OBJECTIVETo study the effect of aqueous extract of Taxus chinensis var. mairei (AETC) combined Erlotnib on the growth of A549 xenograft in nude mice and its mechanism.

METHODSThe xenograft model in nude mice was established by inoculating A549 cells subcutaneously. BALB/c nude mice bearing A549 xenograft were randomly divided into six groups, i.e., the low dose Erlotinib group (A) , the standard dose Erlotnib group (B) , the low dose Erlotinib combined AETC group (C), the standard dose Erlotnib combined AETC group (D), the AETC group (E), the control group (F), 12 in each group. Different medication was performed for 7 successive weeks after 24 h. One mL blood was withdrawn and tumor tissues taken. The tumor inhibition rate was calculated. The combined effect was analyzed by Jin's Formula [Q = Ea + b/(Ea + Eb-Ea x Eb) ]. mRNA and protein expression levels of epidermal growth factor receptor (EGFR), cyclooxygenase-2 (COX-2), and B cell lymphoma-2 (Bcl-2) in xenografts were detected using real-time RT-PCR and ELISA.

RESULTSCompared with Group F, the xenograft weight was obviously lowered in Group B-E (P < 0.05, P < 0.01). The q value was 0.92 in Group C and 0.96 in Group D, which was obtained by simple adding of the two drugs. Compared with Group F, EG- FR mRNA expression in Group D and E, COX-2 mRNA expression in Group A-E; Bcl-2 mRNA expression in Group B-D; COX-2 protein expression in Group B-E; Bcl-2 protein expression in Group C and D were obviously lowered with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONSAETC combined low dose and standard dose Erlotinib had synergistic effect on tumor inhibition. Its mechanism might be associated with down-regulating mRNA and protein expression levels of COX-2 and Bcl-2.

Animals ; Antineoplastic Agents ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cyclooxygenase 2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Erlotinib Hydrochloride ; pharmacology ; Heterografts ; Lung Neoplasms ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Receptor, Epidermal Growth Factor ; metabolism ; Taxus ; Transplantation, Heterologous

This study is to investigate the anti-tumor activities of a novel cyclophosphamide derivate 4, 6-diphenyl cyclophosphamide (9b) in vivo and in vitro, and its possible mechanism of action. The inhibitory effects of 9b on human hepatoma cell line HepG2, human breast carcinoma cell line MCF-7 and human myeloid leukemia cell line K562 were measured by MTT assay in vitro. Cell cycle distribution and apoptotic rate were evaluated by flow cytometry. To evaluate the anti-tumor effect of 9b in vivo, mouse model bearing inoculated H22 tumor was established. The results indicated that 9b could inhibit the proliferation of HepG2, MCF-7 and K562 cells in a dose and time dependent manner. The ICo50 values of 9b were 32.34 micromol.L-1 to HepG2 cells, 87.07 micromol.L-1 to MCF-7 cells and 149.10 micromol.L-1 to K562 cells after incubation for 48 h. The results of flow cytometry indicated that after being treated for 48 h with different concentrations of 9b, the ratios of HepG2, MCF-7 cells at the Go/G1 phase and K562 cells at the G0/Gl phase and G2/M phase increased significantly compared with control group, and the apoptotic rate increased with the increase of the concentration of 9b. 9b could significantly reduce tumor weight of H22 solid tumor mouse model in vivo. To summarize, 9b showed significantly anti-tumor activity in vivo and in vitro, of which the mechanism might be associated with the change of cell cycle distribution and induction of tumor cell apoptosis.
Animals ; Antineoplastic Agents, Alkylating ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclophosphamide ; analogs & derivatives ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms, Experimental ; pathology ; Male ; Mice ; Molecular Structure ; Random Allocation ; Tumor Burden ; drug effects

OBJECTIVESTo examine the Epstein-Barr virus (EBV) in primary lung carcinoma tissue, and to investigate the relationship between EBV infection and tumorigenesis of lung cancer.

METHODSFormalin-fixed and paraffin-embedded lung tissue specimens from surgically resected lung carcinoma tissues of 108 cases treated in Tanshan area from 2001 to 2006, which were confirmed further by histopathological examination after hematoxylin-eosin (HE) staining, were used to observe the EBV encoded RNA-1 (EBER1) using in situ hybridization (ISH).

RESULTSEBER1 was detected in 36 of the 108 primary lung carcinoma cases, and in 1 of the 22 normal lung tissues. The positive rates of EBV infection in squamous cell carcinoma, adenocarcinoma, small cell carcinoma and large cell carcinoma were 35.9%, 31.6% 31.0%, 1/2, respectively. Gender, age and clinicohistopathological type were not found to have any correlation with EBER1 expression, but EBER1 expression in groups of cases with poorly and moderately differentiated carcinomas was significantly higher than those in the group of cases with well differentiated carcinoma, and the EBER1 expression in the right lung was higher than in the left lung.

CONCLUSIONSThe frequency of EBV infection in this series of patients from Tangshan area was 33.3%, the results suggest that there is a relationship between EBV infection and the occurrence of the primary lung carcinoma, EBV infection might be one of the potential causes to induce lung cancer.

Epstein-Barr Virus Infections ; diagnosis ; virology ; Herpesvirus 4, Human ; genetics ; Humans ; In Situ Hybridization ; methods ; Lung Neoplasms ; pathology ; virology ; RNA, Viral ; genetics

Objective:By applying moxibustion to the eight confluent points in different periods of time,to observe the changes in thermal pain threshold latency of acupoints based on Fei Teng Ba Fa.Methods:A total of 468 healthy college student volunteers received moxibustion at the eight confluent points in three different periods of time,i.e.Chen (7:00-9:00),Wu (11:00-13:00) and Xu (19:00-21:00).The thermal pain threshold latency was adopted to measure the changes in pain threshold of the eight confluent points under different conditions (different periods of time,different genders,different acupoints and different states of the acupoints) based on Fei Teng Ba Fa.Results:Finally,thirty subjects dropped out and 438 subjects were included.The comparison of thermal pain threshold latencies of the eight confluent points in the same opening or closing state based on Fei Teng Ba Fa:latencies of the closing points and adjunct points were significantly different in different periods of time (P＜0.05);the latencies of the males were significantly longer than those of the females (P＜0.05);there was no significant difference in the latency between the left and right sides (P＞0.05);in the female group,there was a significant difference in the latency between the lower-limb points and the upper-limb points (P＜0.05).The comparison of thermal point threshold latencies of the eight confluent points in different opening or closing state:in the period of Wu (11:00-13:00),the latencies of the opening points were significantly longer than those of the closing points and adjunct points (P＜0.05);for men,their opening and closing points had significantly longer thermal pain threshold latencies than their adjunct points (P＜0.05);despite the gender,the latencies of the upper limb opening and closing points were significantly longer than the latency of the adjunct points (P＜0.05);in the female group,the latencies of the lower-limb opening points were significantly shorter than those of the lower-limb closing and adjunct points (P＜0.05).Conclusion:Based on Fei Teng Ba Fa,the pain thresholds of the eight confluent points vary in different periods of time,gender,acupoint location and opening/closing state,which can be taken as the evidence of making time-based acupuncture-moxibustion prescriptions.

Objective To investigate the effect of excessive fluoride,aluminum on Zn,Fe,Ca,Mg,Cuin rat blood.Methods Forty eight SD rats were randomly divided into 4 groups matched with their weights:control group,high aluminum group,high fluorine group and high fluorine-aluminum group.Aluminum content in their drinking water was 0,90,0,90 mg/L respectively.Fluorine content of their feed was 5.2,5.2,106.0,106.0 mg/kg and aluminum Was 6.8,6.8,19.7,19.7 mg/kg respectively.90 days later,the level of blood Zn,Fe,Ca,Mg, Cu Was detected by the atomic absorption spectrometry.Results Compared among these groups,Zn,Fe,Mg and Cu content of the whole blood had significant difierences(F=46.25,14.74,6.10,2.93,P<0.05),while Ca content of the whole blood did not significantly change(F=2.81.P>0.05).Factorial analysis showed that excessive intake of aluminum could significantly decreased Zn,Fe,Mg content of the blood(F=42.66,5.41,7.04,P<0.05)and excessive intake of fluorine could significantly decreased Zn,Fe,Mg,Cu content of the blood(F=64.50,37.90,9.75,6.74, P<0.05).The coexistence offluorine and Muminum had interaction to the level of Zn(F=31.59,P<0.05)and did not obviously interact with other elements(F=0.91,1.63,1.51.0.00,P>0.05).Compared with the control group [(131.30 ±13.86)μmol/L,(10.24 ±1.02),(1.71 ±0.19)mmol/L,(20.43 ±4.42)μmol/L],Zn content in the high aluminum group[(90.84±9.98)μmol/L]decreased significantly(P<0.05),so did Zn,Fe,Mg content in tlle high fluorine group[(85.85 ±10.92)μmol/L,(8.49 ±0.68),(1.52 ±0.13)mmol/L],the difference being statistically significant(P<0.05)0Zn,Fe,Mg,Cu content in the high fluorine-aluminum group,being(82.82 ±11.00)μmol/L, (8.16±0.45),(1.46±0.09)mmoL/L,(15.69±2.38)μmol/L,respectively,all decreased signitlcarIdy(P<0.05). Compared with the high aluminum group[(9.43±1.09)mmol/L],Fe content of the high fluorine aluminum group[(8.16±0.45)mmol/L]decreased significantly(P<0.05).Conclusions Excessive fluoride can cause blood zn, Fe,Mg,Cu decline,so can excessive aluminum.Combination of excessive fluofine and aiuminum has 8ignificant synergic effect on the level of Zn but have rio influence on Ca.

Objective To investigate the Epstein-Barr virus (EBV) infection, the expression of EBV latent membrane protein 1 ( LMP1 ) and oncogene bcl-2 in lung cancer patients. Methods EBER1 in 108 cases of lung cancer were detected with in situ hybridization. EBV positive and negative lung cancer tissues were analysed for the expression of LMP1 and Bcl-2 by immnohistochemistry. The average area (AA)and integral optical density (IA) of each sample was measured with the digital medical image analyzing system. ResultsIn 108 cases of lung cancer, 36 cases were EBER1 positive and 7 cases were LMP1positive. The expression of Bcl-2 was higher in EBV positive lung cancer tissues than that in EBV negative.The AA value was 58014. 23 ±6918.45 and 38156.22 ± 4096. 79, while the IA value was 11. 00 ± 1.48 and 8.03 ±0. 78 respectively. No statistic difference was fund in the expression of Bcl-2 betwen LMP1 positive and negative lung cancer tisssues. ConclusionEBV infection in lung cancer increased the expression of bcl-2, which may play a role in the occurrence or development of lung cancer. The increased expression of Bcl-2 may not be induced by LMP1. The exact mechanism need further study.

OBJECTIVETo observe the effect of needling "Housanli" (ST 36) with different retaining-needle time on the pain threshold of mice using the hot water tail-flick test, and investigate the relationship between the retaining-needle time and the therapeutic effect.

METHODSTwenty-four healthy mice were randomly divided into four groups: no retaining-needle group (group A), retaining-needle 10 min group (group B), retaining-needle 20 min group (group C) and retaining-needle 30 min group (group D), with 6 mice for each group. After acupuncture "Housanli", the tail of mouse was put into the hot water at 50 degrees C, and the intervening time from the tail entered water to the tail flicked out of water was recorded and analyzed for each group.

RESULTSAfter acupuncturing "Housanli", the pain threshold was significantly improved with the hot water tail-flick test in both group A and group C (both P<0.01), however, there was no significant difference in both group B and group D (both P>0.05). Within 30 min of retaining-needle, the effect curve of acupuncture analgesia was showed in a waved line. In contrast, the maximum value of latent period on the tail-flick was detected in the group C that was the biggest effect of acupuncture analgesia among the four groups.

CONCLUSIONThe best retaining-needle time of hand-acupuncture for acupuncture analgesia is 20 min.

Acupuncture Analgesia ; methods ; Acupuncture Points ; Animals ; Male ; Mice ; Needles ; Pain ; physiopathology ; Pain Management ; Pain Measurement ; Pain Threshold ; Random Allocation ; Time Factors

OBJECTIVETo compare the difference in the efficacy on gait time cycle of ischemic stroke between yin-yang respiratory reinforcing and reducing needling technique (yin-yang needling) and the conventional acupuncture.

METHODSSixty cases of ischemic stroke were randomized into a conventional acupuncture group and a yin-yang needling group, 30 cases in each one. The basic treatment (the control of blood pressure, blood sugar and blood lipid, the intravenous drops of ginkgo leaf extract and dipyridamole injection and vinpocetine injection) were applied in the two groups. Additionally, in the conventional acupuncture group, the acupoints of the Stomach Meridian of Foot-Yangming [Biguan (ST 31), Liangqiu (ST 34), Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), etc.] were selected and stimulated with the even needling technique. In the yin-yang needling group, the acupoints of yin meridians such as Zuwuli (LR 10), Xuehai (SP 10), Yinlingquan (SP 9) and Sanyinjiao (SP 6), etc. and the acupoints of yang meridians such as Biguan (ST 31), Liangqiu (ST 34) and Yanglingquan (GB 34), etc. were selected. The reducing manipulation of respiratory reinforcing and reducing technique was applied to the acupoints of yin meridians and the reinforcing manipulation was applied to the acupoints of yang meridians. The kinematics time parameters were determined and compared before and 4 weeks after treatment.

RESULTSAfter treatment, the differences in the gait cycle, the phase time of standing (%), the phase time of single support (%), the phase time of unilateral sway (%) on the affected (healthy) foot and phase time of double support (%) were significant as compared with those before treatment in the patients of the two groups (all P < 0.05), in which, the gait cycle (1.75 +/- 0.21 vs 2.02 +/- 0.37), the phase time of standing (%) on the affected (healthy) foot [(65.41 +/- 5.20)% vs (68.37 +/- 6.24)%, (70.99 +/- 6.47)% vs (74.51 +/- 5.19)%], the phase time of unilateral sway (%) on the affected (healthy) foot [(36.08 +/- 4.86)% vs (33.65 +/- 2.94)%, (31.04 +/- 3.41)% vs (26.77 +/- 2.67)%] and the phase time of double support (%) [(36.91 +/- 5.10)% vs (41.22 +/- 5.39)%] in the yin-yang needling group were improved much obviously after treatment as compared with those in the conventional acupuncture group. The differences in support phase time (%), single support phase time (%) and sway phase time (%) were significant between the affected limb and healthy limb of the two groups after treatment (all P < 0.05).

CONCLUSIONYin-yang respiratory reinforcing and reducing needling technique effectively improves hemiplegic gait movement cycle and walking function in patients of ischemic stroke, which is superior to the conventional acupuncture treatment.

Acupuncture Therapy ; instrumentation ; methods ; Aged ; Female ; Gait ; Humans ; Hypoxia-Ischemia, Brain ; physiopathology ; therapy ; Male ; Meridians ; Middle Aged ; Stroke ; physiopathology ; therapy ; Time Factors ; Yin-Yang

Objective To classify and grade the environments with Oncomelania hupensis snails inside embankment in marsh-land and lake areas,so as to improve the work efficiency and realize the systematic management. Methods The schistosomiasis endemic area Liaodi and Xiongdi villages in Jiangling County,Hubei Province were selected as the experiment villages,and the environments with snails in the two villages were divided into sections with a length of 500m,then the snail situation were sur-veyed by the systematic sampling method with 10m and 50m a frame respectively. The environments were classified according to the discovery of infected snails and wild fecal contamination,and the numbers of sites with snails and their areas in different class-es were recorded. Meanwhile,the data of sites with infected snails in Jingzhou City during the recent 5 years were collected and graded according to the average density of snails,so as to discuss the correlativity between the grades of environments with snails and the numbers of sites with infected snails and the occurrence rate of frames with snails. Results There were 1 967 sites with in-fected snails in Jingzhou City during the recent 5 years,and there was a positive correlation between the grades of environments with snails and the occurrence rate of frames with snails(rs=0.77,P<0.01),when the grade was 5,the constituent ratio of num-bers of sites with infected snails was 3.1%,and when the grade was 2-3,the ratio raised to 56.3%. There were 39 sites with snails in the two villages,the Class 1,2 and 3 environments included 1,18 and 20 sites,with the areas of 1 080,51 640 m2 and 41 220 m2 , respectively. There were no statistically significant difference between the grades divided by snail survey with 10m and 50m a frame(χ2=4.667,P>0.05),but the time-consuming of the former was 3 times of the latter. Conclusions The classification and grading of the environments with snails inside embankment in marshland and lake areas can master the key point of the snail survey. The subsection and setting frames at a suitable distance can save time and manpower,improve work efficiency,as well as understand the distribution of snail status of the environment with snails,which can realize the sort management of the snail envi-ronments inside embankment.

OBJECTIVETo investigate genomic DNA imbalances in ependymomas (EDMs) and their correlations with the tumor histological types, grades, locations, patients' gender and age.

METHODSChromosomal gains and losses in 16 cases of EDM were analyzed using comparative genomic hybridization.

RESULTSChromosomal regional gain and loss were found in 15 and 13 of 16 EDM cases respectively including totally 24 regional gains and 19 regional losses in all the tumors studied. Both regional gains and losses were mostly seen in myxopapillary EDMs (MPE, WHO grade I), more commonly seen in cellular EDMs (CE, WHO grade II) and tanycytic EDMs (TE, WHO grade II) than in anaplastic EDMs (AE, WHO grade III). Some of the regional gains and losses appeared only in one subtype of MPE, CE, TE and AE cases resulting in development of specific imbalance profiles of certain subtype in these cases. MPE, CE and TE often had +7. Chromosomal +5 occurred only in MPE and CE, and -22q was only seen in CE and TE. AE frequently had +1q, but none had +5, +7, -4q, -19q and -22q. The frequencies of any regional gain or loss were not affected by patients' genders (P > 0.05). Chromosomal +1q and +7p happened predominantly in intracranial EDMs with an averagely onset age of

CONCLUSIONSThe frequencies of chromosomal imbalances in EDMs decrease as the tumor grade increases. Characteristic chromosomal imbalances in each subtype may play an important role in determination of histological phenotypes and tumor grades. Chromosomal +1q, +5, +7p, +7, -4q, -19q and -22q are alterations which may dictate the biological behaviors of these tumors and the patients' prognosis.

Adolescent ; Adult ; Aged ; Brain Neoplasms ; classification ; genetics ; pathology ; Child ; Chromosome Aberrations ; Comparative Genomic Hybridization ; DNA, Neoplasm ; genetics ; Ependymoma ; classification ; genetics ; pathology ; Female ; Humans ; Male ; Middle Aged ; Spinal Cord Neoplasms ; classification ; genetics ; pathology ; Young Adult