Objective To investigate the effects of apoptosis and related gene p21WAF1 and Bcl-2 in the tumorigenesis of primary gallbladder neoplasms. Methods p21WAF1 and Bcl-2 were detected by immunohistochemistry and apoptotic cells were stained in situ by terminal deoxynucleotidyl transferase mediated-dUTP nick end labeling(TUNAL) in 46 cases of primary gallbladder neoplasms with different histological grades. Results Apoptotic index and p21WAF1 were decreased,but the expression of Bcl-2 was increased with the increasing of the histological grades in primary gallbladder neoplasms (P

To observe the curative effect of treating small -incision cataract extraction by intraocular lens implantation combined with trabeculectomy for primary angle-closure glaucoma with cataract.
●METHODS: Totally 44 cases (52 eyes) of primary angle-closure glaucoma combined with cataract were selected to undergo the combined surgery, in order to observe the patients' pre - and postoperative eyesights, intraocular pressures and the postoperative complications.
●RESULTS: The postoperative eyesight was improved significantly as compared with the preoperative eyesight. The intraocular pressure was declined dramatically. The result was of statistical significance (P<0. 05). All the 52 cases' surgeries were performed by the same surgeon. The surgeries were processed smoothly, with 6 postoperative eyes of anterior chamber inflammation cell response, 3 eyes of anterior chamber fibrinoid exudate, 2 eyes of shallow anterior chamber through mydriasis and treatment with glucocorticoids and non - steroidal eyedrops before absorption, and no complications like malignant glaucoma, cyclodialysis, etc. were reported through mydriasis and pressure bandaging before recovery.
● CONCLUSlON: Treating the primary angle - closure glaucoma combined with cataract through the combined surgery has high reliability and desirable curative effect. The surgical method is simple to learn and applicable for promotion on the basic level.

Objective To establish a simple diabetic peripheral neuropathy (DPN) rat model with the high fat-fed in GK rats. Methods A total of 30 GK rats (7-8 weeks) were fed with high-fat diet to establish the DPN model. Thirty normal Wistar rats were fed with ordinary diet (control group). The blood-sugar value, body mass, water-intake and food-intake were monitored every week in two groups. The serum level of glycosylated hemoglobin, the right sciatic nerve conduction velocity were detected at 8, 12 and 16 weeks respectively. The left sciatic nerve was used for HE and TUNEL staining. Results The manifestations of polydipsia, polyphagia and growth retardation were gradually appeared in GK rats. After 12 and 16 weeks, the blood-sugar and glycosylated hemoglobin were significantly increased in GK rats compared with those of normal Wistar rats (P < 0.01). The sensory nerve conduction velocity decreased obviously (P < 0.01). And motor nerve conduction velocity showed a certain decline trend (12 week P < 0.05,16 week P > 0.05). The sciatic nerve pathological features and Schwann cell apoptosis suggested that the model of DPN was successfully established (apoptosis index, P <0.01). Conclusion GK rats fed by high-fat diet are the satisfactory models of the DPN in experimental research. And 12-week is a suitable and economical time for molding.

OBJECTIVETo establish genetic method in detecting Cryptosporidium parvum and Giardia lamblia which often coinfected with AIDS patients.

METHODSCryptosporidium oocysts and Giardia cysts were isolated and purified from fecal samples of the individuals infected with C. parvum and G. lamblia, respectively. Genomic DNAs were extracted. Two pairs of specific primers were designed or synthesized according to the 18S rRNA gene from C. parvum or the triose phosphate isomerase (tim ) gene from G. lamblia. Polymerase chain reaction(PCR) technique was used to amplify the DNA samples from the oocysts and the cysts, and those from the 6 control samples, including Schitosoma japonicum, Toxoplasma gondii , Entamoeba histolytica, Trichinella spiralis, Trichomonas vaginalis and human blood cells. DNA samples from 30 fecal samples of AIDS patients were detected with the same method.

RESULTSOne fragment of 500 bp was amplified with the primer of C. parvum, and the other one of 683 bp was amplified with the primer of G. lamblia. Twenty pg and 0.4 pg DNA of C. parvum and G. lamblia could be detected separately. The specificity of these two pairs of PCR primers was confirmed by the failure in the amplification of the control DNA samples. Out of 30 cases of AIDS patients, 7 showed C. parvum positive, while non Giardia was detected.

CONCLUSIONGenetic detection method for C. parvum and G. lamblia detection was established which was more sensitive and specific.

Acquired Immunodeficiency Syndrome ; microbiology ; Cryptosporidiosis ; diagnosis ; Cryptosporidium parvum ; genetics ; DNA, Bacterial ; Giardia lamblia ; genetics ; Giardiasis ; diagnosis ; Humans ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo study the possible anti-apoptotic mechanism of ginsenoside Rg1 on the apoptosis of hippocampal neuron after cerebral ischemia/reperfusion (I/R) injury rats.

METHODSTotally 120 healthy male adult SD rats were randomly divided into the cerebral I/R model group (the model group), the low dose ginsenoside Rg1 group (10 mg/kg), the middle dose ginsenoside Rg1 group (20 mg/kg), the high dose ginsenoside Rg1 group (40 mg/kg), and the sham-operation group, 18 in each group. Rats received medication by peritoneal injection. Equal volume of normal saline was peritoneally injected to rats in the sham-operation group and the model group, once daily, for 7 successive days. The cerebral I/R injury model was prepared by 2-h middle cerebral artery occlusion (MCAO) followed by 24-h reperfusion. Rats in the sham-operation group received the same surgical procedure without the carotid arteries occluded. The neurofunction was assessed using Longa EZ method. The injury of hippocampal pyramidal cells was observed by Nissel staining and TUNEL assay. The nerve cell apoptosis rate was calculated. The protein expression levels of extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2), c-Jun N-terminal kinases (JNK), and phosphorylated c-Jun N-terminal kinase (p-JNK) were detected using Western blot.

RESULTSCompared with the sham-operation group, the score of neurofunction, the apoptosis rate, the expression levels of p-JNK and p-ERK1/2 increased, the survived number of pyramidal cells decreased in the model group (P < 0.05, P < 0.01). Compared with the model group, the score of neurofunction and the apoptosis rate decreased in each ginsenoside Rg1 group (P < 0.05, P < 0.01). The survived number of pyramidal cells increased in the high and middle dose ginsenoside Rg1 groups, the expression of p-JNK in the hippocampal CA1 region decreased, and the expression level of p-ERK1/2 increased (P < 0.05, P < 0.01). Compared with the low dose ginsenoside Rg1 group, the score of neurofunction, the apoptosis rate, the p-JNK protein expression decreased, the survived number of pyramidal cells increased, the expression of p-ERK1/2 increased in the high and middle dose ginsenoside Rg1 groups (P < 0.05, P < 0.01). Three to four layers of pyramidal cells were arranged tightly and compactly in the hippocampal CA1 region of the sham - operation group. The nucleus was big and round under high power lens, with 1 -2 kernel. After cerebral I/R injury, the hippocampal nerve cells were severely injured. Normal structure was lost in the CA1 region, with disarranged cell line and reduced cell amount. Partial neurons were shrunken, and the kernel was condensed and darkenedly stained. They were in triangular, long strip, fusiform, or irregular shape. The staining of nucleus was clustered and the kernel was not clear. Ginsenoside Rg1 (20 and 40 mg/kg) could improve the morphology of ischemic nerve cells, reduce their loss. Of them, stronger effects were shown in the high dose ginsenoside Rg1 group than in the middle dose ginsenoside Rg1 group. The JNK protein band was divided into two subzones, JNK1 (46 kD) and JNK2 (54 kD). ERK protein band was also divided into two subzones, ERK1 (44 kD) and ERK2 (42 kD).

CONCLUSIONThe protective effect of ginsenoside Rg1 on cerebral I/R injury was correlated with inhibiting the apoptosis of hippocampal neurons, regulating the expression levels of p-ERK1/2 and p-JNK.

Animals ; Brain Ischemia ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Ginsenosides ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

OBJECTIVETo examine the effect of trastuzumab on cell proliferation, colony formation and changes of HER-2 proteins in human breast cancer cell line SKBR3 and human ovarian cancer cell line SKOV3 cells which overexpress p185 HER-2 but shed high or low HER-2 extracellular domain (ECD) levels.

METHODSSKBR3 cells and SKOV3 cells were treated with or without trastuzumab. Cell number and the rate of colony formation were calculated. Western blot analysis was used to detect p185 HER-2, HER-2 ECD and phospho-HER-2. Two-site ELISA assay was used for the detection of HER-2 ECD.

RESULTSTrastuzumab inhibited cell proliferation, colony formation, and decreased or eliminated the levels of two uncharacterized phospho-proteins (molar weight about 90 000 and 40 000) in SKBR3 cells shedding high level of HER-2 ECD expression. These responses were not observed in SKOV3 cells shedding low level of HER-2 ECD expression. But total p185, phospho-p185 and phospho-p95 proteins did not appear to change in SKBR3 and SKOV3 cells after treatment with trastuzumab. Trastuzumab reacts not only with proteolytic cleavage HER-2 ECD containing HER-2 ECD I , II , III and IV subdomains of p185 HER-2 extracellular domain, but also with the secreted autoinhibitor p68/ECD III a specifying 340 residues, identical to subdomains I and II from the extracellular domain of p185 HER-2, followed by a unique C-terminal sequence of 79 aa encoded by intron 8, which suggested that there may be a trastuzumab binding site on p68/ECD III a protein. Comparing with HER-2 ECD levels of the same number of SKBR3 cells, there was no significant decrease of HER-2 ECD shedding level after treatment with or without trastuzumab for 4 days in serum-free medium.

CONCLUSIONAntitumor effects of trastuzumab may be related to the two uncharacterized phospho-p90 and/or phospho-p40 proteins. There is probably a trastuzumab epitope on p68/ECD III a. The decrease of HER-2 ECD levels may be positively correlated with the number of SKBR3 cells.

Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Ovarian Neoplasms ; metabolism ; pathology ; Phosphorylation ; Receptor, ErbB-2 ; metabolism ; Trastuzumab

The study established a UPLC-MS/MS method that is used for simultaneous determination nine major bioactive compounds of Dachengqi Tang in rat plasma. Using Aglient C18 column (2.1 mm x 50 mm,1.7 microm) was chromatographed, using methanol-5 mmol x L(-1) ammonium formate mobile phase gradient, elution 0.3 mL x min(-1). In the plasma pre-treatment process, not only the method of methanol and acetonitrile protein precipitation was investigated, and different factors extraction solvent, the type of the scroll time, the number and the type of extraction solvent, the extraction volume of the extraction solution of liquid-liquid extraction is investigated. Finally, with ibuprofen as an internal standard, using ethyl acetate liquid-liquid extraction method pretreatment blood, N2 dry reconstituted supernatant after centrifugation UPLC-MS/MS analysis, in electrospray ionization (ESI) negative mode, using multiple reaction monitoring mode for testing. The linear range of emodin, rhein, aloe-emodin, chrysophanol, magnolol, honokiol, hesperidin and hesperitin is 0.33-660, 0.40-792, 0.41-827, 0.34-680, 0.45-907, 0.46-927, 0.43-867, 0.34-683, 0.39-787 microg x L(-1) respectively, good linear relationship; and extraction recovery were greater than 69.39%, days after the day of the RSD is less than 15%. This method can be used to study the rat gastric large bearing gas after Dachengqi Tang, the simultaneous determination of nine components in plasma for its pharmacokinetics and efficacy material base to provide a theoretical basis.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

OBJECTIVETo analyze the degrees on the epidemic foci of Angiostronglus cantonensis and to explore the measurement methods.

METHODSSnails (Pila gigas) were collected from the spots of Wenzhou, Cangnan, Yongjia, Yueqing in Zhejiang province and Minhou, Changle, Ningde in Fujian province. The snails were examined microscopically in order to calculate their infection rates and the average worm number in the positive snails, then taking the product of multiplication of both values as infestation index.

RESULTSThe infection rates of the epidemic foci were 10.59% (9/85), 60.74% (181/298), 34.96% (79/226), 32.90% (76/231), 57.50% (184/320), 40.00% (82/205), 17.65% (12/68) and the rates of infectivity were 6.57, 183.54, 121.73, 93.45, 276.36, 76.08, 12.65, respectively.

CONCLUSIONThe epidemic foci were divided into five ranks (super, high, mid-range, low and non-epidemic foci) according to the value of infestation index which ranked from > 75, 30-75, 5-29, < 5 to 0.

Angiostrongylus ; pathogenicity ; Animals ; China ; Disease Outbreaks ; Snails ; parasitology ; Strongylida Infections ; epidemiology

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA

Objective To analyze the prognostic factors of patients with adenocarcinoma of gastroesophageal junction (AGEJ) after radical resection. Methods In this retrospective study, 269 patients with AGEJ who underwent curative resection from March 2004 to June 2007 were enrolled.The survival curve was drawn by Kaplan-Meier method and the survival analysis was done by Log-rank test. Univariate and multivariate analyses of the prognostic factors of AGEJ were made by Cox model. Results The overall 5-year survival rate of 269 patients was 25.0 % with the median survival time of 22 months.The Siewert type and the number of positive lymph nodes could influence the survival rate (both P < 0.05). Among the 269 patients, 216 (80.3 %) were Siewert Ⅱ type, their median survival time was 30 and 12 months in the thoracic surgery group and the laparotomy group when the tumor diameter was 3-7 cm, the difference was statistically significant (χ2= 5.036, P= 0.025). Univariate analysis showed that tumor diameter, pT, pN, operation time, age and sex were significantly associated with survival rate (all P < 0.05). Cox multivariate analysis showed that patients with a more advanced tumor size suffered from a poorer prognosis (P< 0.05). The risk of postoperative death in patients with lymph node metastasis was 1.854 times that of patients without lymph node metastasis. Conclusions Patients with AGEJ are predominantly Siewert Ⅱ type, the transthoracic approach or thoracoabdominal approach are reasonable selections for these patients. Tumor diameter and pN are independent prognostic factors of AGEJ patients.