Objective To investigate the effects of anti-sense hTERT on the expression of VEGF and the receptor in SGC7901 cells, and study the influence in angiogenesis and progression in gastric carcinoma. Methods SGC7901 cell line was transfected by the recombinant virus containing sense and anti-sense hTERT cDNA. Then the expression of VEGF-C and Flt-4 was observed with RT-PCR, distribution of cell cycles was determined with flow cytometry. The expression of VEGF-C and Flt-4 protein was assessed with immunohistochemistry. Result The distribution of cell cycle of antisense hTERT transfected SGC7901 cells was changed, and the mRNA and protein expression of VEGF-C and Flt-4 was significantly down-regulated. Conclusion Anti-sense hTERT can act as an agent for inhibiting VEGF-C and Flt-4 mRNA and protein level, and it blocks tumor angiogenesis and lymphogenous metastasis.

BACKGROUND: Previous studies have demonstrated that LRP16 is an estrogen-responsive gene. Its expression level is strongly associated with the proliferation and invasive growth of human breast cancer cells.OBJECTIVE: To construct a LRP16 targeting vector and screen mouse embryonic stem cell clones with homolougous recombination of an inactive LRP16 gene.DESIGN: Constructing an inserting inactivation target by inserting SA-RIES-β geo expression cassette.SETTING: Bioregulatory Laboratory of the Third Medical Department of Kyushu University in Japan and Department of Molecular Biology, General Hospital of Chinese PLA.MATERIALS: The materials used here were mainly provided by the Bioregulatory Laboratory, the Third Medical Department of Kyushu University in Japan. The mouse genomic library in pBeloBAC11 Vector was purchased from lnvitrogen Corp. The competent TopF10 was purchased from Beijing Tiangen Biotech Corp. pcDNA3.1(+) vector was kept in our laboratory. Mouse ES cells were provided by Kyushu University.METHODS: The experiment was performed in Kyushu University and Department of Molecular Biology of PLA General Hospital from November 2004 to May 2005. Targeting sequence of LRP16 gene was obtained from 129 mouse genomic Bacterial Artificial Chromosomes library based on polymerase chain reaction (PCR) screening. The SA-RIES-β geo fragment was inserted within LRP16 fifth exon to inactivate LRP16. ES cells were screened with G418 and the homologously recombinant clone was identified by Southern blot analysis.MAIN OUTCOME MEASURES: Clones with homologous recombination.RESULTS: The LRP16 fragment including exon 5 to 11 was subcloned into the pBluescript SK Ⅱvector. Restriction map demonstrated that the SA-IRES-β geo fragment was correctly inserted into the LRP16 fifth exon. Southern blot results showed that there was an ES clone with targeting sequence homologously inserted.CONCLUSION: A LRP16 gene targeting vector is constructed and a homologous recombinant is obtained.

Objective To investigate the inhibitory effect of four and a half LIM protein 2(FHL2) on inhibitor of differentiation 1(Id1)-mediated suppression of transcriptional regulation activity and Id1-promoted invasive growth of human breast cancer cells MCF-7.Methods The effect of FHL2 on Id1-mediated transcriptional repression in MCF-7 cells was determined by cotransfection and relative biluciferase assay.The cell proliferation was determined by MTT assay,and the invasive capacity of MCF-7 cells was determined by Transwell assay.Results The transcriptional repression effect of Id1 on basic helix-loop-helix(bHLH) factor E47-mediated transcription activity in MCF-7 cells,and Id1-promoted proliferation and invasive growth of MCF-7 cells were significantly suppressed by FHL2.Conclusion FHL2 can inhibit the proliferation and invasive growth of human breast cancer cells via repressing Id1-mediated functional activity.The results provide a basis for further investigating the functional roles of FHL2-Id1 signal pathway in the carcinogenesis and development of human breast cancer.

OBJECTIVETo study the effects of Tongxinluo ultramicro-pulverization (TXLU) on experimental myocardial infarction and platelet aggregation of rats, investigate its mechanisms on ischemia heart disease and offer a reference to clinical usage.

METHODRats were separated randomly into 7 groups: sham, model, diltiazem (0.15 mg x kg(-1)), TXL(1.2 g x kg(-1)), TXLU (1.2, 0.6, 0.3 g x kg(-1)). The experimental myocardial infarction was induced with ligating the left anterior descending branch of the coronary of rats. The infarction size was determined after myocardium tissue was stained with 2,3,5-triphenyltetrazolium chloride (TTC). And the serum of rats was separated to analyze CK, LDH, SOD, MDA. Another 60 rats were separated randomly into 6 groups: control, aspirin (0.15 mg x kg(-1)), TXL (1.2 g x kg(-1)), TXLU (1.2 ,0.6,0.3 g x kg(-1)). The rat platelet aggregation was induced with adenosine diphosphate (ADP) and collagen to observe the inhibitory effects of TXLU.

RESULTTXLU could relieve the myocardial infarction size and weight stained with TTC significantly, the myocardial infarction size of the three groups of TXLU were (2.7 +/- 2.1)%, (3.4 +/- 1.2)%, (2.8 +/- 1.8)%, compared with model group (8.9 +/- 5.9)%, P < 0.05 or P < 0.01. The myocardial infarction weight of the three groups of TXLU were (8.4 +/- 3.5)%, (8.7 +/- 4.1)%, (9.7 +/- 4.1)%, compared with model group (l2.2 +/- 3.6)% P < 0.05 or P < 0. 01. And the content of MDA and the activities of CK and LDH in rats subjected with ligation of coronary artery were inhibited obviously too, compared with model group P < 0.05 or P < 0.01, then the activity of SOD increased. TXLU could inhibit the maximum percentage of rats platelet aggregation induced with ADP and collagen, the maximum percentage of platelet aggregation induced with ADP were (26.9 +/- 9.2)%, (24.4 +/- 13.4)%, (30.6 +/- 12.2)%, compared with control group (44.3 +/- 15. 7)% P < 0.05 or P < 0.01; The maximum percentage of platelet aggregation induced with collagen were (33.8 +/- 6.9)%, (32.1 +/- 8.3)%, (41.5 +/- 7.8)%, compared with control group (49.2 +/- 15.9)%, P < 0.05 or P < 0.01.

CONCLUSIONThe experiment results indicated that TXLU could protect myocardial tissue of rats from ischemic injury and the mechanism may be related with antioxidation and inhibiting platelet aggregation, and the results also suggested TXLU could lower clinical dosage.

Adenosine Diphosphate ; pharmacology ; Animals ; Aspirin ; pharmacology ; Diltiazem ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Male ; Medicine, Chinese Traditional ; methods ; Myocardial Infarction ; blood ; chemically induced ; drug therapy ; pathology ; Platelet Aggregation ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tetrazolium Salts ; pharmacology

BACKGROUND: The combination of polycaprolactone and hyaluronic acid creates an ideal environment for wound healing. Hyaluronic acid maintains a moist wound environment and accelerates the in-growth of granulation tissue. Polycaprolactone has excellent mechanical strength, limits inflammation and is biocompatible. This study evaluates the safety and efficacy of bio-conjugated polycaprolactone membranes (BPM) as a wound dressing. METHODS: 16 New Zealand white rabbits were sedated and local anaesthesia was administered. Two 3.0x3.0 cm full-thickness wounds were created on the dorsum of each rabbit, between the lowest rib and the pelvic bone. The wounds were dressed with either BPM (n=12) or Mepitel (n=12) (control), a polyamide-silicon wound dressing. These were evaluated macroscopically on the 7th, 14th, 21st, and 28th postoperative days for granulation, re-epithelialization, infection, and wound size, and histologically for epidermal and dermal regeneration. RESULTS: Both groups showed a comparable extent of granulation and re-epithelialization. No signs of infection were observed. There was no significant difference (P>0.05) in wound size between the two groups. BPM (n=6): 8.33 cm2, 4.90 cm2, 3.12 cm2, 1.84 cm2; Mepitel (n=6): 10.29 cm2, 5.53 cm2, 3.63 cm2, 2.02 cm2; at the 7th, 14th, 21st, and 28th postoperative days. The extents of epidermal and dermal regeneration were comparable between the two groups. CONCLUSIONS: BPM is comparable to Mepitel as a safe and efficacious wound dressing.
Bandages* ; Granulation Tissue ; Hyaluronic Acid ; Inflammation ; Membranes* ; Pelvic Bones ; Rabbits ; Re-Epithelialization ; Regeneration ; Ribs ; Skin ; Wound Healing ; Wounds and Injuries*

Gsalpha gene mutation has been discovered in some human tumors. In our previous studies, three novel deletants of Gsalpha gene, Gsalpha L-1(500 bp), Gsalpha L-2(300 bp), and Gsalpha L-3(200 bp), and wild type Gsalpha-4(1 200 bp) were found in human leukemia cell lines and detected in leukemic cells from patients with acute leukemia. To investigate the construction, function and biological significance of the deletants, the plasmids of Gsalpha L-1, Gsalpha L-2 and wild Gsalpha-4 were transformed into E. coli DH5, amplified by PCR, and cloned in expression vector pET22b(+), and then transformed into E. coli, respectively. As a result, higher levels of expression of three recombinants were obtained in form of inclusion bodies. The results suggested that these Gsalpha isoforms have an open reading frame of gene and can be expressed in vitro. The data lay a foundation to study the relation of Gsalpha gene to leukemogenesis.

Objective:To explore the changes of sympathetic and parasympathetic activity in children with chest pain and/or chest distress of unknown cause based on their plasma catecholamine and heart rate variability indexes, and to analyze the predictive value of each index in diagnosing pediatric sympathetic excitation-related chest pain and/or chest distress by used receiver operating characteristics (ROC) curve analysis.Methods:Fifty-seven children who complained of unexplained chest pain and/or chest distress and had no organic diseases according to routine examinations in the Chengdu Women′s and Children′s Hospital from June 2017 to June 2019 were enrolled in the study group.There were 22 males and 35 females, aged 5-15 years old[(8.40±0.35) years old]. Meanwhile, 54 healthy children in the same period were enrolled in the healthy control group, including 21 males and 33 females, aged 5-15 years old (8.87±0.36) years old]. The disease history enquiring, physical examination, chest X-ray, 12-lead electrocardiogram, echocardiogram, blood routine test and biochemical test excluded the organic heart, lung, digestive tract and chest wall diseases in both groups.Their blood samples were collected to detect catecholamine and they were monitored by dynamic electrocardiogram.SPSS 25.0 software was used for statistical analysis and ROC curve analysis.Results:The study group exhibited higher dopamine, adrenaline and norepinephrine levels than those in the healthy control group[(0.83±0.04) nmol/L vs.(0.54±0.03) nmol/L, (0.76±0.04) nmol/L vs.(0.56±0.03) nmol/L and(3.59±0.18) nmol/L vs.(2.51±0.15) nmol/L], and the differences were statistically significant( t=4.906, 3.611, 4.596, all P<0.01). The levels of standard deviation of NN intervals, standard deviation of all mean 5-minutes NN intervals, standard deviation of all NN intervals for all 5-minute segments of 24 hours, root mean squared successive difference and proportion of NN 50 in the total number of NN intervals in the study group were lower than those in the healthy control group [(110.49±2.81) ms vs.(132.13±2.55) ms, (86.37±3.26) ms vs.(118.96±2.00) ms, (33.46±2.21) ms vs.(68.91±1.29) ms, (37.63±1.22) ms vs.(48.93±1.75) ms and(17.37±1.45)% vs.(22.22±1.61)%], and the differences were statistically significant( t=-5.710, -8.419, -13.862, -5.354, -2.245, all P<0.05). The area under the curve of plasma dopamine, adrenaline and norepinephrine in the diagnosis of pediatric sympathetic excitation-related chest pain and/or chest distress were 0.753, 0.689 and 0.746, respectively, higher than that of all heart rate variability indexes. Conclusions:Children′s unexplained chest pain and/or chest distress is highly related with autonomic nervous dysfunction (increased sympathetic activity and decreased parasympathetic activity). The level of plasma catecholamine can be used to reflect the changes of sympathetic activity in these children and diagnose sympathetic excitation-related chest pain and/or chest distress.Psychological counseling and relaxation therapy are supposed to be effective in redu-cing sympathetic activity, improving subjective symptoms and elevating their quality of life.