[Antitumor and antimetastatic effect of antimicrobial peptide conjugated with tumor homing peptide TMTP1 on the transplanted prostate cancer and gastric cancer in nude mice].

OBJECTIVE: Due to their lower risk for induction of resistance, antimicrobial peptides with selective anticancer effect could be developed into a new generation of anticancer drugs. We conjugated an antimicrobial peptide with tumor-targeting peptides (TMTP1) to explore whether it has inhibiting effect on the progression and metastasis of transplanted prostate cancer and gastric cancer in nude mice. METHODS: Subcutaneously transplanted human prostate cancer and orthotopically transplanted human gastric cancer in nude mice were prepared. 50 µmol/L PBS (control group), 50 µmol/L TMTP1 (TMTP1 group) or 50 µmol/L TMTP1-GG-D(KLAKLAK)(2) (treatment group) were injected i.p. to the three groups of nude mice, respectively. The binding ability of the novel fusion polypeptide TMTP1-GG-D(KLAKLAK)(2) to the tumors and its antitumor effect were assessed by measurement of tumor volume, histopathological examination of the tumor tissues, testing apoptosis index of tumor cells with TUNEL staining, and survival curve plotting of the mice. RESULTS: The median survival time of subcutaneous prostate cancer-bearing mice was 50 days in the control group, 55 days in the TMTP1 group, and 70 days in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.05). The median survival time of the subcutaneous gastric cancer-bearing mice was 25 days in the control group, 30 days in the TMTP1 group, and 45 days in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.01). The tumor volume in the subcutaneous prostate cancer-bearing mice was (2.5 ± 0.3)cm(3) in the control group, (1.8 ± 0.2) cm(3) in the TMTP1 group, and (0.3 ± 0.1)cm(3) in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.01). The tumor volume of the subcutaneous gastric cancer-bearing mice was (3.8 ± 0.4) cm(3) in the control group, (3.2 ± 0.2)cm(3) in the TMTP1 group, and (0.4 ± 0.1) cm(3) in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.01). Large tumors were observed in the stomach of the orthotopic gastric cancer-bearing mice of the control and TMTP1 groups. The tumor volume of the TMTP1-GG-D(KLAKLAK)(2) group was obviously reduced. White metastases in the liver, spleen and abdominal wall were observed in the control and TMTP1 groups (P < 0.01). TUNEL staining revealed that the apoptosis index of the control group was (31.9 ± 1.5)%, TMTP1 group (37.2 ± 2.3)% and TMTP1-GG-D(KLAKLAK)(2) group (69.7 ± 2.1)% (P < 0.01). CONCLUSIONS: The results of our study demonstrate that the novel fusion peptide of antimicriobial peptide conjugated with TMTP1 can effectively inhibit tumor progression and metastasis, therefore, is promising to be a novel effective anticancer drug.

Kallikrein gene transfer induces angiogenesis and further improves regional cerebral blood flow in the early period after cerebral ischemia/reperfusion in rats.

AIMS: The aims of this study were to find out whether kallikrein could induce angiogenesis and affect the cerebral blood flow (rCBF) in the early period after cerebral ischemia/reperfusion (CI/R). METHODS: The adenovirus carried human tissue kallikrein (HTK) gene was administrated into the periinfarction region after CI/R. At 12, 24, and 72 h after treatments, neurological deficits were evaluated; expression of HTK and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry staining; the infarction volume was measured; and rCBF was examined by( 14) C-iodoantipyrine microtracing technique. RESULTS: The expression of VEGF was enhanced significantly in pAdCMV-HTK group than controls over all time points (P < 0.05). Furthermore, the rCBF in pAdCMV-HTK group increased markedly than controls at 24 and 72 h after treatment (P < 0.05), and the improved neurological deficit was accompanied by reduced infarction volume in pAdCMV-HTK group 24 and 72 h posttreatment. CONCLUSION: In the early period after CI/R, kallikrein could induce the angiogenesis and improve rCBF in periinfarction region, and further reduce the infarction volume and improve the neurological deficits.

Clinical observation on the acupuncture treatment in patients with urinary retention after radical hysterectomy.

OBJECTIVE: To observe the acupuncture therapy effect on the urinary retention after radical: hysterectomy. METHODS METHODS: Eighty cases of urinary retention after radical hysterectomy were randomly assigned to: the treatment group and control group according to the random number table method, 40 cases in each group. From the 15th day post operation, the patients in the two groups started to be treated and 7 days as a course with 5 days treatment and 2 days interval. The treatment group applied acupuncture with modalities of common needling on Zusanli (ST36) bilaterally, electroacupuncture on Sanyinjiao (SP6), Shuidao (ST28), and Scalp Reproduction Area bilaterally, moxibustion on Shenque (CV8). The control group applied acupoint injection with vitamin B B12 12, and Sanyinjiao and Zusanli were selected. Take turns on both sides. The courses for the recovery, of bladder function and residual urine volume for those who had voluntary micturition more than 200 mL after the first and second course of treatment were compared between the two groups. RESULTS: Within 1 course and 2: courses of treatment, the patients with bladder function recovery in the treatment group were 21 (21/40) and 36 (36/40), and those in the control group were 12 (12/40) and 29 (29/40), both with a significant difference (P<0.05). After the first course and second course, residual urine volume for those who had voluntary micturition more than 200 mL in the treatment group was 91.7 ± 17.5 mL and 93.5 ± 15.5 mL, in the control group 102.4 ± 13.7 mL and 102.5 ± 15.7 mL, both with a significant difference (P<0.05). CONCLUSIONS: Combination of acupuncture: modalities was better than acupoint injection for the recovery of bladder function in urinary retention after radical hysterectomy. It would shorten the course of treatment and get a better recovery, decrease urinary retention cases of refractoriness, and extending the treatment could raise the healing rate of urinary retention.

[Clinical management of mandibular incisors with multiple root canals using dental operating microscope].

OBJECTIVE: To evaluate the role of dental operating microscope in clinical treatment of lower incisors with multiple root canals. METHODS: 143 mandibular incisors in 128 patients were treated endodontically. Two kinds of preoperative radiographs were taken for each tooth, using straight projection and eccentric projection. The root canal morphology was recorded according to Vertucci's classification. Under dental operating microscope, the teeth with multiple root canals were shaped using Ni-Ti rotary instruments Hero 642, cleaned using sodium hypochlorite, and obturated with vertical condensation technique. The following information was recorded: The number of teeth that were found to have multiple canals in two kinds of preoperative radiographs, and when using and without using microscope. The efficiency of preparation and obturation was analyzed with radiographs before, during and after operation. RESULTS: The mandibular central incisor with one canal was 73.53% and multiple canals was 26.47% in treated teeth. The mandibular lateral incisor with one canal was 70.67% and multiple canals was 29.33% in treated teeth. By eccentric projection radiograph and treatment using microscope, more teeth with multiple canals were found. No complication was found during root canal preparation. 134 teeth were well filled and 9 showed slight over-filling. CONCLUSION: With dental operating microscope, the mandibular incisors with multiple root canals could be treated well in combination with rotary instrumentation and vertical condensation technique.

[Role of human telomerase reverse transcriptase in apoptosis of human umbilical vein endothelial cells induced by trichostatin A].

OBJECTIVE: The aim of this study was designed to investigate the effect of TSA on human umbilical vein endothelial cells and to reveal its possible mechanisms and relationship between apoptosis and activity of telomerase reverse transcriptase. METHODS: sulforhodamine B method was employed to determine the growth rate of umbilical vein endothelial cells. The cell apoptotic rate was measured by flow cytometry (FCM). The hTERT and p21(Waf1) mRNA expression before and after TSA treatment were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The quantitative of hTERT protein expression in cells were detected by flow cytometry. After transfection, the cell telomerase activity was detected by PCR and telomeric repeat amplification protocol assay (PCR-TRAP-ELISA) and early apoptosis was measured by Annexin V/PI stain and flow cytometry. RESULTS: After being treated with TSA, the proliferation of umbilical vein endothelial cells was inhibited. Slight apoptosis and cell cycle arrest were detected. However, the same concentration of TSA induced serious apoptosis in HeLa cells. Up-regulation of hTERT mRNA expression was observed within 48 h after TSA treatment, but the change of p21(Waf1) expression was not significant. The umbilical vein endothelial cells hTERT protein expression level was increased within 24 h. After transfection of the dominant negative, wild type and control hTERT plasmid, a significant difference of telomerase activity in these cells was observed by PCR-TRAP-ELISA assay. WT-hTERT-transfected cells were more resistant to apoptosis induced by trichostatin A. CONCLUSION: Human umbilical vein endothelial cells could be resistant to apoptosis induced by high concentrate TSA, and hTERT might play an important role in this process.