[ ABSTRACT] AIM:To investigate the therapeutic effects of lentivirus-mediated shRNA targeting growth hormone secretagogue receptor 1a (GHSR1a) on colorectal cancer cell line SW480 both in vitro and in vivo.METHODS: Human GHSR1a sequence was used for the design of shRNA targeting GHSR1a, which was introduced to lentivirus, followed by transfection into SW480 cells.CCK-8 assay was performed to detect cell viability.The mRNA expression of GHSR1a and PTEN in colorectal cancer cells was detected by RT-PCR.The protein levels of GHSR1a, ghrelin, PTEN, p-AKT and p53 were determined by Western blot.Stable GHSR1a silencing in SW480 xenografts in nude mice was established.After the mice were sacrificed and weighted, immunohistochemistry was used to detect the positive expression of Ki-67 and PTEN in the tumors.RESULTS:GHSR1a was over-expressed in the malignant cells in comparison with the normal cells in vitro. The tumor specific lentivirus-mediated shRNA targeting GHSR1a gene and GHSR1a knockdown SW480 cells were success-fully constructed.After transfection with GHSR1a shRNA, the expression of GHSR1a at mRNA and protein levels was markedly inhibited in the SW480 cells.Furthermore, GHSR1a silencing by specific shRNA showed increased PTEN, inhi-bition of AKT phosphorylation and promotion of p53 release in the SW480 cells.In vivo results demonstrated that down-re-gulation of GHSR1a in the SW480 cells significantly decreased the expression of Ki-67 and increased the expression of PTEN in the tumor tissues, leading to a marked reduction in tumor weight in comparison to blank control or negative control
tumors.CONCLUSION:Down-regulation of GHSR1a by shRNA technique inhibits the growth of colorectal cancer cell line and xenograft tumor through activation of the PTEN/PI3K/AKT signaling pathways.

Objective To investigate the prevalence of hypertension and obesity and their main influencial factors among 6～12 years old children in Shenzhen.Method 1 140 children aged from 6 to 12 years old in 4 schools in Shenzhen were sampled by random cluster sampling,and their systolic blood pressure(SBP),diastolic blood pressure(DBP),body height,weight and other morphological parameters were measured.Results The prevalent rate of hypertension was 9.4%(8.6% for boys and 10.2% for girls);Rate of overweight and obesity for boys were 13.25% and 13.72,respectively,and the girl were 9.09% and 8.10%,respectively.There were a increasing trend toward SBP and DBP with age,especially SBP.After adjusted with age and gender,the partial correlation coefficients between BMI and SBP,DBP were 0.462 and 0.357,respectively(P

Objective To study the influence of aldehydes and tannic acid on soft capsules, approach how to prevent soft capsules from delayed disintegration. Methods Add aldehydes and tannic acid to soft capsules material ingredient, to proceed accelerating stability trial and test the disintegration time. Result Little increase of aldehydes amount had great effect on the disintegration time of soft capsules, different aldehydes and tannic acid had different effect on the disintegration time, adding some organic acid such as fumaric acid to the material would improve disintegrating property. Conclusion The amount of aldehydes and tannic acid contained in the material and ingredient should be controlled strictly when prepare soft capsules so that the disintegration time of capsules could be complying with the Pharmacopeia.

Objective To investigate the MRI features of pleomorphic xanthoastrocytoma (PXA).Methods 1 5 pathologically confirmed PXA cases were analyzed retrospectively.Clinical history and imaging features including location,size,shape,signal intensi-ty,enhancement and surrounding changes of those lesions were analyzed.Results All 1 5 cases were supratentorial and solitary le-sions,of which 9 lesions located in temporal lobe(60%).14 lesions contacted with the leptomeninges,and 1 lesion contacted with lat-eral ventricle wall.All lesions were solid-cystic,with different proportion of solid/cystic components.8 large lesions were predomi-nantly cystic(53.3%),3 small lesions were predominantly solid(20%),and 4 lesions had roughly equal cystic and solid proportions (26.7%).Solid components showed iso-intense or mild hypo-intense on T1 WI,iso-intense or mild hyper-intense on T2 WI,and signif-icant enhancement with contrast.Cyst fluid showed slightly hyper-intense in some cases.Cyst wall or septa enhancement was seen in 7 cases,and leptomeningeal enhancement was seen in 8 cases.Conclusion The MRI features of PXA are the characteristic of suprat-entorial solid-cystic lesions commonly seen in temporal lobe and contacting with leptomeninges.The typical features include “cyst with mural nodule”and “multiple cysts with irregular eccentric nodule”with significant enhancement of solid component and some cyst wall.MRI features of PXA is valuable in diagnosis and differential diagnosis of PXA.

Objective To verify the efficiency and stability of hydrogen-rich water preparation with hydrogen-rich rods. Methods ①Seven firenew hydrogen-rich rods were separately placed in seven plastic bottles, each filled with distilled water and soaked for 6 h, before the hydrogen concentration of the water was measured.This process was repeated 10 times.②After the hydrogen-rich rods with the strongest and weakest hydrogen product capacity were removed, the remaining 5 hydrogen-rich rods were placed separately into 5 plastic bottles filled with distilled water,put in a water bath pot at 20,40 and 60℃, respectively, and kept for 2, 4, 6, 8 and 10 h, respectively.Then, the hydrogen concentration, oxidation-reduction potential(ORP),and dissolved oxygen concentration(DO) were measured at various time points.③In order to determine the hydrogen emission rate from the hydrogen-rich water, the hydrogen-rich rods were constantly kept in some samples and the others were removed.All the sample bottle caps were kept open during the experimental process, and the hydrogen concentration was measured at such time points as 0, 10 and 30 min, 1, 2, 5, 12, 24, 30, 48 and 72 h, respectively.Results ①The hydrogen-rich rods used in this study could well meet the requirements.②When the environment temperature was kept constant, the hydrogen concentration of the water was increased with the soaking time of the hydrogen-rich rods, and the ORP of the water was reduced.However, the DO of the water was decreased with the rise of the environment temperature.③When the hydrogen-rich water was kept in opened plastic bottles with a 25 mm oral diameter, the hydrogen concentration of the samples with the hydrogen-rich rods reserved was almost about 0.50 ppm until 72 h, and that of the others was reduced to almost 0 ppm.Conclusion Our results demonstrate that the hydrogen-rich rods test is a simple and effective method for preparing hydrogen-rich water, which will be an valuable and useful method for using hydrogen-rich water in health promotion and prevention of chronic diseases.

Objective To analyze influence factors of serum A-TG level after DTC 131 I treatment ,to explore whether A-TG can be used as an indicator of follow-up ,recurrence and metastasis of DTC .Methods From 2008 January to 2013 February ,a total of 106 DTC patients underwent operation excisions of thyroid radioiodine were treated with 131 I .Before and 1 month after treatment , the levels of A-TG ,TG ,THS were measured .The relationship of A-TG and thyroid operation range operation times ,thyroid gland residual volume and time ,the levels of TSH and TG ,thyroid cancer metastasis and recurrence were examined .Results Serum A-TG concentration associated with operation scope ,frequency(P<0 .05) .It was showed that the A-TG concentration was positively correlated with the metastasis of thyroid cancer .Conclusion The serum TG level detection combined with A-TG and 131 I-WBS can improve the diagnostic sensitivity and accuracy of DTC recurrence and metastasis .

Objective To study the chemical constituents from Dipentodon sinicus. Methods Isolation and purification were carried out on silica gel column and Sephadex LH-20, the structures were elucidated on the basis of chemical evidences and spectral analysis. Results Nine compounds were isolated and their structures were identified as n-1-triacontanol (Ⅰ), ?-sitosterol (Ⅱ), pyrocatechol (Ⅲ), p-dihydroxyl benzene (Ⅳ), coniferyl aldehyde (Ⅴ), vanillic acid (Ⅵ), ergosterol peroxide (Ⅶ), ?-sitosterol-3-O-?-D-glucosides-3'-O-tritriacontanate (Ⅷ), and daucosterol (Ⅸ). Conclusion All the nine compounds are isolated from the title plant for the first time, among them, compound Ⅷ is a new compound named as dipentosinin.

Objective: To investigate the role of indoleamine 2, 3-dioxygenase in the development of uterine cervical squamous carcinoma. Methods: From January 2008 to December 2008, 116 uterine cervical carcinoma specimens and 18 metastatic lymph node specimens from patients with CIN Ⅰ-Ⅲ and uterine cervical squamous carcinoma were evaluated for iDO expression by immunohistochemistry. Twenty normal cervical specimens and 20 normal lymph node specimens were used as the controls. Results: The expression of IDO was not found in normal cervix and CIN Ⅰ. In CIN Ⅱ, IDO expres-sion was weakly positive in 2 cases (2/10, 20%) and negative in 8 cases (8/10, 80%). In CIN Ⅲ, IDO expression was weak-ly positive in 8 cases (8/13, 61.5%), positive in 1 case (1/13, 7.7%) and negative in 4 cases (4/13, 30.8%). The positive ex-pression rate of IDO in cervical cancer stage Ⅰ -Ⅳ was 100% (83/83). In cervical cancer stage Ⅰ A and Ⅰ B, the positive ex-pression rate of IDO was significantly higher than that in CIN Ⅱ and CIN Ⅲ (P<0.01). The positive expression rate of IDO in cervical cancer stage Ⅱ A-Ⅳ B was significantly higher than that in Ⅰ A and Ⅰ B. IDO expression was associated with cervi-cal cancer progression (OR=0.807, P<0.01). IDO expression in primary lesions with lymph node metastasis was significant-ly higher than that in those without lymph node metastasis. IDO expression rate was 100% in metastatic lymph nodes. The IDO expression was not associated with cervical squamous carcinoma differentiation degree (OR=-0.139,P>0.05). Conclu-sion: In CIN Ⅱ, escape mechanisms that stimulate cervical squamous carcinoma progression is gradually developed. IDO expression in metastatic lymph nodes is possibly associated with immune tolerance. IDO expression is not associated with differentiation degree of cervical squamous carcinoma. IDO may be a prognostic factor for uterine cervical squamous carci-noma and a therapeutic target for treatment.

Objective: α-asarone is a major effective component that can be isolated from Acorus tatarinowii Schott,a Chinese herbal medicine. Clinical investigations have shown that α-asarone has strong sedative and anti-convulsive action in the central nervous system. In recent years, several medicines containing a-asarone were applied in treatment of asthma, bronchitis, expectorant, or epilepsy. However, the underlying cellular mechanism of ct-asarone is still unknown. Here the authors considered EAAC1, the transporter for the excitatory glutamate, as a possible target. Methods: Supercritical CO2 fluid extraction and silica gel column chromatography were used to obtain ct-asarone from the rhizomes of Acorus tatarinowii Schott. Xenopus oocytes with heterologously expressed EAAC 1 were used as a model system. Rate of glutamate uptake was measured by means of isotopic tracer technique. Glutamate-induced current was recorded under two-electrode voltage clamp. 40μg/mL of ct-asarone was used for testing its effect on EAAC1 activity. Results: ct-asarone induced a slight, but still significant stimulation of rate of glutamate uptake by 15%. In contrast, EAACl-mediated current became reduced (by 30% at -100 mV). Since EAAC 1 can operate in transport and also in an ion-channel mode, the result indicates strong inhibition of the channel mode. This inhibition is voltage-dependent becoming larger at more negative potentials. Conclusion: The stimulation of glutamate uptake reduces glutamate concentration in the synaptic cleft and, hence, reduces excitatory synaptic activity. The inhibition on the ion-channel mode stabilizes the membrane potential, and therefore, also contributes to reduced excitatory activity.

Objective To investigate the differences between subjective and objective sleep quality in recurrent depressive patients and the correlative factors. Methods Thirty-two cases of recurrent depressive patients(the recurrent group) and 32 cases of first episode depressive patients(the control group)who met the inclusion were recruited in this study. The subjectivesleep quality ,mood and objective sleep quality of patients were estimated by Pittsburgh sleep quality index(PSQI),Hamilton anxiety Scale(HAMA),Hamilton depression Scale(HAMD),respectively. O ne whole-night polysomnography (PSG) was recorded. Results On subjec-tivesleep quality,the total sleep time was shorter,the sleep latency was longer,and the sleep efficiency was lower in the recurrent group compared with the control group. On objectivesleep quality ,the wake times were increased , the NREM1 phase was longer,the NREM2 phase was shorter,the RME phase was longer,the proportion of REM phase was increased,and the latency of REM phase was shorter in the recurrent group compared with the control group(P<0.05,respectively). There were significant differences between the subjective and objective sleep quali-ty in both groups,while the differences in the recurrent group were more significant(P < 0.01). In the recurrent group,thedifference of the total time between the subjective and objective sleep was positively correlated with PSQI score,the duration of disease and the number of attacks(P < 0.05),while,the differences of the sleep latency and sleep efficiency between the subjective and objective sleep were both positively correlated withPSQI score and the HAMA-14 score,respectively(P<0.05). Conclusion There was a significant difference between the subjec-tive and objective sleep quality of patients with recurrent depression ,and the differences were related to the course of disease,the number of attack,PSQI score and HAMA-14 score.