Objective To investigate the expressions of COX-2 and p53 in human colorectal carcinoma and their correlations with clinicopathological features of colorectal carcinoma. Methods The expressions of COX-2 and p53 were determined by immunohistochemical staining in 72 surgical specimens of colorectal carcinoma and 21 normal mucosal tissues. Results The positive rates of COX-2 and p53 in colorectal carcinoma were 70.8 % and 62.5 %, respectively. The differences between carcinoma group and normal tissues group were statistically significant(P

Objective To observe the effect of Glutsmine on intestinal barrier function after traumatic brain injury in rats. Methods A total of 54 adult male Wistar rats were divided randomly into 3 groups, ie, normal group (Group N, 6 rats), TBI group (Group T, 24 rats) and Giutamine intervention group (Group G, 24 rats). Group T and Group G were subdivided into 4 groups according to detection time at days 1,3, 5 and 7 respectively. Meanwhile, 6 rats were enlisted in each group. The intestinal mucosa structure was detected by histopathological examination and electron microscopy. Apoptosis was detected by in situ immunohistochemical staining (TUNEL). Results Glutamine could relieve the pathological lesion of gut mucosa and decrease intestinal mucosa cell apoptosis after traumatic brain injury. Conclusion Glutamine can protect intestinal mucosa barrier function following traumatic brain injury.

BACKGROUND:Specific down-regulation of interleukin 1 beta (IL-1β) may al eviate the pain behaviors effectively after peripheral nervous injury. Compared with smal interference RNA (siRNA), short hairpin RNA (shRNA) could inhibit the expression of target gene more stably and efficiently. However, simple shRNA could not enter target cel s to down-regulate target gene efficiently. Adenovirus vectors have wide host range, high infection efficiency and stable expression in host cel s.
OBJECTIVE:To construct recombinant adenovirus vector expressing shRNA targeting IL-1βgene and detect its effect on the expression of target gene.
METHODS:Three siRNAs were designed on the basis of the nucleotide sequence of IL-1βobtained from NCBI and then three shRNAs (shRNA1, shRNA2 and shRNA3) were synthesized. The annealed shRNA product and adenovirus vector pHBAd/U6/GFP digested by BamH I and EcoR I were connected to construct the recombinant adenovirus vector shuttle plasmid expressing shRNA targeting IL-1β. After sequencing, HEK 293 cel s were co-transfected by the shuttle plasmid and skeleton vector, and three recombinant adenovirus vector expressing shRNA targeting IL-1β(rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3) were packaged and amplified. Rats H9C2 cel s were infected by recombinant adenovirus vector expressing shRNA targeting IL-1βand fluorescence microscope was used to observe the infection efficiency. The effect of recombinant adenovirus vector expressing shRNA targeting IL-1βon the expression of target gene was detected by western blot assay.
RESULTS AND CONCLUSION:The sequencing results showed that the sequences of three shRNAs adenovirus vector shuttle plasmid were consistent with the sequences of three designed shRNAs. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 were constructed successful y. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 could down-regulate the expression of IL-1βin rat H9C2 cel s and the down-regulation effect of rAd/shRNA2 was the most significant.

Objective To evaluate the role of interleukin?1β( IL?1β) in activation of N?methyl?D?aspartate ( NMDA) receptors in spinal dorsal horns of rats with neuropathic pain. Methods One hundred twenty?eight pathogen?free healthy adult male Sprague?Dawley rats, weighing 200-250 g, were divided into 4 groups (n=32 each) using a random number table: control group (group C), recombinant adenovirus vector group ( group rAd) , recombinant adenovirus vector shRNA group ( group rAd∕shRNA) , and recom?binant adenovirus vector shRNA plus recombinant adenovirus vector IL?1β group ( group rAd∕shRNA+rAd∕IL?1β) , receiving intrathecally injected equal volume of 0.9% sodium chloride solution, recombinant ade?novirus vector, recombinant adenovirus vector shRNA, and recombinant adenovirus vector shRNA plus re?combinant adenovirus vector IL?1β5 μl, respectively. Chronic compression of dorsal root ganglia was per?formed on 8th day after intrathecal injection. The thermal paw withdrawal latency ( TWL) was measured at 1 day before intrathecal injection ( T0 ) and at 1, 2, 3 and 5 weeks after intrathecal injection ( T1-4 ) . After measurement of the pain threshold at each time point, 8 rats selected randomly were anesthetized and sacri?ficed, and the lumbar enlargement segments of the spinal cord were harvested for determination of the ex?pression of IL?1β and phosphorylated NMDA receptor NR1 subunits at serine 896 ( pNR1S896) in spinal dorsal horns on the injured side by immunohistochemistry and Western blot. Results Compared with group C, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA (P<0.05), and no significant change was found in the parameters mentioned above at each time point in group rAd and group rAd∕shRNA+rAd∕IL?1β( P>0.05) . Compared with group rAd, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA ( P<0.05) , and no significant change was found in the parameters mentioned above at each time point in group rAd∕shRNA+rAd∕IL?1β (P>0.05). Compared with group rAd∕shRNA, the TWL was significantly short?ened, and the expression of IL?1β and pNR1S896 in spinal dorsal horns was significantly up?regulated at T2-4 in group rAd∕shRNA+rAd∕IL?1β ( P<0.05) . Conclusion IL?1β is involved in the activation of NM?DA receptors in spinal dorsal horns of rats with reuropathic pain.

ObjectiveTo investigate the changes in the expression of T-cell death-associated gene 8(TD- AG8) in spinal cord in rats with bone cancer pain.MethodsTwo hundred and twenty-four female rats weighting 150-180 g were randomly divided into 3 groups:normal control group(group Ⅰ,n ＝ 64),normal saline group (group Ⅱ,n ＝ 64),bone cancer pain group(group Ⅲ],n ＝ 96).Bone cancer pain was induced by inoculating Walker256 mammary gland carcinoma cells into the tibia medullary cavity.Mechanical withdrawl threshold(MWT)was measured at 1 d before(baseline)and 1,3,6,9,12,15 and 18 d after inoculation.Sixteen rats were sacrificed at 1 day before(baseline)and 6,9,12,15 and 18 d after inoculation in group Ⅲ and 18 d after inoculation in groups Ⅰ and Ⅱ.The L4-6 spinal cord were removed,and the number of TDAG8 positive cell was counted,and the expression of TDAG8 mRNA was measured by RT-PCR.ResultsCompared with baseline value and group Ⅰ,MWT was decreased,and the number of TDAG8 positive cells and the expression of TDAG8 mRNA in spinal cord were increased at 6-18 d after inoculation in group Ⅲ ( P < 0.01 ).ConclusionThe expression of TDAG8 in spinal cord is up-regulated in rats with bone cancer pain,which may be involved in the mechanism of the development of bone cancer pain.

Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38％ of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95％,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.

OBJECTIVETo observe the inhibition of NK4 protein in the proliferation of human Raji lymphoma xenografts in nude mice, and to explore its molecular mechanism.

METHODSModels of human Raji lymphoma xenograft transfected with HGF gene were established by subcutaneous inoculation in nude mice. After establishment of the models, the mice received continuous NK4 protein via tail vein for 4 weeks, and the weight and tumor growth were monitored every week. After 8 weeks, the expression of HGF mRNA and c-Met mRNA of tumor tissues was measured by real-time fluorescent quantitation PCR. The apoptotic index (AI) and microvessel density (MVD) were evaluated by terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) and immunohistochemistry, respectively.

RESULTSThe models of human Raji lymphoma xenograft were successfully established. Although the animal weights of all groups declined, especially in the groups with NK4 protein injection, there was no statistical significance (P > 0.05). The tumor volume in HGF gene transfected group was larger than those of the control groups (P < 0.01), and there was no statistical significance among the control groups (P > 0.05). However, the tumor volume of the NK4 protein injection group decreased significantly (P < 0.01). Expression of HGF mRNA and c-Met mRNA in HGF gene transfected group increased significantly after injection of NK4 protein (P < 0.01). AI in HGF gene transfected group (33.5% ± 12.3%) was significantly lower than that of control groups (89.1% ± 22.3% vs. 81.9% ± 27.0%, P < 0.05), but became significantly higher (119.1% ± 18.9%) after NK4 protein injection (P < 0.01). MVD in HGF gene transfected group (28.5 ± 2.0) was higher than that of control groups (12.2 ± 1.4, 13.8 ± 1.3, P < 0.01), although declined (15.5 ± 2.5) after NK4 protein injection (P < 0.01).

CONCLUSIONSNK4 protein suppresses significantly the growth of human Raji lymphoma xenografts transfected with HGF gene. The pathogenesis may be involved in promoting tumor cell apoptosis and restraining tumor angiogenesis through competitive interrupting HGF/Met signal pathway.

Animals ; Apoptosis ; Hepatocyte Growth Factor ; genetics ; metabolism ; Heterografts ; Humans ; Lymphoma ; genetics ; metabolism ; therapy ; Mice ; Mice, Nude ; Microvessels ; pathology ; Neovascularization, Pathologic ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; T-Box Domain Proteins ; administration & dosage ; Transfection ; Transplantation, Heterologous

Objective To evaluate the efficacy and safety of motherwort (herbs leonuri/leonurus heterophyllus sweet) injection for preventing postpartum hemorrhage after caesarian section. Methods The prospective study was designed as a randomized and single blind multi-center research matched with positive agent as controls from Apt 2007 to Aug 2007. 440 women underwent caesarian section (CS) indicated by obstetric factors were enrolled from 15 teaching hospitals in China and assigned into three groups: group of motherwort: 147 cases were administered by motherwort 40 rag uterine injection during CS and 20 mg intramuscular injection per 12 hours 3 times after CS; group of motherwort + oxytocin : 144 cases were administered by motherwort 40 mg and oxytocin 10 U uterine injection during CS and motherwort 20 mg intramuscular injection per 12 hours 3 times after CS and group of oxytocin: 149 cases were administered by oxytocin 10 U uterine injection and oxytocin 10 U + 5% glucose 500 nd intravenously injection during operation and oxytocin 10 U intramuscular injection per 12 hours 3 times after CS. The following clinical parameter were collected and analyzed: (1) The amount of blood loss during operation, at 2, 6, 12, 24, 48 hours after operation. (2) The total amount of blood loss in 24 hours after CS and the incidence of postpartum hemorrhage. (3) The change of level of hemoglobin (Hb) and counting of red blood cell ( RBC ) from prepartum to postpartum. (4) Adverse reaction. Results (1) The mean amount of blood loss during operation were (368±258) ml in group of motherwort, (255±114) mi in group of motherwort + oxytocinand (269±141 ) ml in group of oxytocin, which exhibited significant difference among three groups ( P<0.01 ). Meanwhile, no statistical different amount of blood loss among three groups were observed at 2,6,12, 24, 48 hours after CS. (2)The amount of blood loss of postpartum at 24 hours were (480±276)ml ingroup of motherwort, (361±179) ml in group of motherwort + oxytocin, (381±179) nd in group of oxytocin, which showed significant difference among 3 groups(P <0.01 ). (3) The incidence of postpartum hemorrhage were 32.0% (47/147) in group of motherwort, 11.1% (16/144) in group of motherwort + oxytocin, and 18.8% in (28/149) in group of oxytocin. When comparing the lowest rate of postpartum blood loss in group of motherwort + oxytocin and the highest rate in group of motherwort, it displayed statistical difference (P<0.01). (4) The decreased level of RBC and Hb were shown that RBC(0.3±0.5)×10<'12<‘/L and Hb(9±13)g/L in group of motherwort, RBC (0.2±0.4)×10<'12/L and Hb ( 6±10) g/Lin group of motherwort + oxytocin and RBC (0.2±0.4)×10<'12/L and Hb(7±30) g/L in group of oxytocinrespectively. However, the comparison of different value of RBC and lib in group of oxytocin and motherwort +oxytocin showed significant difference (P<0.05 ). (5) Two cases with allery reaction was observed.Conclusion It is safe and efficacious that combined use of motherwort injection and oxytocin was to preventpostpartum hemorrhage during or after caesarian section.

【Objective】 To share the technical key points and experience of transvesical robot-assisted radical prostatectomy (TvRARP). 【Methods】 The clinical data of 13 patients with prostate cancer (PCa) receiving TvRARP during Nov.2021 and May 2022 were collected. The operation time, estimated blood loss, blood transfusion rate, catheter removal time, postoperative length of hospital stay, immediate urinary continence rate, postoperative IIEF-5 score and perioperative complications were evaluated. 【Results】 The operation time was (142±39) min, estimated intraoperative blood loss was (76±40) mL, and no transfusion was needed. The median postoperative IIEF-5 score was 16 (12-22), hospital stay 3 (2-5)days, and catheter removal time 7(5-14)days. Of all 13 patients, 12(92.3%) achieved immediate urinary continence at the removal of catheter. There were no postoperative complications of Clavien Ⅲ and above. Clavien Ⅰ-Ⅱ complications were observed in 4 patients (30.8%). 【Conclusion】 TvRARP is feasible and safe for selected patients with clinically localized PCa, which can ensure promising postoperative urinary continence and preserve erectile functional.