1.Clinical significance of STMN1 expression in cervical cancer and effect of inhibition of its expression on viability and apoptosis of cervical cancer Si-Ha cells
Zhi-Hong CHAI ; Ling-Xiao YING ; Yi-Hong PAN ; Xian-Li CAI ; Wei-Min KONG
Chinese Journal of Pathophysiology 2018;34(6):1119-1123,1128
AIM:To investigate the clinical significance of stathmin 1 (STMN1) expression in cervical cancer and the influence of its expression on the viability and apoptosis of cervical cancer cells. METHODS:Western blot was used to detect the protein expression of STMN1 in cervical cancer tissues, and the relationship between the expression and clinical characteristics of cervical cancer was analyzed. STMN1-siRNA was transfected into cervical squamous-cell carcino-ma SiHa cells. The protein levels of STMN1, STAT3, p-STAT3 and survivin were determined by Western blot after trans-fection for 48 h. The cell viability was measured by MTT assay. The apoptosis was analyzed by flow cytometry. DCFH-DA probe was used to detect the level of reactive oxygen species (ROS). RESULTS:The protein expression of STMN1 in cer-vical cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The STMN1 protein expression level was not correlated with age and histological types of cervical cancer patients, but was related to clinical stage, histo-logical differentiation and lymph node metastasis ( P<0.01). Transfection with STMN1-siRNA significantly reduced the expression of STMN1 in SiHa cells. Compared with control group, the cell viability in STMN1-siRNA group was significant-ly decreased, the apoptotic rate and ROS content were increased, and the protein levels of p-STAT3 and survivin were down-regulated (P<0.01). However, no significant difference of the STAT3 protein level was observed between STMN1- siRNA group and control group. CONCLUSION:STMN1 is highly expressed in cervical cancer, and its expression is re-lated to clinical stage, histological differentiation and lymph node metastasis. Inhibition of STMN1 expression reduces the viability and promotes apoptosis of cancer cells by down-regulating STAT3 signaling pathway.
2.The inhibitory effect of chronic lead exposure on CA1 LTP and ERK2 activity in rat hippocampal area.
Jing YANG ; Li-guang SUN ; Zhi-hong ZHONG ; Kui CHAI ; Hong-xin WANG
Chinese Journal of Applied Physiology 2006;22(3):326-328
AIMTo investigate the effect of chronic lead exposure on rat hippocampal CA1 LTP and alpha-Ca2+ /calmodulin-dependent protein kinase II (alpha-CaM K II) activity in vivo.
METHODSA stimulus bipolar electrode was placed on the Schaffer/Commissural fibers, with extra cellular microelectrode technique to record the population spike (PS) in the CA1 pyramidal, and we observed the changes of PS amplitude before and after the high frequency stimulation (HFS) of lower, mid and higher level lead exposure groups and the control group, respectively. The hippocampal CA1 alpha-CaM K II activity was determined by Western blots by using phosphorylation antibody.
RESULTSThe average changes of PS after HFS of the control group, the lower, mid and higher level lead exposure groups were 162.5%, 105.2%, 86.8%, 83.0%, respectively (P < 0.01 vs. control). Defined control a-CaM K II activity as 100, that the lower, mid and higher level lead exposure groups were 62.0 +/- 3.7, 50.8 +/- 4.0, 43.3 +/- 4.1 (P < 0.01).
CONCLUSIONChronic lead exposure can inhibit CA1 LTP in vivo, and the decrease of alpha-CaMK II activity may play an important role in this mechanism.
Animals ; CA1 Region, Hippocampal ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Lead ; toxicity ; Long-Term Potentiation ; Rats ; Rats, Wistar ; Toxicity Tests, Chronic
3.Influence of carbachol on intestinal dysfunction after traumatic or burn injury.
Wei-hong CAO ; Jia-ke CHAI ; Sen HU ; Hong-ming YANG ; Tian-jun SUN ; Xiao-fang ZOU ; Zhi-yong SHENG
Chinese Journal of Burns 2006;22(3):168-171
OBJECTIVETo investigate the influence of enteral administration of carbachol on the intestinal dysfunction of both severely burn patients and rabbits with partial intestinal ischemia/reperfusion (I/R) injury.
METHODSSeventy-five white rabbits were inflicted with I/R injury and randomized into intestinal I/R (I, n=25), carbachol [C, n=25, with 3g/L carbachol (3 mg/kg) injection into duodenum 1 h after SMA occlusion] and sham operation (SO, n=25, with SMA isolation but no occlusion) groups, and 5 other as normal controls. The blood flow of intestinal mucosa was detected before and after SMA occlusion or admission of carbachol. Changes in diamine oxidase (DAO), D-lactate, xylopyranose absorption, blue dextran discharging time were measured at 2, 4, 6, 8, 24, 48, 72 h after SMA occlusion. In addition, eight severe burn patients with TBSA of 84 +/- 12% were enrolled in the study, and carbachol (15 microg/kg) was administered to patients when abdominal distension or bowel sound was lower than 2 times/min, then the number of abdominal distension and bowel sounds per minute were observed.
RESULTSThe blood flow in intestinal mucosa of rabbits without SMA occlusion was (102 +/- 5) PU, reduced to (48 +/- 6) PU after SMA occlusion, and increased to (77 +/- 3) PU after injection of carbachol. The plasma DAO activity and D-lactic acid content in I group began to increase 4 hours after SMA occlusion, and they reached the peak 24 hours after SMA occlusion (4.63 +/- 0.27 U/ml, 7.9 +/- 2.4 mg/L) , after that they decreased gradually, but still higher than the normal value (0.89 +/- 0.14 U/ml, 2.0 +/- 1.1 mg/L, P < 0.05). In carbachol group, data showed the same trends as that in intestine I/R group with lower values, while no obvious changes were in sham operation group (P > 0.05). The content of D-lactic decreased dramatically 2 hours after D-lactic administration in both I and C groups, increased 6 hours after SMA occlusion, then decreased gradually, but it in C group was always higher than normal values, and little fluctuation was in sham operation group. There was no blue dextran discharge 2 hours after SMA occlusion. The discharging distance increased 6 hours later, but it was obviously shorter than the normal value 24 hrs after operation (P < 0.05) , then it returned to normal 48 to 72 hrs after operation. In the C group, blue dextran discharge was found immediately after its injection, with obvious increase in the discharging distance to peak value (43 +/- 6 cm) 6 hours after injury, and returning to normal (28 +/- 3 cm) gradually. In severe burned patients, the bowel sounds was (1.6 +/- 1.1) per minutes before carbachol administration, then increased dramatically to (6.9 +/- 1.7) per minutes 10 mins after administration, reached to a higher level 30 minutes after administration (8.3 +/- 2.4 ) times/min, and it maintained to (6.1 +/- 1.3) times/min 1 hour after administration. Abdominal distension was ameliorated 2 hours after carbachol administration, six patients were able to defecate.
CONCLUSIONEnteral administration of Carbachol can increase the blood flow of intestine mucosa, help to improve the movement, absorption and barrier functions of intestine, and ameliorate intestinal dysfunction in patients with severe burns.
Adolescent ; Adult ; Animals ; Burns ; drug therapy ; physiopathology ; Carbachol ; therapeutic use ; Disease Models, Animal ; Female ; Humans ; Intestinal Mucosa ; blood supply ; metabolism ; Intestines ; physiopathology ; Male ; Rabbits ; Reperfusion Injury ; drug therapy ; physiopathology
4.The effects of early escharectomy on resting energy expenditure in the severely burned patients.
Jian-chuan GAO ; Jia-ke CHAI ; Hong-ming YANG ; Zhi-yong SHENG ; Dai-feng HAO ; Zhen-rong GUO
Chinese Journal of Burns 2004;20(1):37-39
OBJECTIVETo investigate the effect of early escharectomy on resting energy expenditure (REE) in severely burned patients dynamically with the metabolic monitoring and diagnostic system.
METHODSFifty-six adult male patients with severe burns were divided into early escharectomy (group A, n = 39, escharectomy within 5 PBDs) and non-early escharectomy (group B, n = 17, escharectomy after 5 PBDs) groups. The wounds of full thickness and deep partial thickness burn in the two groups were all excised and covered with allogeneic skin and autologous micro-skin in the first operation. The changes in REE were observed dynamically at the bedside of the patients with the metabolic monitoring and diagnostic system. The plasma contents of IL-6, IL-8, TNF-alpha and LPS from 9 patients in group A and 7 in group B were also determined dynamically.
RESULTSAll patients survived. The REE in both groups was elevated markedly, but REE in group A was lower compared with group B before and after escharectomy within 14 days. (P < 0.05). The plasma level of IL-6, IL-8, TNF-alpha and LPS in group A were obviously lower than those in group B (P < 0.05).
CONCLUSIONThe hypermetabolic response of burn patients with severe burns could be lowered by early escharectomy, and it seemed to be related to the decrease of the release of proinflammatory mediators.
Adult ; Basal Metabolism ; Burns ; metabolism ; physiopathology ; surgery ; Humans ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Lipopolysaccharides ; blood ; Male ; Postoperative Care ; Time Factors ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; metabolism
5.Effects of insulin on proteolysis of cultured rabbit skeletal muscular myotubes in vitro.
Chuan-an SHEN ; Jia-ke CHAI ; Zhi-yong SHENG ; Hong-ming YANG ; Hui-man YIN ; Rui FENG
Chinese Journal of Burns 2006;22(4):262-265
OBJECTIVETo study the effects of insulin on the proteolysis of cultured rabbit skeletal muscular myotubes in vitro, and their possible mechanisms.
METHODSMuscles of lower limbs of juvenile rabbits were isolated for tissue-block culture. After passage, myoblasts were formed and fused into myotubes. Then the protein in myotubes was radiolabelled with L-[ 3,5-3H] tyrosine. The myotubes were cultured in DMEM medium containing 100 nmol/L insulin (n = 24, group B) , 100 nmol/L dexamethasone (n = 24, group C) , 100 nmol/L insulin and 100 nmol/L dexamethasone (n = 24, group D) , no insulin or dexamethasone (n =24, group A), respectively. Twenty-four hours after culture, the L-[3,5-3H] tyrosine content in culture medium and cells were determined, and the degradation rates of protein were calculated. The mRNA expressions of ubiquitin and protease C2 subunit were determined by Northern blot.
RESULTSThe degradation rates of myotube protein in group A(0. 38+/-0.04) was obviously lower than that in group C (0.50+/-0.03, P <0.01), but it was obviously higher than that in group B(0. 35+/-0.03, P <0.05). Though the degradation rates of myotube protein in group D (0.41+/-0. 03) was evidently lower than that in group C ( P < 0.01) , it was still higher than that in group A( P < 0.05 ). The mRNA expressions of ubiquitin and protease C2 subunit in group A ( the scale: 2. 4 kb ubiquitin was 0. 82+/-0. 15, 1. 2 kb ubiquitin was 0. 60+/-0. 10, C2 subunit was 0. 75+/-0. 16) was obviously lower than that in group C ( the scale: 2.4 kb ubiquitin was 2. 15+/-0. 23, 1.2 kb ubiquitin was 1.50+/-0. 14,C2 subunit was 1.50+/-0. 13 , P <0. 01) , but it in group D was lower than that in group C (the scale: 2. 4 kb ubiquitin was 1. 25+/-0. 17, 1. 2 kb ubiquitin was 0. 85+/-0. 09, C2 subunit was 0. 90+/-0. 15, P <0. 01) , and it was similar to that in group B (the scale: 2.4 kb ubiquitin was 0. 85+/-0.07, 1.2 kb ubiquitin was 0. 65+/- 0. 12, C2 subunit was 0. 76 +/-0. 09, P > 0. 05).
CONCLUSIONThe effects of insulin on the activity of ubiquitin-proteasome pathway and the proteolytic rate in normal myotubes were relatively weak. However, insulin can significantly inhibit the effects of dexamethasone on the gene expressions of ubiquitin system and the proteolytic rate in myotubes, but the mechanism needs further research.
Animals ; Cells, Cultured ; In Vitro Techniques ; Insulin ; pharmacology ; Male ; Muscle Fibers, Skeletal ; drug effects ; metabolism ; Muscle Proteins ; metabolism ; Rabbits ; Ubiquitin ; metabolism
6.Preparation and antibacterial activity of quaternary ammonium salt monomers.
Yu-Hong XIAO ; Ji-Hua CHEN ; Ming FANG ; Xiao-Dong XING ; Fang LI ; Zhi-Guo CHAI
Chinese Journal of Stomatology 2008;43(6):370-373
OBJECTIVETo prepare three quaternary ammonium salt (QAS) monomers, and to compare their antibacterial activities against four oral bacterial strains.
METHODSThree antibacterial monomers [methacryloxyethyl benzyl dimethyl ammonium chloride (DMAE-BC), methacryloxyethyl m-chloro benzyl dimethyl ammonium chloride (DMAE-m-CBC), methacryloxyethyl cetyl dimethyl ammonium chloride (DMAE-CB)] were synthesized according to the general structure of target monomers. Their antibacterial effects were investigated using the broth dilution test on Gram-positive and Gram-negative bacterial strains (Streptococcus mutans, Streptococcus sanguis, Porphyromonas gingivalis, Prevotella melaninogenica ).
RESULTSThree different monomers were successfully obtained. All the tested bacterial strains were susceptible to the three monomers, among which DMAE-CB exhibited the lowest minimal inhibitory concentrations (MIC) ranging from 1.2 to 4.8 mg/L.
CONCLUSIONSAll these three QAS monomers have different antibacterial activities against four oral bacteria strains. The data indicate that DMAE-CB may be a candidate antibacterial agent for oral infectious diseases.
Anti-Bacterial Agents ; chemistry ; pharmacology ; Dental Materials ; chemistry ; pharmacology ; Microbial Sensitivity Tests ; Quaternary Ammonium Compounds ; chemistry ; pharmacology
7.Differentiation of human embryonic stem cells into neural stem cells in vitro
Feng-Guang LI ; Hui-Fang XU ; Hong CHEN ; Jing-Yan CHAI ; Bian-Zhi XING ; Yan-Qiang ZHAN ; Xiao-Qing LI ; Shou-Gang GUO ; Su-Ming ZHANG ;
Chinese Journal of Neurology 2005;0(07):-
Objective To explore a kind of simple and high efficient approach to differentiate human embryonic stem cells(hESCs)into neural stem cells(NSCs).Methods hESCs were cultured in bacterial culture dish filled with serum free medium to gain embryoid bodies.Then the mature embryoid bodies grew in the special medium including B27 and noggin by adherent culture to differentiate into NSCs. Results The hESCs kept floating in the bacterial culture dish and growing all the time and then formed mature embryoid bodies 7 to 10 days later.The embryoid bodies could be differentiated into highly pure (96.4%)nestin positive cells.And these cells were differentiated into all kinds of neural cells if cultured further.Conclusions This kind of method is less time-consuming,cheaper,and more efficient than those of the results in literatures reported.It affords very good source of seed cells for cell transplantation therapy in the future.
8.The expression of platelet-derived growth factor-A and its receptor in acute radiation-induced skin ulcer in rats.
Wei-hong CAO ; Jia-ke CHAI ; Zhi-xiang YANG ; De-wen WANG ; Zhi-yong SHENG
Chinese Journal of Burns 2005;21(5):359-362
OBJECTIVETo investigate the dynamic expression of platelet-derived growth factor-A (PDGF-A) and its receptor alpha (PDGFR-alpha) in different acute radiation-induced skin ulcers, and to explore the underlying mechanism involved in retarded healing of the ulcer.
METHODSThe model of acute radiation-induced skin ulcers in rats was replicated with 50 Gy 60Co gamma rays to the skin (radiation group, R, n = 55), rats with full - thickness skin excision wounds as control group (T, n = 55), and 5 normal rats to serve as normal control (NC) group. The expression of PDGF-A and PDGFR-alpha protein and PDGF-A mRNA was respectively assessed by means of histochemistry and in situ RT-PCR.
RESULTSNo PDGF-A expression was identified in the rat skin in NC group. The expression of PDGF-A and PDGFR were reduced in R group during inflammatory responsive and granulation formation periods (14 - 28 days after radiation, the IA value of PDGF-A varied from 14.0 +/- 1.2 to 20.3 +/- 1.2 compared with that in T group in which the IA value of PDGF-A at the same period (3 - 9 days after injury) varied from 20.0 +/- 1.6 to 28.3 +/- 1.0, and reduced gradually during scar formation period (55 days after radiation).
CONCLUSIONThe reduction of PDGF-A and PDGFR-expression may be partially involved in the mechanism of retarded healing of acute radiation-induced skin ulcers.
Animals ; Female ; Gamma Rays ; adverse effects ; Platelet-Derived Growth Factor ; biosynthesis ; Radiation Injuries, Experimental ; complications ; metabolism ; Rats ; Rats, Wistar ; Receptor, Platelet-Derived Growth Factor alpha ; biosynthesis ; Skin Ulcer ; etiology ; metabolism ; Wound Healing
9.The influence of CD14 genomic polymorphism on CD14 gene expression as well as protein release and its clinical significance in patients with extensive burns.
Jing LIN ; Yong-Ming YAO ; Zhi-Hong HUANG ; Yan YU ; Jing-min ZHU ; Jia-Ke CHAI ; Zhi-Yong SHENG
Chinese Journal of Surgery 2006;44(13):907-910
OBJECTIVETo investigate the influence of a lipopolysaccharide receptor CD14-159C/T genomic polymorphism on CD14 gene expression as well as protein release, and the relation of sepsis susceptibility and prognosis in patients with extensive burns.
METHODSThe study group consisted of 26 patients with burns covering more than 30% of the total body surface area. The CD14 gene polymorphism was determined by polymerase chain reaction (PCR) and subsequent HaeIII restriction enzyme digestion of the PCR products. Meanwhile, the association of CD14, TNF-alpha mRNA expression in leukocytes and soluble CD14 (sCD14) levels in serum with CD14-159 polymorphism as well as prognosis after burns was also studied.
RESULTSThe T allele frequencies in sepsis patients and non-survivors were higher than those in non-septic patients and survivors. The levels of CD14 mRNA, TNF-alpha mRNA expression and serum sCD14 were significantly different among patient groups with TT, TC, and CC genotypes. The above differences were also existed between survivors and non-survivors. CD14 mRNA expression was higher in heterozygotes (TT and TC) than in C homozygous patients (P < 0.05 or P < 0.01), and sCD14 level was higher in heterozygotes (TC) than C homozygous patients on day 7 postburn (P < 0.05). During the 28-day observation period, mean TNF-alpha mRNA expression was higher in patients homozygous for T allele than C homozygotes (P < 0.05). In addition, higher CD14 mRNA values were found in non-survivors compared to those in survivors on days 7 and 28 postburn (P < 0.05).
CONCLUSIONSCD14C-159T polymorphism might markedly influence CD14 mRNA expression and sCD14 levels, and it seems to be associated with sepsis susceptibility and prognosis in patients with extensive burns. The T allele could be a genetic risk marker of adverse prognosis.
Adult ; Burns ; complications ; metabolism ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lipopolysaccharide Receptors ; biosynthesis ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Prognosis ; RNA, Messenger ; genetics ; Sepsis ; etiology ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics
10.Influence of Shenfu Injection on the quality of life of lung cancer patients receiving chemotherapy.
Shun-qin LONG ; Gui-ya LIAO ; Wen-feng HE ; Bin WANG ; Hong DENG ; Hai-bo ZHANG ; Xiao-shu CHAI ; Jiao-zhi CAI ; Wan-yin WU
Journal of Southern Medical University 2011;31(12):2090-2092
OBJECTIVETo evaluate the influence of Shenfu Injection (SHF) on the quality of life of patients with advanced non-small cell lung cancer (NSCLC) receiving chemotherapy.
METHODSA total of 133 patients with NSCLC receiving at least two cycles of chemotherapy with taxol plus cisplatin (TP)/vinorelbine plus cisplatin (NP) or gemcitabine plus cisplatin (GP) were randomized into SHF pre-treatment group (with SHF given only in the first cycle) and SHF post-treatment group (with SHF given only in the second cycle). The Quality of Life Questionnaire-Core 30 (QLQ-C30) and the Functional Living Index-Cancer (FLIC) were used to evaluate the quality of life of the patients after the treatments.
RESULTSBoth of the groups showed improved quality of life after the treatments (P<0.01), but the improvements were more obvious in SHF pre-treatment group (P<0.05). SHF showed favorable effects in relieving such adverse effects as fatigue, nausea, vomiting and diarrhea associated with the chemotherapy.
CONCLUSIONSHF can improve the quality of life in NSCLC patients receiving chemotherapies.
Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Cisplatin ; administration & dosage ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Nausea ; prevention & control ; Paclitaxel ; administration & dosage ; Phytotherapy ; Quality of Life ; Surveys and Questionnaires ; Vinblastine ; administration & dosage ; analogs & derivatives ; Vomiting ; prevention & control