OBJECTIVETo investigate the effect of MTS1 gene beta promoter transcriptional activation in T-cell acute lymphoblastic leukemia (T-ALL) cell lines and identify the fragment with transcriptional activation.

METHODSSeven pGL3 recombinant plasmids with the same 3'-end transcriptional start site but the different 5'sequences were constructed by gene recombinant technique and transfected into Jurkat cell line which is biallelic deletion of MTS1 gene by transient transfection. Luciferase report gene was detected to observe beta promoter transcriptional activation.

RESULTSSeven pGL3 recombinant plasmids containing different fragments of beta promoter were obtained, all of them showed transcriptional activation in Jurkat cell line. Among them, the 0.38 kb fragment cut by SacII-SacI is fundamental in transcription.

CONCLUSIONMTS1 gene beta promoter can be activated in Jurkat cell line.

Genes, p16 ; Humans ; Jurkat Cells ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics ; Transcriptional Activation ; Transfection

OBJECTIVETo investigate the therapeutic effects of an early application of Chaiqin Chengqi Decoction (CQCQD) on severe acute pancreatitis (SAP) complicated with acute respiratory distress syndrome (ARDS).

METHODSForty patients of SAP-ARDS were equally randomized into the early-treated group (ET) and the late-treated group (LT), CQCQD was administered to them immediately and 3 days later after hospitalization respectively. Baseline materials in the two groups at the entry were insignificantly different (P > 0.05), and the same conventional Western medical therapy were available to them all. The Acute Physiology and Chronic Heath Evaluation II (APACHE I) scores, the incidence and sustained time of complications, the occurrence of infection, requirement of operation shifting on day 7, as well as the duration resided in hospital and mortality in patients were observed and compared.

RESULTSComparisons of the above-mentioned clinical indexes between groups showed that the APACHE II score was lower (5.1 +/- 2.0 scores vs 9.3 +/- 4.3 scores, P < 0.01); the incidence of shock was lesser (1/20 vs 7/19); the duration of ARDS, renal failure, cardiac insufficiency, hepatic dysfunction, cerebropathy and enteroplegia, as well as the duration in hospital and the requirement of operation shifting were all shorter significantly (P < 0.05) in the ET group than those in the LT group, but no statistical difference (P > 0.05) was shown in terms of the infection incidence and the mortality.

CONCLUSIONAn early application of CQCQD in the treatment of SAP could shorten the duration of complications and the couse of disease, lower the requirement of operation shifting. But further study with large samples for explore its impact on the infection incidence and the mortality is needed.

Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis, Acute Necrotizing ; complications ; drug therapy ; Phytotherapy ; Respiratory Distress Syndrome, Adult ; drug therapy ; etiology ; Time Factors ; Young Adult

Objective:To observe the lipid-lowering effect of different transdermal absorption enhancers applied to the herbal cake-partitioned moxibustion in hyperlipidemia model rabbits, and to explore the possible mechanism. Methods:Forty New-Zealand rabbits were randomly divided into 5 groups using the random number table method, with 8 rats in each group. Rabbits in the blank group were fed routinely with normal diet; rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model. Rabbits in the blank and the model groups were not treated. After the model was prepared, rabbits in the non-transdermal absorption enhancer group received herbal cake-partitioned moxibustion without transdermal absorption enhancer; rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively. After 4 weeks of treatment, serum was collected for enzyme-linked immunosorbent assay (ELISA), and the liver tissues were isolated for immunohistochemistry, quantitative polymerase chain reaction (qPCR) and Western-blotting (WB) detection. Results: Serum ELISA results showed that leptin was significantly decreased in the model group compared with the blank group (P＜0.05); compared with the model group, leptin was significantly increased in the non-transdermal absorption enhancer, the laurocapram and the borneol groups (all P＜0.05); compared with the non-transdermal absorption enhancer group, leptin was significantly increased in the laurocapram group and the borneol group (both P＜0.05); there was no significant difference in leptin between the laurocapram and the borneol groups (P＞0.05). The qPCR results of rabbit liver tissues showed that the mRNA expressions of leptin, Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in the model group were significantly lower than those in the blank group (all P＜0.05); compared with the model group, the mRNA expressions of leptin, leptin receptor (LR), JAK2 and STAT3 in the non-transdermal absorption enhancer, the laurocapram and the borneol groups were significantly increased (all P＜0.05); compared with the non-transdermal absorption enhancer group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the laurocapram and the borneol groups were significantly increased (all P＜0.05); compared with the laurocapram group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the borneol group were significantly increased (P＜0.05). The trend of immunohistochemistry and WB detection results was basically consistent with the qPCR assay results. The immunohistochemistry and WB detection results of phosphorylated JAK2 (phospho-JAK2) and phosphorylated STAT3 (phospho-STAT3) were basically consistent with those of JAK2 and STAT3. Conclusion: The molecular expression of Leptin/JAK2/STAT3 pathway in the hyperlipidemia model rabbits was decreased. The molecular expression of Leptin/JAK2/STAT3 pathway was significantly increased after the herbal cake-partitioned moxibustion. The application of laurocapram and borneol, as transdermal absorption enhancers, in the herbal cake-partitioned moxibustion could more obviously up-regulate the factors of the Leptin/JAK2/STAT3 lipid-regulating pathway than the herbal cake-partitioned moxibustion alone.

AIM:To study the dynamic alteration of low-density lipoprotein receptor ( LDLr) expression after exposure to hepatocyte growth factor (HGF) in human Tenon's capsule fibroblasts (HTFs).METHODS: HTFs were stimulated with HGF at different concentrations (0, 10, 20, 40, 80 and 160μg/L) for 12, 24, and 48 h.The viability of HTFs was analyzed by MTT assay .The expression of LDLr at mRNA and protein levels were analyzed by real-time PCR and Western blot .RESULTS:The expression of LDLr at mRNA and protein levels was positively correlated with the viability of HTFs.HGF promoted the viability of HTFs in a time-and concentration-dependent manner .At the same time , HGF pro-moted the expression of LDLr in the same manner .CONCLUSION:Exposure of HTFs to HGF induces LDLr expression at high level , suggesting that over-expression of LDLr on the HTFs may be a target receptor for controlled drug delivery , par-ticularly in anti-scarring therapy after glaucoma filtration surgery .

OBJECTIVESTo investigate targeted blockage of BCR/ABL oncoprotein mediated cell transformation by STAT5 decoy oligodeoxynucleotide (ODN), its effect on the growth and proliferation inhibition of K562 cells and the related molecular mechanisms.

METHODSSTAT5 decoy ODN, designed and synthesized in vitro, was transfected into K562 cells by cationic lipid. The cell growth curve and colony formation assay were used to reflect the growth and proliferation capacity of K562 cells, RT-PCR to detect the expression of three genes downstream STAT5.

RESULTSConfocal microscopy demonstrated that STAT5 decoy ODN was successfully transfected into K562 cells (95.2% positive cells). STAT5 decoy ODN inhibited the growth of K562 cells (inhibition rate 77.7%) and their colony formation capacity (Decoy ODN treated group 8.3% vs control group 35.7%, P < 0.05) after the treatment with STAT5 decoy ODN, the expressions of c-myc, bcl-X(L), cyclin D1 mRNA were down-regulated by 15.4%, 30.8%, 29.1%, respectively in the K562 cells.

CONCLUSIONSSTAT5 decoy ODN inhibits the growth and proliferation of K562 cells. The mechanisms may be that decoy ODN blocks the transcriptional activation potent of STAT5 and down-regulates the expression of these tumor related genes downstream STAT5.

Cell Proliferation ; Cyclin D1 ; genetics ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Humans ; K562 Cells ; Liposomes ; Microscopy, Confocal ; Oligodeoxyribonucleotides, Antisense ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; STAT5 Transcription Factor ; genetics ; physiology ; Transfection ; bcl-X Protein ; genetics

OBJECTIVETo explore the effects of vascular endothelial growth factor (VEGF) antisense phosphorothioated oligodeoxynucleotide (AS-ODN) on the expression of VEGF in human leukemic cell lines (HL-60 and K562 cells).

METHODSThe levels of VEGF mRNA and protein in leukemic cells incubated with VEGF AS-ODN were measured by RT-PCR, immunohistochemistry assay and ELISA. MTT test was used to examine the influence of the culture supernatant (CS) of VEGF AS-ODN treated leukemic cells on the proliferation of human umbilical vein endothelial cells (ECV304).

RESULTSAfter leukemic cells were treated with different concentrations (2.5 approximately 15.0 micro mol/L) of VEGF AS-ODN for 24 h, VEGF mRNA level in the cells decreased remarkably in a concentration dependent manner, no change was found in the VEGF missense ODN treated cells (MS-ODN). When the leukemic cells were treated with 5 micro mol/L VEGF AS-ODN for 24 h, VEGF protein level decreased greatly both in the cells and in the CS; and the proliferation stimulating effect of the treated CS on the ECV304 cells reduced. Meanwhile, there was no obvious change in VEGF protein and its effect in the VEGF MS-ODN treated group.

CONCLUSIONVEGF AS-ODN could inhibit VEGF expression in human leukemic cell lines in vitro.

Enzyme-Linked Immunosorbent Assay ; HL-60 Cells ; Humans ; Immunohistochemistry ; K562 Cells ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; analysis ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; genetics

OBJECTIVETo explore the diagnostic value of pulse transit time (PTT) in children with sleep disordered breathing(SDB).

METHODSForty eight randomly selected snorers (2 - 13 years) with SDB were examined by PSG and PTT in the same time. Data obtained were analyzed by different technicians respectively. Statistics and analysis of the data were performed.

RESULTSApnea hypopnea index (AHI), obstructive apnea index (OAI), the lowest oxygen and micro-arousal index were obtained by PSG and PTT. The results was described as M [25 percentile; 75 percentile]: 4.9[1.3;10.1], 4.6[1.5;11.8]; 1.2[0.7;4.9], 1.3[0.6;5.0]; 0.93[0.85;0.95], 0.93[0.84;0.95]; 14.5[12.6;16.4], 26.0[17.4;30.6]. The difference of AHI, OAI, and the lowest oxygen were not significant (P > 0.05), while the PTT arousal index detection rate was higher than PSG (Z = -5.19, P < 0.01). There was no significant difference in the diagnosis of obstructive sleep apnea-hypopnea syndrome (OSAHS) and determination of degree of patient's condition (P > 0.05). PTT could identify upper airway resistance syndrome in children without OASHS.

CONCLUSIONSBoth methods can be used to diagnose SDB. However, PTT is easy to use and suitable for the diagnosis of SDB in children, especially for UARS.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Polysomnography ; Pulse ; Sleep Apnea Syndromes ; diagnosis ; physiopathology

OBJECTIVETo analyze the similarities and differences of the clinical manifestations between the children with upper airway resistance syndrome (UARS) and obstructive sleep apnea-hypopnea syndrome (OSAHS), and to explore the clinical features and characteristics of sleep respiratory parameters.

METHODSUsing the double-blind method, all children were diagnosed as UARS or OSAHS through the polysomnography test and the results of all children were analyzed by a sleep technician and an otolaryngologist. Another ENT doctor recorded their clinical and physical examination in detail.

RESULTSPolysomnography showed that the apnea-hypopnea index (AHI) and the lowest oxygen in 253 children with OSAHS were 3.60[2.00;7.55] times/h and 0.90[0.85;0.91], and were 0.90[0.50;1.10] times/h and 0.95[0.92;0.96] in 102 children with UARS, the difference of the two groups by rank test was statistically significant. The proportion of UARS and OSAHS was more common during preschool period than during school-age period. The chief complaint in two groups was sleep snoring, and the main symptoms were sleep restless, attention deficit/hyperactivity and breath with mouth open. The incidence rate of above symptoms were as follows, 94.1%, 72.5%, 62.7% and 37.3% in children with UARS, 92.9%, 78.7%, 57.7% and 45.5% in children with OSAHS. The difference was not significant by chi-square test (P>0.05). Tonsil and adenoid hypertrophy were also observed in the two groups, the difference was not significant (chi2 = 0.27, P= 0.87). However, the children with OSAHS were more apt to have the sleep apnea than with UARS, the difference was statistically significant (chi2 = 34.07, P<0.001).

CONCLUSIONSThe clinical manifestations of two groups are similar, the difference between UARS and OSAHS can not be determined by the patient's clinical performance. Sleep apnea can be more easily observed in children with OSAHS than that in UARS, the final diagnosis is based on polysomnography.

Adolescent ; Airway Resistance ; Child ; Child, Preschool ; Double-Blind Method ; Female ; Humans ; Male ; Polysomnography ; Sleep Apnea, Obstructive ; epidemiology ; physiopathology

OBJECTIVETo explore the correlation between periodic limb movement index (PLMI) and the apnea-hypopnea index (AHI), apnea index (AI), hypopnea index (HI) and lowest oxygen saturation (LSaO₂) in sixty-four children with sleep-disordered breathing (SDB).

METHODSBetween March 2008 and May 2009, sixty-four children suspected of OSAHS underwent overnight polysomnogram monitoring in our medicine sleep center. OSAHS was diagnosed according to the general criterion. Sixty-four children were divided into two groups. Thirty children were diagnosed as OSAHS and 34 children were diagnosed as primary snoring (PS, 32 children) or upper airway resistance syndrome (UARS, 2 children). The difference of PLMI and periodic limb movement index during sleep associated with arousals (PLMI-arousal) were compared between the two groups. Besides this, the correlation between PLMI, periodic limb movement index during sleep associated with arousals and AHI, AI, HI and LSaO₂ were also analyzed in all SDB children. Furthermore, all SDB children were divided into two groups according to PLMI (< 5 events/h vs ≥ 5 events/h). AHI, AI, HI, LSaO₂ and sleep structure were compared between the two groups.

RESULTS(1) The difference of PLMI and PLMI-arousal between the children with OSAHS and children with other SDB types (PS and UARS) were not significant (z value, -1.279, -1.490; P value, 0.201, 0.136, respectively). (2) The increased sleep stage I was significant as being compared between the two groups (< 5 events/h vs ≥ 5 events/h, t = -2.16, P < 0.05). However, other sleep stages and sleep efficiency were not significantly different (P value, all > 0.05). (3) The difference of HI, AI, AHI, arousals index (ArI) and LSaO₂ were not significant between the two groups (< 5 events/h vs ≥ 5 events/h, P value, all > 0.05). (4) PLMI and PLMI-arousal were not correlated with AHI, HI, AI, AHI and LSaO₂ (Spearman rank correlation analysis).

CONCLUSIONSPLMS may be independent of SDB and PLMS had a little influence on sleep structure.

Adolescent ; Child ; Child, Preschool ; Extremities ; physiopathology ; Female ; Humans ; Male ; Movement ; Polysomnography ; Sleep ; Sleep Apnea Syndromes ; physiopathology

OBJECTIVETo analyze the effect of three-dimensional (3D) laparoscopic total thyroidectomy combined with central lymph node dissection for thyroid cancer and its effect on the inflammatory response of the patients.

METHODSThe clinical data were analyzed in 90 patients with thyroid cancer undergoing radical thyroidectomy at our hospital between September, 2013 to April, 2016, including 30 receiving 3D laparoscopic surgeries, 30 with 2D laparoscopic surgeries and 30 with open surgeries. The surgical data, postoperative adverse reactions and the impact of the surgeries on the inflammatory responses of the patients were compared among the 3 groups.

RESULTSCompared with the open surgery and 2D laparoscopic surgery, 3D laparoscopic surgery was associated with lowered blood loss during the surgery and a lowered incidence of adverse reactions. The operation time in 3D group was significantly shorter than that in 2D group (P<0.05), but the total hospitalization expenses were similar between the two groups. The postoperative drainage volume did not differ significantly between the 3D group and the other two groups. The postoperative hospital stay, number of lymph nodes dissected, positivity rate of lymph nodes and the inflammatory response showed no significant differences among the 3 groups (P>0.05).

CONCLUSION3D laparoscopic total thyroidectomy combined with central lymph node dissection is safe and effective and reduces intraoperative blood loss and perioperative adverse reactions without significant influence on inflammatory response in patients with thyroid cancer.