[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.

Objective To explore the basic ingredients of the tree shrew’ s( Tupaia belangeri) milk and compare with the dairy ingredients of other milks.Methods We select ten seed tree shrews after delivery ( 1 ~21 ) d with lactation mother tree shrews, and use artificial passive breastfeeding method let the young tree shrews suck breast milk,we took the milk from the young tree shrews in the stomach, directly using aseptic operation with a syringe immediately, once every two days, for consecutive three to five times, and a total of 18 mL milk was taken from each seed tree shrew.Then the milk was detected according to the national standard method for component testing.Results The total solid content of the tree shrew’ s milk was 43.63%, including 26.01%of fat, 10.41%of protein, 0.45% of lactose and 0.99%of ash content.Compared with cow's milk, the tree shrew’ s milk contained 3.36 times of total solid contents, 1.24 times of ash, 2.74 times of protein, 6.67 times of fat, and 0.09 times of lactose.Compare with baby formula milk, the tree shrew’ s milk contained 1.44 times of total solid contents, 0.20 times of ash, 0.58 times of protein, 1.53 times of fat, and 0.06 times of lactose.The trace mineral composition of the tree shrew’ s milk showed that the calcium, phosphorus, potassium, sodium, magnesium, and iron contents were 1.83 times, 2.73 times, 1.25 times, 1.93 times, 1.28 times, and 1.48 times higher than those in the cow's milk, and were 0.66 times, 0.85 times, 0.34 times, 0.26 times, 0.85 times, 0.24 times lower than those in baby formula milk.Conclusions The main nutrients of tree shrew’ s milk is of high fat, high protein and low sugar, and it can provide a basis for tree shrews artificial brood and breeding work.

Objective: It is to investigate the efficacy of the blood-activating and orifice-opening method with acupuncture and moxibusion for treatment of vertebroarterial cervical spondylopathy. Methods: 176 cases were divided into the treatment group (89 cases) and control group (87 cases) randomly. Acupuncture on points Geshu (BL 17), Fengchi (GB 20) and Bailao (Ex-HN) was applied, and meanwhile moxibustion on points Baihui (GV 20) and Shangxing (GV 23) was applied in the treatment group, while Flunarizine was orally taken in the control group. Results: The blood-activating and orifice-opening method with acupuncture and moxibustion could significantly improve the comprehensive curative effect clinically, increase the subjective index score and the sign score significantly, and change hemodynamics obviously. Its effect was better than that of the control group. It was also found that its follow-up effect was significantly better that of the control group. Conclusion: The blood-activating and orifice-opening method with acupuncture and moxibustion has a definite effect on vertebroarterial cervical spondylopathy.

OBJECTIVETo evaluate the effects of periodontal treatment on the clinical response, systemic inflammatory parameters, and metabolic control of type 2 diabetes patients with moderate to severe periodontitis.

METHODSA total of 56 patients with mean clinical attachment level (CAL)>3 mm were included in the subgroup analysis. A repeated-measures ANOVA (group factor: treatment group and control group; time factor: initial visit, 1.5, 3, and 6 months) was used to analyze the probing depth (PD), CAL, bleeding on probing (BOP), high-sensitivity C-reactive protein (hsCRP), glycated hemoglobin (HbA1c), and fasting plasma glucose.

RESULTSSignificantly lower PD (F=62.898, P-0.000), CAL (F=51.263, P-0.000), BOP (F=75.164, P=0.000), hsCRP (F=6.391, P=0.010), HbA1c(F=4.536, P=0.011), and fasting plasma glucose level (F= 3.073, P=0.031) were observed after therapeutic periodontal improvement. The inter-group differences for PD (t=-2.050, P=0.045), BOP (t=-4.538, P=0.000), and hsCRP (t=-2.261, P=0.028) were statistically significant after therapy.

CONCLUSIONNon-surgical periodontal treatment can effectively improve periodontal status, circulating inflammatory status, and metabolic control of diabetic patients with moderate to severe periodontitis.

C-Reactive Protein ; Chronic Periodontitis ; Diabetes Mellitus, Type 2 ; Glycated Hemoglobin A ; Humans ; Periodontitis

N-type voltage-gated calcium (Ca²⁺) channels (N-type VGCC, Ca_V2.2) mediate Ca²⁺ influx in response to action potential at the presynaptic terminal, and play an important role in synaptogenesis, neurotransmitter release and nociceptive signal transduction. It is a new target for the development of drugs for the treatment of neuralgia (chronic pain) and other major diseases. Due to the difficulty of calcium channel expression in vitro and the detection of channel current, there is a great lack of new drug screening models. In this study, we established and optimized the electrophysiological drug screening model using Xenopus laevis oocytes for the recombinant expression of Ca_V2.2 in vitro (this study were reviewed and approved by the Ethics Committee of Guangxi University, approval number: GXU-2023-0249). Firstly, the linear plasmids encoding cDNA of major subunit α1B and auxiliary subunits α2δ1 and β3 of rat Ca_V2.2 were used as templates for in vitro transcription to generate their related mRNA (cRNA), after which three kinds of cRNA were injected into Xenopus laevis oocytes at the mass ratio of 2∶1∶1 for expression. The two-electrode voltage clamp (TEVC) technique was used to detect the inward current produced by Ca_V2.2. At the same time, the expression conditions of Ca_V2.2 were optimized, and its gating function was characterized from the aspects of channel activation and inactivation. The results showed that 3-5 days after cRNA microinjection, stable Ca_V2.2-mediated barium ion (Ba²⁺) currents were successfully detected. The interference of endogenous potassium channels and Ca²⁺-activated chloride channels can be eliminated by tetraethylammonium hydroxide (TEAOH) and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (BAPTA-AM) treatment. The maximum potential for Ca_V2.2 activation is 0 mV, and the current reverses to be outward when the membrane potential is greater than +50 mV. By fitting the steady-state activation and inactivation curves, the half-maximal activation potential and half-maximal inactivation potential of Ca_V2.2 are identified as -15.9 and -60.2 mV. In this study, a stable Ca_V2.2 expression system was established based on Xenopus laevis oocytes. The in vitro expression system can provide a new way for the screening of Ca_V2.2 active compounds or lead drugs.

Objective To investigate the mRNA expression and methylation status of IL-13 receptor(IL-13R)α1 gene in peripheral T lymphocytes of patients with systemic lupus erythematosus(SLE).Methods Venous blood samples were obtained from 10 SLE patients(5 in active phase,5 in inactive phase)and 6 normal human controls.CD4+ and CD8+ T cells were isolated from these samples via magnetic activated cell sorting(MACS).Real-time quantitative PCR was used to test the mRNA expression of IL-13Rα1 gene,and methylation specific PCR to detect the methylation status.Results The expression level of IL-13Rα1 mRNA was 2.224±0.251,1.712±0.132.and 1.104±0.044 in CD4+ T cells of active SLE patients,inactive SLE patients and controls,respectively;the difference between the three groups was statistically significant(all P＜0.05).The expression level of IL-13Rα1 mRNA in CD8+T cells was significantly higher in active SLE patients than that in the normal controls(1.672±0.142 vs 1.238±0.106,P＜0.05),while no difference was noted between inactive and active SLE patients or normal controls.The methylation index of IL-13Rα1 gene was 0.454±0.023.0.635±0.065.0.844±0.097 in CD4+T cells of active SLE patients,inactive SLE patients and normal controls,respectively,and the difference between the three groups was significant(all P＜0.05),while no significant difference was observed in the methylation index in CD8+T cells among these groups(P＞0.05).The IL-13Rα1 mRNA expression in CD4+T and CD8+T cells was positively correlated with SLE disease activity index(SLEDAI)score(r=0.79,0.76,P=0.007,0.02 respectively).A negative correlation was found between the methylation level Of IL-13Rα1 in CD4+T cells and SLEDAI score(r=-0.89.P＜0.0 1).as well as between the IL-13Rα1 mRNA expression and its methylation level(r=-0.84,P＜0.0 1).Conclusion The development of SLE may be related to the overexpression of IL-13Rα1 gene induced by DNA hypomethylation in T cells.

Objective To explore the mechanism of opioid-psychic dependence involving the aspects of pre-receptor and receptor by observing the changeable expressions of glutamate neurotransmitter and NR2B of N-methyl-D-aspartic acid (NMDA) receptor in the neuroanatomical circuit of ventral temental area, nucleus accumbens and prefrontal cortex (VTA-Nac-PFC) of rats subjected to morphine-induced conditioned place preference (CPP). Methods The models of CPP were validated by escalating doses of morphine in rats (n=16). The colorimetry and immunohistochemistry ways were applied to detect the contents of glutamic acid and the expression level of NR2B in VTA, Nac and PFC. Results As compared with those in the control group physiological saline), the prolonged detention time of white compartment in the model group was notably observed (P<0.05), and increased content of glutamic acid and expression level of NR2B in fields of VTA, Nac and PFC in the model group were significantly detected (P<0.05). Conclusion Increased level of giutamic acid and expression level of NR2B in nuroanatomieal circuit of VTA, Nac and PFC could play key roles in inducing morphine-psychic dependent rats.

BACKGROUND AND OBJECTIVEAs chemotherapy is generally used in the clinical treatment of cancer, the problem of multidrug resistance (MDR) of tumors against the chemotherapeutic agents becomes more and more serious. It has been the major cause for the failure of the chemotherapy. We detected the expressions of beta-catenin and tumor drug resistance related proteins, MRP2, P-gp, and Bcl-2, in esophageal squamous cell carcinoma (ESCC) to explore their function and correlation in the occurrence and development of MDR in ESCC.

METHODSWe used the tissue microarray technique, immunohistochemistry, and image analysis methods to detect the expressions of MRP2, P-gp, beta-catenin, and Bcl-2 proteins and analyze their relationships with clinical data in a ESCC tissue microarray consisting of 582 specimens of ESCC, 294 specimens of normal mucosa, 92 specimens of basal cell hyperplasia, and 87 specimens of dysplasia adjacent to cancer tissue.

RESULTSThe integral optical density (IOD) of MRP2 and Bcl-2, which was 195.7 +/- 175.9 x 10(3)) and 90.5 +/- 112.5 x 10(3)), respectively, was significantly higher in ESCC than in normal mucosa, which was 104.8 +/- 86.1 x103) and 25.2 +/- 46.6 x 10(3)), respectively (P < 0.01). The IOD of P-gp and beta-catenin, which was 57.7 +/- 75.5 x 10(3)) and 32.0 +/- 47.0 x 10(3)) respectively, was significantly lower in ESCC than in normal mucosa, which was 114.8 +/- 106.6 x 10(3)) and 46.1 +/- 35.7 x 10(3)), respectively (P < 0.01). According to the following order, well differentiated moderately differentiated poorly differentiated, the IOD of MRP2 increased in ESCC (P < 0.01). The IOD of beta-catenin was higher in poorly differentiated ESCC than in well or moderately differentiated ESCC (P < 0.01). The IOD of Bcl-2 was lower in well differentiated ESCC than in poorly and moderately differentiated ESCC (P < 0.01). The IOD of beta-catenin and Bcl-2 was higher in the ESCC of specimens with infiltration depths that were in membrane mucosa than those in the muscular layer or serous coat (P < 0.01). The IOD of Bcl-2 was significantly higher in cases with lymph node metastasis than in cases without (P < 0.01). Positive correlations which were respectively between the expressions of P-gp and MRP2, the expressions of P-gp and Bcl-2 were found (r = 0.288 and r = 0.253, respectively, P < 0.01).

CONCLUSIONSMRP2, P-gp, and Bcl-2 may be taken as prognostic factors for MDR of ESCC. beta-catenin may play an important role in carcinogenesis and progression of ESCC.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Multidrug Resistance-Associated Proteins ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Young Adult ; beta Catenin ; metabolism

OBJECTIVETo study the acting mechanism of anti-morphine conditioned place preference (CPP) between aqueous extract of Corydalis yanhusuo and L-THP and compare their effects.

METHODThe CPP model was established by injecting morphine in rats with a increasing dose for 10 days, with the initial dose of 10 g x kg(-1) and the final dose of 100 g x kg(-1), 10 mg x kg(-1) was increased each day, thus 100 mg x kg(-1) was injected by d 10. Having been treated with differential doses (2, 1 and 0.5 g x kg(-1)) of C. yanhusuo (containing L-THP: 0.153, 0.077 and 0.038 mg x kg(-1) respectively) and L-THP (3.76, 1.88 and 0.94 mg x kg(-1)) for six days, the CPP effect in rats was detected. Both colorimetry and immunohistochemistry methods were adopted to detect the content of glutamate neurotransmitter in each brain region and the expression of NR2B in VTA-NAc-PFC neuroanatomical circuit.

RESULTCompared with the physiological saline treatment group, C. yanhusuo (2, 1 g x kg(-1)) and L-THP (3.76 and 1.88 mg x kg(-1)) groups showed a notably shorter retention period of rats in white boxes (morphine-accompanied boxes) (P < 0.05 or P < 0.01) and remarkably lower glutamic acid content in VTA, NAc and PFC and NR2B expression.

CONCLUSIONBoth C. yanhusuo and L-THP can substantially inhibit the effect of morphine CPP, reduce the increasing glutamic acid content in VTA-NAc-PFC neuroanatomical circuit and down-regulated NR2B expression, which may be one of mechanisms on reducing the effect of morphine CPP. C. yanhusuo preparations containing L-THP (1 x ) showed 24-fold effect of L-THP monomer of single application in terms of the behaviouristics of inhibitory effect on CPP as well as the similarity in terms of transmitter glutamic acid of in VTA-NAc-PFC neuroanatomical circuit and pharmacological mechanism of NR2B.

Animals ; Berberine Alkaloids ; therapeutic use ; Conditioning, Operant ; drug effects ; Corydalis ; chemistry ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Morphine ; antagonists & inhibitors ; Morphine Dependence ; drug therapy ; psychology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the clinical characteristics,outcome and prognostic factom of adolescent nasopharyngeal carcinoma.Methods Between Jan 1990 and Dec 2009,totally 148 pathological confirmed nasopharyngeal carcinoma(NPC)patients with age≤20 years were treated in our hospital,including stage Ⅱ 8,stage Ⅲ 58,stage Ⅳ 81,and unknown 1 when restaged by TNM system(UICC 2002),ninty-four(63.5%)patients were treated with radiotherapy alone,54(36.5%)patients were treated with radiotherapy combined with cisplatin-based chemotherapy.Results The median follow-up time for all patients was 44.5 months.The 5-year overall survival(OS),local-regional control(LRC)and distant metastasis-free survival(DMFS)rateswere 82.9%,85.1%and 78.6%.There were 42 patients(28.4%)failed with 16 regional recurrence and 29 distant metastasis,and 3 with both;bone metastasis was the most common site of distant metastasis(22/29).In univariate analysis,the adverse prognostic factors for OS were stage T4(X2=5.61,P=0.018),radiation dose<70 Gy(X2=5.30,P=0.021),for LRC was radiation dose<70 Gy and for distant metastasis-free survival was the disease history≥6month(X2=4.24,P=0.039).In multivariate analysis,radiation dose<70 Gy and stage T4 were the independent prognostic factors for OS(X2=5.73、5.56,P=0.017、0.018),for LRC was radiation dose<70 Gy(X2=5.81,P=0.016).Conclusions The outcome of the present series was excellent,total nagopharyngeal radiation dose less than 70 Gy is inappropriate.Reduce the distant metastasis and late toxicities were the future direction for the treatment of adolescent nasopharyngeal carcinoma.