Objective To prepare the paliperidone long-acting suspension for injection and study the quality of it.Methods The optimum formula was measured by using orthogonal design method regarding the content of paliperidone, Na-CMC,mannitol and the ground time as variable factors. Orthogonal design table L9 ( 34 ) was used in the experiment. The paliperidone long-acting suspension for injection was prepared by grinding the dispersion,and its quality was evaluated. Results The optimum prescription was as follows: using paliperidone 40 g, Na-CMC 15 g, mannitol 60 g, purified water 1 000 mL, appropriate amount of NaH2 PO4 and NaOH, and grinding for 1 h. Paliperidone long-acting suspension for injection was characterized by an average pH of 7. 0,and ratio of average sedimentation volume of 90. 3%. The suspension released smoothly within 1 h in vitro and achieved sustained release. The suspension was stable in a freeze-dried powder. Conclusion The preparation for paliperidone long-acting suspension for injection is simple and stable,which may be a novel drug delivery system.

A 34-year-old male patient with suspected pheochromocytoma was referred to our department for paroxysmal hypertension. Pheochromocytoma was confirmed by very high plasma nor-metanephrine ( NMN ) and metanephrine ( MN ). Contrast CT of the adrenal disclosed a 3.0 cm x 3.0 cm mass in the left adrenal. 18 F-FDG-PET-CT showed high uptake only in the left adrenal. When the patient's blood pressure was well controlled with 4 mg/d doxazosin for 2 weeks, surgery was then performed under laparoscopy. A round solid left adrenal pheochromocytoma was resected. After surgery, plasma MN and NMN levels returned to normal, and the patient was free of clinical symptoms with normal blood pressure. This patient has been followed 3 years with normal blood pressure, MN, and NMN levels, without tumor recurrence shown hy adrenal CT.

Objective To make soluble expression of Plasmodium falciparum (FCC1/HN) glutamate dehydrogenase(GDH) in Escherichia coli, purification and immunocompetence identification of the recombinant non-fusion GDH. Methods The GDH gene was cloned into prokaryotic expression vector pET23(a) to form recombinant expression vector pET23(a)/GDH. pET23(a)/GDH was transformed into E.coli BL21(DE3). Induced by IPTG(isopropyl-beta D-thiogalactoside), GDH was highly expressed in the supernatant after sonication. The soluble recombinant GDH was purified by Source-Q and Source-S chromatography. Enzyme-linked immunosorbent assay and Western blotting were carried out to identify the immunocompetence of the purified product. Results SDS-PAGE analysis showed that the soluble GDH protein accounted for approximately 15% of the total bacterial protein. By two-step ion-exchange chromatography, the purity of GDH reached more than 90% and the GDH possessed high antigenicity. Conclusion The soluble expression of GDH results in an integral three-dimensional structure epitope with high biological activity.

Purpose To investigate the mutation status of KRAS and PIK3CA gene in colorectal cancer (CRC) primary lesions and corresponding liver metastasis and its clinical significance.Methods The gene mutations of KRAS and PIK3CA were detected in 58 cases of primary lesions of CRC and corresponding liver metastasis tissue by real-time PCR.Results The mutation rates of KRAS were 31.03％ (18/58) and 25.86％ (15/58) in primary lesions of CRC and corresponding liver metastasis tissue,respectively,in which G12D was most commonly detected.The mutation rates of PIK3CA were 8.62％ (5/58) and 10.34％ (6/58) respectively,in which the most common mutation site was E545K.Only one case carried simultaneously both mutations of KRAS (G12D) and PIK3CA (E545K).The mutation of KRAS and PIK3CA had a good consistency between primary lesions and liver metastasis.Univariate analysis showed that the mutation of KRAS was related to the primary lesion of tumor location,the quantity of metastasis and the types of tumor (P ＜ O.05),PIK3 CA mutation was associated with the synchronous/metachronous liver metastasis and the quantity of metastasis (P ＜ 0.05).Multivariate Cox regression analysis showed that synchronous/metachronous liver metastasis and the mutation of KRAS were influencing factors for prognosis of CRC.The overall survival of patients with CRC who had simultaneous liver metastases was longer than those with heterotopic liver metastases;the overall survival of KRAS wild-type mutant patients was longer than those of mutant patients (P ＜ 0.05).Conclusion The G12D site of KRAS gene has the highest mutation frequency in CRC,KRAS/PIK3CA mutation has a good consistency of the primary lesions of CRC and corresponding liver metastasis.Primary lesions can be as the source of molecular detection.To achieve individualized treatment,we need to reassess the genetic status of metastasis based on the choice of targeted therapy for precision medicine.

Objective To determine whether the CD133+ cancer stem cells lie in circumferential resection margin of rectal cancer,and to explore the relationship between CD133+ cancer stem cells and prognosis of rectal cancer.Methods The circumferential resection margin of rectal cancer was cut,one half of the tissue was stained HE for microscope observation.Both groups were labeled CD133 by immunohistochemistry.The tissues with positive circumferential resection margin were classified as experimental group,otherwise the control group.Another half was made into single cell suspension and used to detect CD133+ cancer stem cells by flow cytometry.The relapse of patients in the experimental group was following-up.Results Two cases were positive by immunohistochemistry in the experimental group (28 cases),showed that CD133+ cancer stem cells were in circumferential resection margin,but there was not marked difference compared with the control group (72 cases were all negative) (P =0.076).CD133+ cancer stem cells can be detected by flow cytometry,and the expression was higher in experimental group than that in the control group (5.19 ％ and 0.56 ％,Z =-7.739,P ＜ 0.01).In experimental group,the expression of CD133+ cancer stem cells in circumferential resection margin of relapse patients was significantly higher than that in patients without relapse,the difference was statistically significant [(6.57±1.72) ％ and (4.77±1.33) ％,t =3.038,P ＜ 0.05].Spearman rank correlation analysis showed that the proportion of CD133+ cancer stem cells in the experimental group was positively correlated with relapse (rs =0.473,P ＜ 0.05).Conclusion There are CD133 + cancer stem cells within circumferential resection margin of rectal cancer,and the expression of CD133+ cancer stem cells in experimental group is correlated with the postoperative recurrence.

Objective To investigate the significance of CXCL 12/CXCR4 expression in T lymphoblastic lymphoma/leukemia ( T-LBL/ALL ) and its prognostic significance . Methods Using immunohistochemical EnVision method , CXCL12, CXCR4 and Ki-67 expression were evaluated in 72 cases of T-LBL/ALL and 30 selected cases of lymph node reactive hyperplasia ( LH) as control.In addition, CXCL12 and CXCR4 mRNA expression levels were examined by real-time reverse transcription polymerase chain reaction ( real-time RT-PCR ) method.Results Immunohistochemical results showed that the expression rates of CXCL 12 and CXCR4 in T-LBL/ALL were 84.7%( 61/72 ) and 91.6%( 66/72 ) , respectively, and these were not different from the expression in the LH control group .The expression indexes of Ki-67 <80% and ≥80% were 25 cases (34.7%,25/72) and 47 cases (65.3%,47/72), respectively.Real-time quantitative PCR demonstrated that CXCL 12 and CXCR4 mRNA expression in T-LBL/ALL was 62.4%and 71.5%, respectively, and was statistically different (P<0.05) from that of the control group.Single factor analysis found that CXCL 12 mRNA expression in T-LBL/ALL was positively correlated with Ann Arbor staging and KPS score ( P<0.05 ); CXCL12 protein expression was positively correlated with splenomegaly ( P<0.05 ); CXCR4 mRNA expression was positively correlated with the IPI score, clinical symptoms, mediastinal widening and bone marrow involvement (P<0.05); CXCR4 protein expression was positively correlated with mediastinal widening ( P<0.05); CXCL12 mRNA expression was positively correlated with CXCL12 protein and CXCR4 protein expression (P<0.05), but not the CXCR4 mRNA and protein levels.There was no correlation between CXCL 12 and CXCR4 protein expression and CXCR4 mRNA expression.Multivariate COX regression analysis showed that high expression of CXCR 4 protein, hepatosplenomegaly and bone marrow involvement were risk factors for T-LBL/ALL outcome . Conclusions CXCL12/CXCR4 expression is associated with disease progress , mediastinal widening , bone marrow involvement and adverse outcome in T-LBL/ALL.CXCL12/CXCR4 axis plays an essential role in the occurrence and development of T-LBL/ALL.However , CXCL12 and CXCR4 protein expression are not entirely reflected by mRNA transcription levels , and there may be other molecules involved in CXCL 12/CXCR4 expression and regulation . With CXCR4 antagonists undergoing clinical trials , targeting the CXCL12/CXCR4 axis may be a promising treatment strategy for T-LBL/ALL.

OBJECTIVETo evaluate the clinical significance of No.12 lymph node dissection for advanced gastric cancer with D2 lymphadenectomy.

METHODSClinicopathologic data and No.12 lymph node dissection of 256 advanced gastric cancer patients undergoing radical operation in our department between January 2005 and December 2010 were retrospectively summarized and the influence factors of metastasis in No.12 lymph nodes were analyzed.

RESULTSOf 256 patients, 179 were male and 77 were female with the average age of 59.2 years. Tumor located in the upper of stomach in 24 cases, middle of stomach in 41 cases, lower of stomach in 174 cases, multi-focus or diffuse distribution of stomach in 17 cases. Tumor diameter was <3 cm in 39 cases, 3 to 5 cm in 100 cases, >5 cm in 117 cases. Serum carcinoembryonic antigen (CEA) level increased in 61 cases, serum carbohydrate antigens (CA)72-4 increased in 56 cases and CA19-9 increased in 61 cases. The number of No.12 lymph nodes resected from all the patients was 1 152, and the average number was 4.5±1.9. The metastasis rate of No.12 lymph nodes was 9.4%(24/256) after hematoxylin eosin staining (positive group). All the patients received effective follow-up to December 2015, and the average follow-up time was 101.2 months. The median survival time of positive No.12 group (24 cases) was 29.8 months and of negative No.12 group (232 cases) was 78.2 months, whose difference was statistically significant (χ=21.715, P=0.000). Univariate analysis found that No.12 lymph node metastasis was not associated with age, gender, tumor differentiation (all P>0.05), but was associated with tumor location, tumor diameter, invasive depth (all P<0.05), and was closely associated with Borrmann type, outside metastatic lymph nodes of No.12 and high levels of serum CEA, CA72-4 and CA19-9 (all P=0.000). Multivariate regression analysis found that tumor location (RR=2.452, 95%CI:1.537 to 3.267, P=0.000), Borrmann type (RR=1.864, 95%CI:1.121 to 3.099, P=0.016) and number of outside metastatic lymph nodes of No.12 (RR=2.979, 95%CI: 2.463 to 3.603, P=0.000) were the independent risk factors of the No.12 metastasis (P<0.05).

CONCLUSIONSMetastasis in No.12 lymph nodes indicates poorer prognosis. The No.12 lymph nodes of advanced gastric cancer patients with curative resection, especially those with the tumor located in the lower part, Borrmann type IIII(, outside metastatic lymph nodes of No.12, should be regularly cleaned.

Antigens, Tumor-Associated, Carbohydrate ; blood ; CA-19-9 Antigen ; blood ; Carcinoembryonic Antigen ; blood ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; diagnosis ; pathology ; physiopathology ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Grading ; statistics & numerical data ; Neoplasm Invasiveness ; Neoplasm Staging ; statistics & numerical data ; Prognosis ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; blood ; mortality ; pathology ; Survival Rate

OBJECTIVETo evaluate the feasibility, safety, radicality and short-term outcome of preserving left colic artery (LCA) during laparoscopic total mesorectal excision (TME) for the treatment of rectal cancer.

METHODSFrom January 2013 to December 2016,136 patients with mid-lower rectal cancer received laparoscopic TME in the Gastrointestinal Surgery Department of Taizhou People's Hospital of Jiangsu Province. Patients with rectal tumor within 10 cm to the anal verge were enrolled into the study. All the enrolled patients had complete data of pathology and follow-up. Those receiving neoadjuvant chemoradiotherapy, with severe base diseases, multifocal tumor, tumor invasion of surrounding tissues, fixation of tumor, recurrent tumor, complications such as acute ileus, bleeding, perforation were excluded. In this study, 72 patients did not undergo preservation of LCA (high ligation group) and 64 patients underwent preservation of LCA (low ligation group). Operative parameters, clinicopathological data and short-term outcome were collected and compared between two groups.

RESULTSThe baseline data including gender, age, body mass index, tumor stage, and distance of tumor from anal verge of two groups were comparable (P>0.05). The differences between two groups about the mean time of operation and the operative blood loss were not significant [(164.0±12.6) min vs. (167.3±9.4) min, (30.0±3.6) ml vs. (30.1±3.0) ml, all P>0.05]. There was no operative death in both groups. Differences in the lymph node dissection (13.7±2.6 vs. 13.3±2.1) and the specimen length of proximal resection margin [(16.4±1.9) cm vs. (16.7±2.1) cm] or distal resection margins [(3.9±0.6) cm vs. (4.1±0.9) cm] between high and low ligation groups were not significant (all P>0.05). Compared with high ligation group, the low ligation group had higher rate of sphincter preservation [92.2% (59/64) vs. 79.2% (57/72), χ=4.580, P=0.032], lower rate of anastomotic leakage [1.6% (1/64) vs. 9.7% (7/72), χ=4.075, P=0.044], anastomotic stenosis [3.1% (2/64) vs. 12.5%(9/72), χ=4.006, P=0.045], and voiding and sexual dysfunction [6.3%(4/64) vs. 18.1%(13/72), χ=4.317, P=0.038]. Mean time of follow-up was 19 months. In high ligation group, the local recurrent rate was 5.56%, distant metastasis rate was 13.89%, overall survival rate was 90.28%, disease-free survival rate was 80.56%, while in low ligation group, the local recurrence rate was 4.69%, distant metastasis rate was 12.50%, overall survival rate was 90.63%, disease-free survival rate was 82.81%, whose differences between two groups were not significant (all P>0.05).

CONCLUSIONPreservation of LCA during laparoscopic TME for the treatment of rectal cancer is safe and feasible, which can reduce the incidence of anastomotic leakage and stenosis, and voiding and sexual dysfunction.

OBJECTIVETo investigate clinicopathological features and prognostic factors of appendiceal neuroendocrine neoplasms(a-NEN).

METHODSClinical data of 20 patients diagnosed with a-NEN at Zhongshan Hospital of Fudan University between January 2000 and December 2016 were retrospectively analyzed. Pathological diagnosis was based on the WHO classification criteria of digestive system tumors (2010 edition). Based on the mitotic count and Ki-67 index, a-NENs were divided into grade 1 neuroendocrine tumor (NET G1), grade 2(G2) NET G2) and grade 3 (neuroendocrine carcinoma, NEC). Some special types of a-NEN (e.g. goblet cell carcinoid) and mixed adenoneuroendocrine neoplasms were classified as mixed adenoneuroendocrine carcinoma (MANEC). Follow-up was conducted by telephone or return visits. Univariate analysis was carried out using the Kaplan-Meier method, and the log-rank test was used to draw survival curves.

RESULTSOf 20 patients, 14 were male and 6 were female with median age of 54 years. Seventeen cases presented acute right lower quadrant abdominal pain, 1 chronic right lower quadrant abdominal pain, 1 persistent abdominal discomfort with outburst whole abdominal pain and 1 was found during body check without symptoms. Twenty cases comprised 8 G1 patients, 4 G2 patients, 3 G3 patients, and 5 MANEC patients. When diagnosed, there was 1 patient with liver metastasis, 1 patient with abdominal and pelvic metastases, and 2 patients with postoperative pathological findings of lymph node metastasis. Six patients underwent appendectomy, 12 underwent right hemicolectomy, 1 underwent right hemicolectomy plus small intestine resection, and 1 underwent partial hepatectomy plus right hemicolectomy. The follow-up time was 7-187 months(average, 36 months). The total 1- and 3-year survival rates were 94.7% and 60.2%, respectively. Univariate analysis showed that age >50 years (χ=7.036, P=0.008), pathology grade as MANEC (χ=5.297, P=0.021), and metastasis (χ=6.558, P=0.010) indicated lower 5-year survival rate.

CONCLUSIONSMost a-NEN patients have no typical symptoms, and the main complaint at consultation is acute right lower quadrant abdominal pain. Prognosis is poor for patients with age >50 years, MANEC pathology grade and metastasis.

Appendiceal Neoplasms ; complications ; diagnosis ; surgery ; Carcinoma, Neuroendocrine ; complications ; diagnosis ; therapy ; Female ; Gastrointestinal Neoplasms ; Humans ; Male ; Middle Aged ; Neuroendocrine Tumors ; complications ; diagnosis ; surgery ; Prognosis ; Retrospective Studies

OBJECTIVE: To evaluate the effect of rosmarinic acid (RA) on mitophagy and hypertrophy of cardiomyocytes exposed to high glucose (HG). METHODS: Rat cardiomyocytes (H9c2) exposed to HG (25 mmol/L) were treated with 50 μmol/L RA or with both RA treatment and Parkin siRNA transfection, with the cells cultured in normal glucose (5.5 mmol/L) and HG as the controls. The expressions of PINK1, Parkin and LC3II/LC3I in the cells were detected by Western blotting. The formation of mitochondrial autophagosomes was observed by transmission electron microscope. Flow cytometry was employed to detect the level of reactive oxygen species (ROS) and apoptotic rate of the cells. The activities of respiratory chain complex enzymes were measured by spectrophotometry. Fluorescence enzyme labeling and RESULTS: RA treatment significantly increased the expression levels of PINK1, Parkin and LC3-II/I ( CONCLUSIONS RA can protect rat cardiomyocytes against oxidative stress injury and cardiomyocyte hypertrophy induced by HG by activating Parkin-mediated mitophagy.
Animals ; Cinnamates ; Depsides ; Glucose ; Hypertrophy ; Mitophagy ; Myocytes, Cardiac ; Protein Kinases ; Rats ; Reactive Oxygen Species ; Ubiquitin-Protein Ligases/genetics*