C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

OBJECTIVETo investigate magnetic resonance imaging(MRI) manifestations of perianal abscess and to evaluate MRI diagnosis for perianal abscess.

METHODSFifty cases of perianal abscess between July 2007 and March 2009 were included in this study. MRI was performed using T1 weighted sequence in axial plane, T2 weighted sequence with fat saturation in axial, coronal and/or sagittal plane, and T1 weighted enhanced sequence with fat saturation in axial, coronal and/or sagittal plane. The location, size, shape, signal characteristics, and enhancement patterns of perianal abscess were analyzed.

RESULTSFifty cases with 51 perianal abscesses were identified. The abscess appeared slight hypo- or isointense signal on T1 weighted imaging, obviously hyperintense signal at cavity and isointense signal at wall on T2 weighted with fat saturation imaging, and marked enhancement at wall. The largest diameters ranged between 0.5 cm and 9.0 cm (mean 3.4±1.7 cm ) in abscess, between 0.2 cm and 8.0 cm(mean 2.7±1.7 cm) in cavity. The abscess was round, oblong and crescent in 23, 18 and 7 cases respectively, and was unilocular in 41 cases and multilocular in 10 cases. The abscess located between levator ani and intersphincteric groove in 23 cases, under intersphincteric groove in 3 cases, across levator ani and intersphincteric groove in one case, on levator ani in 7 cases, on intersphincteric groove in 16 cases and above levator ani in one case.

CONCLUSIONMRI is a noninvasive, convenient and highly accurate procedure for diagnosing perianal abscess, which can demonstrate the anatomical relationship between abscess and anal canal.

Abscess ; diagnosis ; pathology ; Adolescent ; Adult ; Aged ; Anus Diseases ; diagnosis ; pathology ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

An anti-alpha-synuclein (alpha-SYN) monoclonal antibody produced in our laboratory was used to investigate the effect of repeated acute hypoxic treatments on the expression of alpha-SYN in the mouse cerebral cortex. Western blot analysis showed that the expression levels of alpha-SYN in the cortex changed accordingly upon hypoxic exposure times, as that the alpha-synuclein level significantly increased after the first hypoxic exposure and then dropped down to the background level after the fourth hypoxic exposure. Immunohistochemical staining revealed that the alpha-SYN-immunopositive substance was localized not only in the nerve endings, but also within the nuclei of some neurons. The cell density of the neurons with alpha-SYN immunopositive nuclei was increased significantly after the first hypoxic exposure but returned back to control levels after the fourth hypoxic exposure. Our results indicate that both of the alpha-SYN expression level in the brain and the number of the neurons with alpha-SYN positive nuclei are affected by the repeated acute hypoxic treatments and that this modification is hypoxic time-dependent. The mechanism and the physiological significance underlying these changes need to be further investigated.
Animals ; Brain ; blood supply ; Brain Ischemia ; metabolism ; Cerebral Cortex ; metabolism ; Ischemic Preconditioning ; Mice ; Mice, Inbred BALB C ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Neurons ; metabolism ; Phosphoproteins ; biosynthesis ; genetics ; Random Allocation ; Synucleins ; alpha-Synuclein

Objective To investigate the clinical features of recurrent epileptic seizures and perioperative management principles and methods of neurosurgical patients with recurrent epileptic seizures.Methods The clinical data,reasons for aggravated seizures,seizure characteristics,and treatment methods and results of 9 patients with recurrent epileptic seizures were analyzed retrospectively.Results Of all the 9 patients,3 were combined with glioma,1 with arachnoid cyst,1 with cavernous hemangioma and 1 with encephalomalacia; epilepsy history was noted in 7 patients; frontal lobe epilepsy was noted in 7,and temporal lobe epilepsy in 2.The reasons for seizure aggravation included drug-decrement(n=3),recent-diagnosed brain tumor(n=2)and surgical operation during intracranial electrode implantation(n=1); and unknown reason was noted in the other 3.Epileptic seizure types included partial seizure and secondary generalized seizure; the frequency of seizure ranged from 3 minutes of interval to several hours of interval.Patients given multiple antiepileptic drugs,including oral and parenteral administration,received good seizure-control; and levetiracetam showed good curative effect during the treatment.Conclusion Recurrent seizures show drug refractory,and is hard to control.The AEDs having good curative effect in partial seizure should be combined used in these patients with higher dosage than conventional initial dose; intravenous and intramuscular administration can be used; the seizures should be controlled as soon as posible.Levetiracetam has good curative effect on patients with recurrent seizures for its fast oral-absorption,rapid onset of action and good antiepileptic effect.

OBJECTIVETo study the impacts of HIV/HCV co-infection to NK cells by investigating the changes of frequencies and functions of NK cells in HIV/HCV co-infected patients.

METHODSFrequencies and counts of NK cells were measured in patients with HIV mono-infection, HCV monoinfection, HIV/HCV co-infection and health control (HC) group by flow cytometer (FCM). After stimulated with PMA and K562 cells, PBMCs were examined the proportion of IFN-gamma+ NK cells by FCM. Proportion of killed K562 cells were detected to analyze the killing functions of NK cells.

RESULTSThe frequencies of NK cells, the percentages of IFN-gamma+ NK cells as well as the functions of NK cells killing the K562 cells in HIV/HCV co-infection, HIV mono-infection and HCV mono-infection groups were all lower than those of HC group significantly, the absolute counts and killing functions of NK cells in co-infection group were significantly lower than those of HIV or HCV mono-infection group.

CONCLUSIONSThe counts and functions of NK cells were affected more in HIV/HCV co-infections than those in HIV or HCV mono-infection.

Case-Control Studies ; Cell Count ; Coinfection ; immunology ; Flow Cytometry ; HIV Infections ; immunology ; Hepatitis C ; immunology ; Humans ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology

OBJECTIVETo explore a more optimal therapy for intractable insomnia.

METHODSSeven hundred cases of intractable insomnia that were in accordance with the criteria were randomly divided into an observation group (368 cases) and a control group (332 cases). The acupuncture prescription of regulating yin-yang and five viscera was applied in the observation group, where Dazhui (GV 14), Shenmai (BL 62), Guanyuan (CV 4), Zhaohai (KI 6), Geshu (BL 17), etc. were selected. The acupuncture prescription of tranquilizing mind was applied in the control group, where Baihui (GV 20), Sishencong (EX-HN 1), Anmian (Extra), Shenmen (HT 7), Sanyin jiao (SP 6) were selected. The treatment was given once a day, ten times of which made a session. After the treatment for 4 sessions, the clinical efficacy and Pittsburgh sleep quality index (PSQI) were compared between two groups.

RESULTSThe total effective rate was 92.6% (338/365) in the observation group, which was superior to 73.1% (242/331) in the control group (P < 0.05). The PSQI score was obviously decreased in two groups after the treatment (both P < 0.05), in which the decreasing in the observation group was superior to that in the control group (P < 0.05).

CONCLUSIONThe acupuncture prescription of regulating yin-yang and five viscera has better effect for intractable insomnia, which could be considered as a more optimal therapy.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Sleep ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Treatment Outcome ; Young Adult

BACKGROUNDEarly diagnosis and timely treatment are important for improving therapeutic efficiency of prostate cancer. DNA array is a new bio-technology for disease diagnosis. This study was conducted to diagnose prostate cancer with cDNA macroarray and analysis gene expression profiles of some selective genes in prostate cancer.

METHODSTotal RNA was isolated from patients with prostate cancer and from normal people, and poly (A) RNA was further purified. Then it was analyzed for differentially expressed genes in prostate cancer and normal prostate by cDNA macroarray system.

RESULTSThere were different expressions in the nine prostate-associated specific genes in prostate cancer as compared with normal prostate, in which, 7 were significantly upregulated and 2 were down-regulated.

CONCLUSIONAs a diagnostic approach at molecular level, the cDNA macroarray is an effectively diagnostic method for prostate cancer.

Gene Expression Profiling ; Genes, Tumor Suppressor ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; diagnosis ; genetics

OBJECTIVETo clinically study the antiviral effects of lamivudine and entecavir on patients with early-to-mid stage Hepatitis B related acute on chronic liver failure (HBV-ACLF). METHODS; A prospective, randomized, open and parallel controlled clinical trial was designed to observe the antiviral effects of nucleoside analogues on patients with early-to-mid stage HBV-ACLF. Three groups were set for controlled study, i. e. basic treatment group, lamivudine plus basic treatment group and entecavir plus basic treatment group.

RESULTSOne month after treatment, the improvement rates of lamivudine group and entecavir group were 58.85% and 59.15% respectively, significantly higher than that of basic treatment group which was 34.84% (Chi(2) = 9.8323, P = 0.043). By the end of six months, the cumulative survival rates of patients with the antiviral treatments, i.e., lamivudine, entecavir, were 65.8%, 60.1%, significantly higher than that (42%) without the antiviral treatment (P = 0.045, P = 0.04 respectively). The cumulative survival rate in patients with a MELD score < 30 was higher than that with a MELD score over 30 (Chi(2) = 3.920, P = 0.048). For the patients with pretreatment HBV DNA > or = 10(7), the cumulative survival rate in patients with entecavir treatments group was higher than that of patients in basic treatment group (Chi(2) = 5. 014 P= 0.025). According to the Ordinal Regression analysis, antiviral therapy by using either lamivudine or entecavia could significantly increase the improvement rate of patients with early-to-mid stage HBV-ACLF. But severe complications, including hepatorenal syndrome, electrolyte imbalance and hepatic encephalopathy, medical history of liver cirrhosis, and pretreatment HBV DNA > or = 10(7) had significant impacts on prognosis of this group patients.

CONCLUSIONSAntiviral therapy by using either lamivudine or entecavia could significantly increase the survival rate of patients with early-to-mid stage HBV-ACLF.

Anti-HIV Agents ; adverse effects ; therapeutic use ; Disease Susceptibility ; End Stage Liver Disease ; chemically induced ; Guanine ; adverse effects ; analogs & derivatives ; therapeutic use ; Humans ; Lamivudine ; adverse effects ; therapeutic use ; Prognosis

BACKGROUNDIt is accepted that inflammatory cytokines play a key role in the development of acute pancreatitis, so blocking the initiation of inflammatory reactions may alleviate pathological changes of acute pancreatitis. We studied the regulatory effect of arsenic trioxide (As(2)O(3)) on apoptosis and oncosis of pancreatic acinar cells in vitro and in vivo and its therapeutic effect on acute pancreatitis.

METHODSPancreatic acinar cells were isolated by collagenase digestion method. Apoptosis and oncosis of isolated pancreatic acinar cells were detected with Hoechst 33258 + PI or Annexin V + PI double fluorescent staining. Amylase and lactate dehydrogenase release were measured. Acute pancreatitis was induced in Wistar rats by intraperitoneal injections of caerulein, and apoptosis was detected with terminal dUTP nick-end labeling method. Tumor necorsis factor alpha (TNF-alpha) mRNA, myeloperoxidase, nuclear factor-kappaB and histological grading of pancreatic damage were measured.

RESULTSThere was an increased apoptosis but a decreased oncosis of pancreatic acinar cell after the treatment with As(2)O(3). The levels of lactate dehydrogenase and amylase release were markedly decreased in As(2)O(3) treated group. Myeloperoxidase content, TNF-alpha mRNA level, nuclear factor-kappaB activation and pathological score in As(2)O(3) treated group were significantly lower than in the untreated group.

CONCLUSIONSAs(2)O(3) can induce apoptosis and reduce oncosis of pancreatic acinar cell, thus resulting in reduced release of endocellular enzyme of acinar cells, reduced inflammatory cell infiltration and decreased the production of inflammatory cytokines, so that the outcome of alleviated pathological changes was finally achieved.

Amylases ; metabolism ; Animals ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Flow Cytometry ; In Situ Nick-End Labeling ; Inflammation ; drug therapy ; genetics ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; NF-kappa B ; metabolism ; Oxides ; pharmacology ; Pancreas ; drug effects ; metabolism ; pathology ; Pancreatitis ; drug therapy ; genetics ; metabolism ; Peroxidase ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; genetics

Objective To investigate the efficient method which can culture and induce embryonic stem cells to neurocyte in vitro. Methods Isolate the blastula of 3.5 d from BALB/c species mouse. Culture the cells from inner cell mass (ICM) which were isolated by mechanical method on the mouse embryonic fibroblaste cell (MEF) feeder layer or 0.1% gelatin coated dishes. The stem cells were identified by characterized morphology, alkaline phosphatase stain, differential potency in vivo and immunochemistry stain. The isolated cells were differentiated by serial induction method that mimicking the intrinsic developmental process of the neural system. Results The isolated cells were positive for alkaline phosphatatse and SSEA-1 (stage specific embryonic antigen 1). Moreover they were identified pluripotent by differentiation in vivo. Therefore the isolated cells presented the characters of ESCs. Then the isolated cells were able to differentiate into neurocytes in vitro. Conclusion Mouse embryonic stem cells isolation, culture and differentiation system has been established.