OBJECTIVE: To determine the protective effects of nourishing spleen yin recipe (Zibu Piyin Recipe, ZBPYR), a compound traditional Chinese herbal medicine, on endoplasmic reticulum (ER) in neuronal cells responding to the stress by using sero-pharmacological method. METHODS: The mouse neuroblastoma cell line Neuro2a cells were treated with tunicamycin (Tm, an inhibitor of N-glycosylation). The ZBPYR-treated cell group was established by incubating cells with ZBPYR serum for one hour and treated with Tm. Reverse transcriptase PCR (RT-PCR) was utilized to detect the mRNA expressions from two genes after treatments, ER molecular chaperone glucose regulated protein 78 (GRP78) and transcriptional factor CCAAT/ enhancer-binding protein-homologous protein (CHOP). Lactate dehydrogenase (LDH) assay was also carried out to determine the LDH leakage from Neuro2a cells after treated with Tm and staurosporine (STS). RESULTS: The ZBPYR-treated cell group at all tested ZBPYR dosages showed significantly reduced expressions of both genes compared with Tm (5 microg/ml) treated control group (P<0.05). Therefore, ZBPYR serum inhibited the expressions of GRP78 and CHOP in mRNA level under ER stress induced by Tm. Different concentrations of ZBPYR serum pretreatment reduced the LDH leakage compared with the Tm and STS groups (P<0.05). Therefore, ZBPYR serum may inhibit the LDH leakage induced by Tm and STS. CONCLUSION: ZBPYR has neuroprotective effects. The mechanisms may be associated with inhibition of ER stress and mitochondrial dysfunction.

Background and Objective:The gross tumor volume (GTV) obviously reduces after induction chemotherapy (IC) for primary Iocoregionally advanced nasopharyngeal carcinoma (NPC). This study was to investigate the impact of changing gross tumor volume delineation on the dose distribution and clinical treatment outcome after IC. Methods: From January 2008 to April 2009, 24 patients with Stage Ⅲ-Ⅳb primary locoregionally advanced NPC were treated with TPF regimen IC followed by intensitymodulated radiotherapy (IMRT) with concurrent chemotherapy The primary GTVs were delineated into two parts: the post-IC primary GTV (GTVpost-IC-NP),and the region of pre-IC primary GTV minus GTVpost-IC-NP (GTVprepost-IC-NP). The dose distributions of two plans with GTVpost-IC-NP or pre-IC primary GTV were assessed by analyzing ten cases. The clinical treatment outcome and toxicity of all patients were observed. Results:The post-IC GTV was significantly smaller than the pre-IC GTV(primary GTV 25.5 cm~3 vs.51.1 cm~3,P=0.001;lymph nodes GTV 9.1 cm~3 vs. 31.4 cm~3, P=0.035;primary+lymph nodes GTV 33.2 cm~3 vs. 82.6 cm~3, P=0.004), the overall GTV with an average shrinkage of 61%. The high dose region was also smaller after IC(volumes covered by 64.4 Gy were 422.9 cm~3 vs.457.9cm~3, P=0.003; 274.2 cm~3 vs.334.5 cm~3 by 68 Gy, P=0.041). The complete response rate was 38% after IC,and 100% three month after radiotherapy.The toxicity of following IMRT with concurrent chemotherapy was similar to that of IMRT with concurrent chemotherapy alone. With median follow-up of 9 months, the locoregionally control rate was 100% and only one patient presented metastasis 15 months after treatment. Gonclusions: TPF regimen IC could significantly reduce tumor volume. The following IMRT with GTVpost-IC-NP plan reduced the high dose region,which didn't add toxicity while had excellent short-term treatment outcome.

OBJECTIVETo improve the accuracy of preoperative evaluation of cervical lymph node metastasis in thoracic esophageal squamous carcinoma.

METHODSForty-two patients with thoracic esophageal squamous carcinoma underwent neck ultrasonography. Enlarged lymph nodes with their long axis greater than 10 mm and a short-to-long axis ratio greater than 0.5 were considered as metastatic.

RESULTSPreoperative neck ultrasonography revealed the enlarged lymph nodes in 16 patients, but only in 5 (31%) cases the nodes were palpable. Among them 9 were classified as metastatic (cM(1-LN)), including 4 patients with palpable nodes. In 5 cM(1-LN) patients surgical intervention was canceled and the remaining 37 patients underwent trans-thoracic esophagectomy. Cervical node metastasis (pM(1-LN)) was confirmed pathologically in 6 surgical patients, 4 with tumors invading the adventitia (pT3) and the other 2 into the surrounding structure (pT(4)) (pT(1), pT(2) vs. pT(3), pT(4), P = 0.020). All 6 pM(1-LN) patients had concomitant mediastinal node metastasis and 4 of them had upper abdominal node metastasis. Statistically significant relationship was detected between cervical and abdominal nodal status (r = 0.536, P = 0.007). In comparison with the results of pathological examination and treatment response, the accuracy and sensitivity were 81% and 95% (P = 0.043), 36% and 82% (P = 0.081), respectively, for palpation and ultrasonography. Five out of 39 (13%) patients had their therapy changed due to ultrasonographic findings.

CONCLUSIONSNeck ultrasonography for cervical lymphadenopathy is of high sensitivity and accuracy, which plays an important role in the preoperative evaluation and therapeutic decision-making.

Adult ; Aged ; Carcinoma, Squamous Cell ; diagnostic imaging ; secondary ; surgery ; Esophageal Neoplasms ; pathology ; surgery ; Female ; Head and Neck Neoplasms ; diagnostic imaging ; secondary ; surgery ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; diagnostic imaging ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Neck ; diagnostic imaging ; Sensitivity and Specificity ; Ultrasonography

OBJECTIVETo investigate the pathological diagnosis, surgical treatment and prognostic factors of gastrointestinal stromal tumors (GISTs).

METHODSThe clinicopathological data of operated 96 patients with GISTs were analyzed retrospectively. Expression of CD117, CD34, SMA and S-100 was determined by immunohistochemical methods.

RESULTSExpression of CD117, CD34, SMA and S-100 was 79.2% (76/96), 58.3% (56/96), 35.4% (34/96) and 9.4% (9/96). Benign tumor 23 and malignant 73. Of the malignant, the omentum was resected in 39 and the rest remained, of which the recurrent and metastatic rates were 5.1% and 26.5% (P < 0.05). The safety margin between the normal intestine and tumor was > 5 cm in 46 patients; while in the other 27 patients, it was < 5 cm. The recurrent and metastatic rates were 6.5% and 29.6% (P < 0.05), respectively. The 5-year survival rates of benign and malignant GISTs were 91.5% and 57.3% (P < 0.05).

CONCLUSIONThe application of immunohistochemical markers CD117 and CD34 are supplementary to pathological diagnosis. The adapting of rational primary treatment, including complete tumor resection and prophylactic omentectomy, is able to reduce the recurrence of GISTs.

Adolescent ; Adult ; Aged ; Antigens, CD34 ; metabolism ; Child ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; diagnosis ; metabolism ; pathology ; surgery ; Humans ; Liver Neoplasms ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Omentum ; surgery ; Peritoneal Neoplasms ; secondary ; Proto-Oncogene Proteins c-kit ; metabolism ; Retrospective Studies ; Survival Rate

Objective To discuss the anatomic features, clinical presentations, diagnosis,differentiations and treatments of congenital fourth branchial anomaly(CFBA). Methods The clinical data of 8 patients with CFBA were retrospectively analyzed. Results Of the 8 patients aging from 27 to 300 months(median age: 114 months), 4 male and 4 female; 3 untreated previously and 5 recurrent. All lesions, including 1 cyst, 3 sinus (with internal opening) and 4 fistula, located in the left necks. Three patients presented acute suppurative thyroiditis, 4 deep neck abscesses, and 1 neck lump. Preoperative examinations included barium esophagogram, direct laryngoscopy, ultrasonography, CT, MRI, and so on.The principles of managements were adequate drainage, infection control during acute period and radical surgery during quiescent period. Classic surgical approach consisted of complete excision of branchial lesions, dissection of recurrent laryngeal nerve and partial thyroidectomy. Selective neck dissection was applied in recurrent cases to extirpate branchial lesions, scarrings and inflammatory granuloma.Postoperatively, 1 case was with local incision infection which healed by wound care; 1 case was with temporary vocal cord paralysis which completely recovered 1 month after operation. No recurrence was found in all of 8 cases with follow-up of 13 to 42 months (median: 21 months). Conclusions CFBA relates closely anatomically with recurrent laryngeal nerve and thyroid grand. The barium esophagogram and direct laryngoscopy are the most useful diagnostic tools. CT and MRI are all beneficial to the diagnosis of CFBA.The treatment key to CFBA is the complete excision of lesion during a quiescent period after inflammatory control, together with the dissection of recurrent laryngeal nerve ,partial thyroidectomy and partial resection of lamina of thyroid cartilage (if necessary), which all can decrease the risk of complications and recurrence.For recurrent cases, selective neck dissection is a safe and effective surgical procedure.

AIMTo study the microbial transformation of sinenxan A.

METHODSChoose two strains of Fungi (Mucor spinosus AS 3.3450 and Cunninghamella echinulata AS 3.3400) and a strain of bacterium (Proteus vulgaris AS 1.1208) to transform the substrate.

RESULTSThree products were obtained and identified as 10-deacetylsinenxan A1, 6 alpha-hydroxy-10-deacetylsinenxan A2 and 9 alpha-hydroxy-10-deacetylsinenxan A3 respectively.

CONCLUSIONSinenxan A is facile to be transformed by microorganisms, the 10-acetyl group of which is an active group.

Acetates ; isolation & purification ; metabolism ; Biotransformation ; Culture Techniques ; Cunninghamella ; metabolism ; Diterpenes ; isolation & purification ; metabolism ; Mucor ; metabolism ; Plants, Medicinal ; chemistry ; Proteus vulgaris ; metabolism ; Taxus ; chemistry

OBJECTIVETo prepare nanoparticles containing E1A gene and observe the efficiency and feasibility of transfecting E1A gene into human undifferentiated thyroid cancer cell line HTC/3. To examine the sensitivity of transgene cells to X-ray and X-ray-induced apoptosis in those cells.

METHODSNanoparticle-DNA complex was prepared with PLGA coating adenoviral early expression gene E1A, and the package efficiency, release progress in vitro, and size of the complex were determined. The nanoparticle-DNA was transfected into the HTC/3 cells. Lipofectamine was used to transfect E1A gene as a control. RT-PCR was used to examine E1A gene mRNA expression in the transfected cells. The survival ratio of HTC/3-E1A and control cells, and the growth inhibition ratio induced by different doses of X-ray in HTC/3-E1A cells were examined by MTT assay. The apoptosis in HTC/3-E1A cells induced by 2 Gy X-ray iradiation was examined by flow cytometry and DNA electrophoresis.

RESULTSThe package efficiency, release progress in vitro, and size of the nanoparticle-DNA complex were 0.78%, 18 days, and 150-280 nm, respectively when transfected the plasmid at the same level, the nanoparticle group got more positive transgene cell clones than that in lipofectamine group, with a statistically significant difference (P < 0.05). RT-PCR showed that transgenic cells from both nanoparticle-DNA and lipofectamine groups had E1A gene mRNA expression. The HTC/3-E1A cells grew slowly, and their doubling time was prolongated (1.44 times in comparison with that in parental cells). According to IC50, the sensitivity of HTC/3-E1A cells to X-ray was improved 2.9 and 2.8 times, respectively, in comparison with that in HTC/3-Vect and HTC/3 cells. The ratio of subG0/G1 phase of HTC/3-E1A cells was significantly higher than that in HTC/3-Vect and HTC/3 cells (P < 0.01). The ratio of S phase of HTC/3-E1A cells was significantly lower than that in HTC/3-Vect and HTC/3 cells (P < 0.01). A typical DNA ladder pattern of apoptosis in HTC/3-E1A cells was observed by electrophoresis, but not found in HTC/3-Vect and HTC/3 cells.

CONCLUSIONA nanoparticle-DNA complex has been successfully prepared, and it may carry a foreign gene into cells. The sensitivity of HTC/3-E1A cells to X-ray is significantly improved. Moreover, apoptosis is induced by x-ray in the E1A gene-transfected cells.

Adenovirus E1A Proteins ; biosynthesis ; genetics ; physiology ; Apoptosis ; radiation effects ; Cell Cycle ; radiation effects ; Cell Line, Tumor ; Cell Proliferation ; DNA ; genetics ; Humans ; Lactic Acid ; chemistry ; Nanoparticles ; Particle Size ; Plasmids ; Polyglycolic Acid ; chemistry ; RNA, Messenger ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology ; Transfection ; X-Rays

OBJECTIVETo observe the influence of recombinant adenovirus-mediated PDE5-shRNAs on the free calcium level in rat penile smooth muscle cells and to explore the feasibility of gene therapy for erectile dysfunction (ED).

METHODSSmooth muscle cells of the rat corpus cavernosum were transfected with constructed rAd-rPDE5-shRNAs and then dyed with the calcium fluorescent probe Fluo-3/AM at 24, 48 and 72 hours. The dynamic changes of the calcium fluorescence intensity were observed under the laser scanning confocal microscope (LSCM). The relative level of intracellular calcium was determined by fluorescence indexes.

RESULTSThe fluorescence indexes of calcium at 24, 48 and 72 hours were 829.3 +/- 7.8, 801.5 +/- 9.5 and 856.3 +/- 8.7 in the rAd-rPDES-shRNAs group, significantly lower than in the rAd-mock (1106.3 +/- 10.8, 1121.3 +/- 10.2 and 1058.5 +/- 12.1) and blank control group (1076.6 +/- 9.7, 1133.4 +/- 11.2 and 1104.3 +/- 10.5) (P < 0.05).

CONCLUSIONAdenovirus mediated shRNAs of the target PDE5 gene can significantly decrease the intracellular calcium level in the smooth muscle cells of the rat corpus cavernosum.

Adenoviridae ; genetics ; Animals ; Calcium ; metabolism ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; metabolism ; Erectile Dysfunction ; therapy ; Gene Silencing ; Genetic Therapy ; Male ; Myocytes, Smooth Muscle ; metabolism ; Penis ; cytology ; RNA, Small Interfering ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of phosphodiesterase type 5 (PDE5) small interfering RNA (siRNA) on cyclic guanosine monophosphatethe (cGMP) in the smooth muscle cells of human corpus cavernosum, and to provide laboratory evidence for the application of the RNA interference (RNAi) technique for the treatment of erectile dysfunction.

METHODSThe recombinant adenovirus rAd5-shRNA-PDE5A3 expressing three pairs of specific shRNA was constructed successfully. The smooth muscle cells of human corpus cavernosum were divided into an experimental, a negative control and a blank control group, and transfected respectively with rAd5-shRNA-PDE5A3, adenovirus rAd5-mock and phosphate buffered saline. The concentration of cGMP was measured by radioimmunoassay at 24, 48 and 72 hours after transfection, and the effect of rAd5-shRNA-PDE5A3 was detected on the cGMP in the smooth muscle cells of the corpus cavernosum.

RESULTSThe cGMP level in the smooth muscle cells of the corpus cavernosum was significantly higher in the rAd5-shRNA-PDE5A3 group than in the rAd5-mock control and blank control groups (P < 0.05), most significantly at 72 hours after transfection.

CONCLUSIONThe rAd5-shRNA-PDE5A3 can obviously increase the cGMP level in the smooth muscle cells of human corpus cavernosum, and enhance the inhibition of the PDE5 gene.

Adenoviridae ; Cells, Cultured ; Cyclic GMP ; metabolism ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; genetics ; Erectile Dysfunction ; genetics ; Genetic Vectors ; Humans ; Male ; Myocytes, Smooth Muscle ; metabolism ; Penis ; cytology ; RNA, Small Interfering

OBJECTIVETo observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells.

METHODSSDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co-cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control.

RESULTSThe level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively.

CONCLUSIONDown-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells.

Bone Marrow Cells ; metabolism ; Cell Adhesion ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; metabolism ; Coculture Techniques ; Drug Resistance, Neoplasm ; Gene Expression ; Humans ; Jurkat Cells ; RNA Interference ; Stromal Cells ; metabolism