Objective To determine whether cultured neural stem cell expresses morphogen molecule sonic hedgehog′s functional receptor patched. Methods Cultured neural stem cell clones were subjected to RT\|PCR after passaged several times in vitro ,the amplification production was sequenced and labeled by digoxingemin and in situ hybridization technique was carried out to detect the cryosection of the neural stem cell clones. Results Most cells in the stem cell clones were positive for sonic hedgehog functional receptor patched,no significant difference was found among the positive cells and the center and peripheral of the stem cell clones.Conclusion\ The sonic hedgehog signal transduction may have important role in the proliferation and differentiation process of neural stem cell.\;[

Objective: This present study is to establish a rat model of intravenous heroin self-administration and observe the effect of electro-acupuncture on the heroin-seeking behavior of the heroin self-administrating rat. Methods: Six Male Wistar rats were surgically implanted with an intra-jugular intravenous catheters under sodium pentobarbital anesthesia,following a minimum recovery of 7 days, the rats were trained by nose-poke response with one daily 4-hour session for 14 consecutive days to self administer heroin (50 μg /kg / per injection), the session ended after 25 infusions were earned or 4 hours had passed, whichever came first, the reinforcement schedule was fixed ratio 1 (FR1) that the rat received a heroin injection for each nose poke response. And then the rats were given electro-acupuncture treatment daily 30min prior to each session and then were trained with one daily 4-h session for 7 consecutive days. Results: Stable intravenous heroin self-administration behavior of rats was established after 7-8 days' drug training, electro-acupuncture given to the stable heroin self-administrating rat 30 min prior to each session could reduce the drug self-administration behavior, namely the total heroin intake in each session was reduced, especially the 1st hour heroin intake, the initiation of first nose-poke response in each session was relayed, and the haste heroin-seeking behavior also became slower. Conclusions: The above results suggested that the electro-acupuncture had better detoxification effect.

OBJECTIVEThe purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.

METHODSA pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.

RESULTSThe results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.

CONCLUSIONThe RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

Caliciviridae Infections ; diagnosis ; virology ; DNA Primers ; genetics ; Gastroenteritis ; diagnosis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods

AMP-Activated Protein Kinases ; metabolism ; Animals ; Male ; Mice ; Mice, Inbred C57BL ; Muscle, Skeletal ; metabolism ; Phosphorylation ; Physical Conditioning, Animal

OBJECTIVETo investigate the role of endoplasmic reticulum stress in the apoptosis of testicular germ cells in hyperlipidemic rats.

METHODSWe randomly assigned 42 four-week-old male Wistar rats into a normal control group (n = 12) and a high-fat group (n = 30) to be fed on a normal diet and a high-fat diet, respectively, for 10 weeks. Then we measured the concentrations of triglyceride (TG) and total cholesterol (TC) in the serum using an automatic biochemistry analyzer, detected the apoptosis of testicular germ cells by TUNEL staining, and determined the protein and mRNA expressions of GRP78 and. caspase-12 in the testis tissue by immunohistochemistry and RT-PCR, respectively.

RESULTSThe concentrations of TG and TC were significantly increased in the animals of the high-fat group ([3.00 ± 0.92] and [3.04 ± 0.39] mmol/L) as compared with the control rats ([1.43 ± 0.41] and [1.55 ± 0.23] mmol/L) (P < 0.01), and so was the apoptosis index of the testicular germ cells ([37.17 ± 2.74]% vs [5.16 ± 0.81]%, P < 0.01). The high-fat group, in comparison with the control, also showed remarkably upregulated protein and mRNA expressions of GRP78 (0.32 ± 0.03 and 0.86 ± 0.05 vs 0.19 ± 0.01 and 0.37 ± 0.03, P < 0.01) and caspase-12 (0.34 ± 0.02 and 0.87 ± 0.01 vs 0.12 ± 0.01 and 0.34 ± 0.03, P < 0.01) in the testis tissue.

CONCLUSIONThe apoptosis of testicular germ cells is increased in hyperlipidemic rats, which may be attributed to endoplasmic reticulum stress.

Animals ; Apoptosis ; physiology ; Caspase 12 ; metabolism ; Cholesterol ; blood ; Diet, High-Fat ; adverse effects ; Endoplasmic Reticulum Stress ; physiology ; Heat-Shock Proteins ; metabolism ; In Situ Nick-End Labeling ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Spermatozoa ; pathology ; Staining and Labeling ; Testis ; metabolism ; Transcriptional Activation ; Triglycerides ; blood ; Up-Regulation

In this study, a composite cell model for evaluation of idiosyncratic drug-induced liver injury (IDILI) was established in vitro from the perspective of immune inflammation. And this model was used to evaluate the risk of IDILI for 2,3,5,4'-tetrahydroxy-cis-stilbene-2-O-β-glucoside (Cis-SG) and 2,3,5,4'-tetrahydroxy-trans-stilbene-2-O-β-glucoside (Trans-SG). To determine the low, medium, and high dosage of Cis-SG and Trans-SG, CellTiter-Glo^® 3D Cell Viability Assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of HepG2 cells in three dimensional (3D) culture, and MTT assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of THP-1 derived macrophages. THP-1 derived macrophages were incubated by Cis-SG and Trans-SG directly or supernatants from HepG2 cells incubated with them. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) in the supernatants of the THP-1 derived macrophages. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression of apoptosis-associated speck-like protein (ASC), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (caspase-1), and IL-1β in THP-1 derived macrophages. The results showed that there was no effect on the secretion of IL-1β in THP-1 derived macrophages incubated by Cis-SG and Trans-SG directly. However, the secretion of IL-1β, the protein and mRNA expression of ASC, NLRP3, caspase-1, and IL-1β significantly increased in THP-1 derived macrophages incubated by supernatants from HepG2 cells incubated with 1, 5, and 25 μmol·L^-1 Cis-SG or 25 μmol·L^-1 Trans-SG. In summary, the composite cell model for evaluation of IDILI established in vitro has been successfully applied in testing Cis-SG and Trans-SG. This composite cell model is helpful to evaluate and screen drugs with IDILI risk in vitro preliminarily, which provides methods for predicting and solving the idiosyncratic liver toxicity of drugs.

Aged ; Drug-Related Side Effects and Adverse Reactions ; blood ; therapy ; Emergency Treatment ; Female ; Hemoperfusion ; Humans ; Hypnotics and Sedatives ; poisoning ; Male ; Middle Aged ; Serum ; chemistry

OBJECTIVETo observe the clinical effect of Guilu Erxian Glue Cataplasm (GEGC) on carcinoma of the large intestine patients with myelosuppression after chemotherapy, and further to confirm its efficiency and safety.

METHODSTotally 60 patients with carcinoma of the large intestine were randomly assigned to two groups. Meanwhile, they all accepted FOLFIRI chemotherapy. Patients in the treatment group were additionally applied at Shenque (RN8), exchanging once per every other day, for 14 successive days. Patients in the control group took placebos with the same dose and dosage as the treatment group. The blood cell counts (WBC, NE, and PLT) were detected before chemotherapy, at day 7, 10, and 14. The TCM symptoms integrals, Karnofsky performance score (KPS), liver and kidney functions were observed before chemotherapy, at day 7 and day 14. Adverse skin reactions were observed each day. And the usage of hematopoietic growth factors was recorded.

RESULTS(1) The KPS score at day 7 was more stable in the treatment group than in the control group; the WBC and NE counts in the peripheral blood at day 14 were higher in the treatment group than in the control group; and TCM symptoms integrals at day 14 was lower in the treatment group than in the control group, all with statistical difference (P < 0.05). (2) Compared with the control group, the PLT count was higher in the treatment group than in the control group, the usage of rhG-CSF and antibiotics was less in the treatment group than in the control group, all with no statistical difference (P > 0.05). (3) No obvious adverse reactions such as liver injury, renal injury, or skin allergy were observed.

CONCLUSIONSAdjuvant treatment of GEGC could improve carcinoma of the large intestine patients with myelosuppression to some extent. No relevant adverse reactions were found.

Adjuvants, Immunologic ; therapeutic use ; Adult ; Aged ; Bone Marrow Diseases ; chemically induced ; drug therapy ; Colorectal Neoplasms ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo explore the effect of a self-made guiding needle of steel wire in guiding the wire through the tibial tunnel for the treatment of avulsion fractures of tibial posterior cruciate ligament with open reduction and wire fixation.

METHODSFrom February 2011 to June 2014, a total of 22 patients with avulsion fractures of tibial posterior cruciate ligament underwent surgical treatments were analyzed, including 14 males and 8 females with an average age of 35.6 years old (ranged, 17 to 63 years old). According to Meyers classification, 9 patients were classified as type II, 13 patients were classified as type III. All the patients underwent open reduction and wire fixation with medial knee "L" shape approach. A wire guiding needle was used to guide the wire through the tibial tunnel during operation.

RESULTSWith the assistance of wire guidance needles, wires passed through the tibial tunnel rapidly during the operation in all the 22 patients. All the patients were followed up, X-ray imagings 6 months after operation showed the fractures healed well. The average follow-up time in all patients was 6 months (ranged, 6 to 12 months). The averaged Lysholm knee score in 22 knee was 92.7 +/- 3.4. All patients' posterior drawer test were negative.

CONCLUSIONSelf-made wire guiding needle can simplify the operation procedures in which the wires pass through the tibial tunnel, shorten the operation time, reduce the surgical trauma and complications, and be worthy of clinical application.

Adolescent ; Adult ; Bone Wires ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Posterior Cruciate Ligament ; injuries ; surgery ; Tibia ; injuries ; surgery ; Tibial Fractures ; surgery ; Young Adult

Objective To design a new method to verify the position of multileaf collimator(MLC)leaf using a two-dimensional ion chamber array(2D-array).Methods 2D-array of PTW T10018 Seven29~(TM) was used to calibrate the accuracy of MLC leaf position of Elekta Precise accelerator.The edge function of the leaf position of MLC was measured and used as the reference value.The precision of MLC leaf was then evaluated through comparing the measured and reference values.Results The accuracy of MLC leaf position was found within?0.1 mm.Conclusion This method of verifying the accuracy of multileaf collimator leaf position is easy,simple and reliable