Objective To preparegraphene quantum dots (GQDs) and construct a novel biosensor for determination of dopamine (DA) and uric acid (UA).Methods The GQDs was prepared by carbonization method from citric acid as carbon sources.Differential pulse voltammetry was used by the modified electrodes to detect uric acid and dopamine,and the detection performance was evaluate.Collected three experimenters morning urine on July 29,2016.The proposed sensor was used for biological samples detection.Results The size of GQDs were between 3 to 5 nm,which was observed by transmission electron microscopy (TEM).The proposed sensor had good linear correlation of 50～600 μmol/L UA (r2 =0.996 6) and 2～ 240 μmol/L DA (r2 =0.992 5).In uric acid in urine samples' detection (n=3),RSD value was less than 3％.The standard addition recovery rates of UA and DA were in 95.7％～101.4％ and 96％～102％ respectively.Conclusion The biosensor based on GQDs for determination DA and UA had been constructed successfully.

Objective To explore the characteristics of defense mechanism in patients with obsessive-compulsive disorder(OCD).Methods Outpatients diagnosed as OCD were tested by Yale-Brown obsessive Compulsive Scale. Among them, 40 patients with score beyond or equal to 16 were selected as research group. 33 healthy individuals were selected as normal group. They were tested with Defense Style Questionnaire.Results ①The factor score of immature defense mechanisms in the OCD group is much higher than that in the normal group (P<0.01); the factor score of mature defense mechanisms in the OCD group is lower than that in the normal group (P<0.05); the factor score of middle defense mechanisms in the OCD group is much higher than that in the normal group (P<0.01). ②Passive aggression, acting out, splitting, regression, somatization in the immature defense mechanisms and relief, false altruism, isolation in the middle defense mechanisms are much more used by the OCD group than that in the normal group; but suppression and humor in the mature defense mechanisms are less used by the OCD group than that of the normal group(P<0.05 or P<0.01). ③Sublimation, suppression and consuming tendency are much more used in male patients than female ones in OCD group(P<0.05).Conclusion The patients with OCD tend to use the immature defense mechanisms and middle defense mechanisms, and there is some difference between male and female patients in using defense mechanisms.

ObjectiveTo investigate the association between the common variant rs1512268 single nucleotide polymorphism(SNP) of NKX3.1 gene and the risk of prostate cancer,and to explore its interaction with related risk factors.MethodsTotally 122 patients with prostate cancer and 105 age matched male people (prostatic specific antigen ＜ 4 μg/L,without family history of prostate cancer) as control group were enrolled.Polymerase chain reaction - high resolution melting curve(PCR - HRM) combined with gene sequencing methods were used to determine the distribution of allele and genotype frequencies of the rs1512268 SNP.ResultsThe distributions of GG,AG,AA genotypes were 42 cases(33.4％),66 cases(54.1％),14 cases(11.5％) in patients with prostate cancer,and 45 cases(42.9％),51 cases(48.6％),9 cases(8.6％) in healthy control,respectively.There were no significant differences in the distribution of genotype(x2 =1.70,0.69,0.52) and allele frequency (x2 =1.575) between the two groups(P＞ 0.05).The different genotypes of rs1512268 of NKX3.1 gene were not associated with age,Gleason score,PSA levels and clinical stage of prostate cancer (P＞0.05). Conclusions rs1512268 SNP of NKX3.1 gene is not obviously associated with prostate cancer and may be not the genetic risk factor in Chinese.

Objective To overcome the fact that SRV-NM virus can only multiple and amplify through partially pu-rified jaagsiekte retrovirus inoculated intratracheally in sheep but it cannot be augmented using in vitro cell culture, we con-structed JSRV-NM pseudovirions based on high efficiency packing system of lentivirus. Methods Lentivirus of three high efficiency packing plasmids system pMD.G, pCMV-HIV 8.2 and pHIV-eGFP was developed, and JSRV-NM-env coated plasmid pCMVJSRV-NM was used to substitute VSV-G virus coated plasmid pMD.G then co-transfected into 293T cells to replicate, package and produce restructured JSRV-NM pseudovirions. Gene expression of pseudovirion was determined through WPRE using real time PCR; Virus infectivity was detected through inoculating JSRV-NM pseudovirions into 24 pore plates. Results We construct JSRV-NM pseudovirions successfully based on the lentivirus system. JSRV-NM pseudo-virions can also be concentrated to higher titer (108 TU/mL detected by real time PCR by ultracentrifugation without signifi-cant loss of activity. JSRV-NM and VSV-G pseudovirions infected on Hela cells (both MOI= 3) respectively and no obvi-ous difference were shown on their infection efficiency detected by real time PCR. Conclusion Based on lentivirus system, JSRV-NM pseudovirions can be multipled and amplified in 293T cell culture in vitro. JSRV-NM pseudovirions is stable without loss its infection activity and the requirements of biological laboratory safety II was also met. JSRV-NM pseudoviri-ons will provide a useful tool for further study of JSRV-NM-env infection across species or its induction of lung adenocarci-noma.

Objective:To investigate the impact of aging on the correlation between the intestinal microorganism Akkermansia and obesity, and to analyze the features of the correlation in the elderly population. Methods:This was a cross-sectional study.A total of 6896 cases were collected from the Guangdong intestinal microbiome in 2018, aged 18-94 years old, including 3806 females, 1641 cases with abdominal obesity(23.7%)and 707 cases with systemic obesity(10.3%). The 16S rRNA sequencing data were from individuals of Cantonese descent.The abundance of Akkermansia was calculated after data cleaning, clustering and annotation.The type of abdominal obesity or systemic obesity was diagnosed based on the standards of the Working Group on Obesity in China(2002). According to the five quintiles of the abundance of Akkermansia, subjects were divided into Q1~Q5(Q1-Q4: n=1379, Q5: n=1380). Logistic regression was used to study the relationship between Akkermansia and obesity after adjusting for common confoundors such as gender.Subjects were subgrouped into two types of age groups: the <65 group(n=5467)and the ≥65 group(n=1519); the <70 group(n=6136)and the ≥70 group(n=850). Age windows were used to analyze changes in characteristics of this relationship with increasing age. Results:There were significant differences in age and gender among different Akkermansia groups( t/ χ2=3.51, -5.03, P<0.01). Logistic regression analysis showed that after adjusting for two main confounding factors, age and gender, the risk of systemic obesity and abdominal obesity gradually decreased from Q2 to Q5 group, compared with Q1 group( P<0.001). The correlation between Akkermansia and obesity decreased with age.The protective effect of Akkermansia on obesity was weaker in the ≥65 and ≥70 groups, respectively, than in the <65 and <70 groups. Conclusions:Akkermansia is a protective factor for obesity, but the protective effect is affected by aging and weakened in the elderly.

Seizure-sensitive Fisher 344 rats were prepared 7 d after seizure episodes induced by a single subcutaneous administration of convulsive dose (7.5 mg/kg)of kainic acid. The activator protein-1 (AP-1) DNA binding activity and its components in the hippocampus of such rats were examined by Gel shift,Super-shift and Western blot assay. Gel shift assay showed that the basal level of AP-1 DNA binding activity was significantly higher than that of control rat hippocampus. Super-shift showed that only Fra and Jun D antibodies could super-shift AP-1 complex to a higher position. It suggested that the AP-1 complex was composed of Fra and Jun D in hippocampus of the seizure-sensitive rats. Western blot analysis further confirmed that Jun D proteins contained components of 43,39 and 28 kDa. When secondary seizure stimulations were given to seizure-sensitive rats, the AP-1 DNA binding activity was attenuated and the components of AP-1proteins changed transiently. In light of our results and those of literature, it appears that all the mentioned changes may play an important role in induction of long-lasting enhancement in seizure susceptibility.

Objective To study Epstein-Barr vires infection and p16 protein abnormal expresson in carcinogenesis and progression of gastric adenocarcinomas (GAC). Methods Immunohistochemical staining SP method was used to detect the expression of LMP-1 and p16 in 97 cases of GAC. Results EBV LMP-1 and p16 protein were detected in 30.9% (30/97) and in 63.91% (62/97) cases of gastric adenocarcinomas respectively.There was no significant difference between EBV-positive and EBV-negative gastric carcinomas in sex, histologic type, depth of tumor invision, lymph node metastasis and clinical stages (P>0.05);overexpression of p16 was associated with lymph node metastasis and clinical stages; no correlation was found between the expression of EBV LMP-1 and p16 protein. Conclusion ①EBV play a role in carcinogensis of GAC. ②P16 gene abnormality is frequently involved in GAC and might be one of the important prognostic factors. ③EBV infection and p16 alteration are two independent roles in GAC carcinogenesis.

OBJECTIVETo observe the effects of acupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) on distribution taxis of paclitaxel in mice with lung cancer to discuss targeted relationship between acupoints and corresponding viscera.

METHODSAccording to randomized digital table, 315 SPF-grade BALB/C female mice were divided into 7 groups: blank group (group A), model group (group B), medication group (group C), acupuncture at non-acupoint group (group D), acupuncture at Feishu group (group E), acupuncture at Lingtai group (group F) and acupuncture at Feishu and Lingtai group (group G), 45 mice in each one. Except the blank group, the remaining groups were treated with N-nitroso-tris-chloroethyl urea (NTCU) to establish the model of squamous-cell carcinoma. After model establishment, group A, group B and group C were not treated with acupuncture; group A and group B were treated with intraperitoneal injection of 0.9% sodium chlorvde solution by 6 mL/kg while group C was treated with intraperitoneal injection of paclitaxel by 6 mL/kg. The group D, group E, group F and group G were treated with acupuncture at non-acupoint, "Feishu" (BL 13), "Lingtai" (GV 10) and "Feishu" (BL 13) plus "Lingtai" (GV 10), respectively, then were intraperitoneally injected with paclitaxel by 6 mL/kg. The treatment was all given once a day for continuous 10 days. 15 min, 30 min, 1 h, 2 h, 8 h, 12 h and 24 h after the treatments, 6 mice in each group were randomly selected and sacrificed to collect samples of lung, liver, spleen, kidney and heart, etc. High performance liquid chromatography was applied to measure the concentration of paclitaxel in each organ (lung, liver, spleen, kidney and heart) at different time points.

RESULTS(1) The content of paclitaxel in lung, kidney and heart reached the peak at 2 h, then decreased significantly in group C, group D, group E, group F and group G; the content of paclitaxel in spleen showed downtrend at each time point. The content of paclitaxel in liver reached the peak at 2 h in group C and group D; the content of paclitaxel reached the peak at 8 h in group E, group F and group G. (2) The content of paclitaxel in lung in group E and group G was higher than that in group C and group D at each time point (all P < 0.01); the content of paclitaxel in lung in group F was higher than that in group C (P < 0.01) and group D (P < 0.01) only at time point of 2 h. The content of paclitaxel in lung in group G was higher than that in group F at each time point (all P < 0.05). There was no statistical difference between group G and group E (all P > 0.05).

CONCLUSIONAcupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) could influ- ence the metabolism of paclitaxel in lung-cancer mice, leading to distribution change in each organ. As a result, it could cause targeting effects, which is more significant at "Feishu" (BL 13) and "Lingtai" (GV 10).

Acupuncture Points ; Acupuncture Therapy ; Animals ; Antineoplastic Agents, Phytogenic ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Female ; Humans ; Lung Neoplasms ; drug therapy ; therapy ; Mice ; Mice, Inbred BALB C ; Paclitaxel ; pharmacokinetics ; Taxus ; chemistry

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

OBJECTIVETo evaluate the clinical efficacy of the Chinese medicine Yiqigushen Capsule combined with levofloxacin on type III A prostatitis.

METHODSA total of 180 patients with type III A prostatitis were equally randomized into a treatment group, which received Yiqigushen Capsule combined with levofloxacin, and a control group, which was given Yiqigushen Capsule only, both for 4 weeks. NIH-CPSI scores and leukocyte counts in the expressed prostatic secretions of all the patients were analyzed before and after the treatment.

RESULTSAfter the treatment, NIH-CPSI scores and leukocyte counts in the expressed prostatic secretions of both the groups were significantly decreased compared with pre-treatment (P < 0.05 or 0.01), with statistic differences in NIH-CPSI scores but not in leukocyte counts (P < 0.05) between the two groups.

CONCLUSIONYiqigushen Capsule combined with levofloxacin is highly effective for type III A prostatitis.

Adult ; Anti-Infective Agents, Urinary ; therapeutic use ; Capsules ; Chronic Disease ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Levofloxacin ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ofloxacin ; therapeutic use ; Prostatitis ; drug therapy ; pathology ; Treatment Outcome