FAS). Furthermore, the protein levels of hormone sensitive lipase (HSL) did not significantly differ among 3 groups, whereas NAC prevented binge drinking-induced increase of HSL phosphorylation at ser563 and ser660. Conclusion NAC could effectively attenuate binge drinking-induced fatty liver, which might be associated with the inhibition of lipid mobilization by suppressing the phosphorylation of HSL.

OBJECTIVE: To investigate the expression of caspase-3 during contused skeletal muscle wound healing in rats and to explore the applicability of caspase-3 in wound age estimation. METHODS: Male Sprague-Dawley rats undergone anesthesia with 2% pentobarbital sodium (45 mg/kg) were struck by a single strike at the skeletal muscle of the right leg, then were sacrificed by cervical dislocation at different post-traumatic intervals. The changes of caspase-3 activity were assessed by imunohistochemistry, Western blot and caspase-3 colorimetric assay. The skeletal muscle of non-contusion rat's were used as controls. RESULTS: No caspase-3 was detected in skeletal muscle in the control group. The expression level of caspase-3 in the peripheral area of contused rat skeletal muscle increased gradually and maximized on the fifth day after injury, and then decreased gradually. Caspase-3 was also detected in the regenerated skeletal muscle within 7-14 days after injury. CONCLUSION The expression of capase-3 in the peripheral area of contused rat skeletal muscle indicated that it may play a pivotal role in apoptosis of the wounded skeletal muscle and may be used as a marker for wound age estimation. The expression of capase-3 in regenerating skeletal muscle suggests that caspase-3 activity may be used for skeletal muscle differentiation.
Animals ; Caspase 3/metabolism* ; Contusions/enzymology* ; Forensic Pathology ; Male ; Muscle, Skeletal/injuries* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors ; Wound Healing