Object In order to identify the resources of official and unofficial plants in Rheum L.at molecular level, a random amplified polymorphic DNA (RAPD) analysis was performed. Methods The amplification program of polymerase chain reaction (PCR) was optimized. Arbitrary decamer primers of 121 based on the individual selecting method, and 64 based on the BSA method were screened. Results Four primers among all screened primers were found to be useful to amplify specific strips to the official plants of Rheum L. Conclusion This method is reliable, accurate, quick and reproducible for authenticating the official and unofficial plants of Rheum L. at molecular level. Moreover, the experiment results indicate that it is more effective to screen the primers using BSA method than using individual selecting method.

ObjectiveTo establish an analytical method with high sensitivity and wide range in biotin-avidin and time-resolved fluoroimmunoassay system for the quantitative detection of antinuclear antibody (ANA)-Ig (GAM).MethodsANA in standard preparation or sample was combined with solid nuclear antigen,biotinylation antibody and the europium(Eu3+)-labelled avidin to form the compounds of solid nuclear antigen-antibody-biotinylation antibody-SA-Eu3+.The fluorescence enhancement fluid was added to dis-sociate the Eu3+,the content of ANA was directly proportional to fluorescence intensity,the BAS-TRFLA was established for the quantitative detection of ANA-Ig (GAM).The sensitivity,specificity,reliability and range of detection were evaluated.Semm of 50 blood donor,105 patients with systemic lupus erythematosus (SLE),109 patients with rheumatoid arthritis(RA),25 patients with PBC,29 patients with Sj(o)gren's syndrome (SS),23 patients with scleroderma,25 patients with MCTD were included in this study.The positive rate was calculated.ResultsThe inner-group difference between high,medium and low densities mixture serum was 3.13％,3.74％ and 5.76％ and the inter-group precision rate was 5.31％,6.25％ and 7.46％ in BAS-TRFLA.The sensitivity of TRFIA was better than that of the ELISA method.The low detection limit was 2.24 U/ml.The mean recovery rate was 100.74％.The results measured by the TRFIA and ELISA methods were closely correlated(R2=0.978).The positive rate of blood donor,and patient with SLE,RA,PBC,SS,scleroderma and MCTD were 0,100％,23％,96％,38％,91％,92％ respectively.When compared with ELISA,the detection range of TRFIA was wider,and stability was better.ConclusionBAS-TRFIA is a stable method for detection of ANA with high sensitive and wide range of detection.It is important for the early diagnosis of autoimmune disease and monitoring the treatment efficacy of autoimmune disease.And this method may be widely used in clinical laboratories in the future.

Objective: In order to establish an effectively analytic method about the content of lead, arsenic, cadmium, mercury and copper. Methods:According to the standard addition method, matrix modifiers (nickle nitrate and diammonium orthphosphate) were added to sample and the contents of Pb, As, Cd and Cu were determined by graphite furnace atomic absorption spectrometry, reduce agent (acid tin bichloride was added to sample and the content of Hg was determined by cold vapour atomic absorption method. Results: The average recoveries of lead arsenic, cadmium, copper and mercury were Pb=97.5%, RSD = 2.75%; As=96.6%, RSD = 2.23%; Cd= 98.2% , RSD = 1.32% ;Cu=97.8%, RSD =1.19%;Hg= 98.8% , RSD = 1.85 , respectively. Conclusion: The method is sensible, accurate and reproducible. and applied for quality control of content of heavy metals in Qijidehuang Pills.

Objective To improve the quality of aiye processed products, an eucalyptol content in commercially available aiye two processed products of chao aiye and aiye tan was investigated.Methods A capillary gas chromatography was used.The sample was prepared with n-hexane by reflux condensation.Chromatographic conditions: The separation was carried on an Ailgent DB-1 capillary column(30 mm ×0.320 mm ×0.25 μm). Inlet temperature was 200℃ and FID temperature was 250℃.The programmed column temperature was set as follows:maintained at 100℃ for 6 min and raised to 160℃ at the rate of 20℃/min followed by holding for 3min.The splitting-ratio was 5.0:1.The carried gas was nitrogen, flow rate was 1.0 mL/min.Injection volume was 1μL.Results In the given chromatographic conditions, the eucalyptol chromatographic separation had good, and the separation degree was greater than 1.5 between eucalyptol and other impurity peak.The linear range of eucalyptol was 11.4-114.0 mg/mL(r=0.999 5). Methods repeatability and recovery were good.The minimum limit of quantification was 0.5μg/mL.The results of determination of eucalyptol show that the eucalyptol content in the commercially available 11 batch of chao aiye was between 5.6-78.2 μg /g, and 12 batch of aiye tan had no eucalyptol. Conclusion The processing technology of current commercially available aiye processed products of chao aiye and aiye tan need to be improved, and the quality standard need to be improved.

Objective To explore the effect of reconstructing unilateral cleft lip by changing the arc-shaped incision, combined with the 3D reconstruction of upper lip muscles.Methods Twenty unilateral cleft lip patients were treated by using a new surgical operation, the 3D reconstruction of upper lip muscle, to restore normal anatomy and stress of the mucous membrane, muscle and skin.Operation scar was designed for straight line, located on the philtral ridges of the contour line;phitrum and philtral ridges were rebuilt, and postoperative scar reduced.Results A lot of 20 patients had no local infection, hemorrhage, complex crack, and were stage I incision healing.Followed up for 1-8 months postoperatively, the patient's lip bow line continuity was good, with symmetrical shape and good phitrum and philtral ridges;scar was hidden on the philtral ridges of the contour line, and no obvious upper lip scar contracture found through the follow-up period.Conclusions This improved method is simple in the incision design, and less scar hidden on the philtral ridges of the contour line after operation, which can maximize the recovery of the appearance of nose and upper lip with satisfactory effect.It is a feasible improvement method of repairing unilateral cleft lip.

OBJECTIVETo explore the expression pattern of Mus81 gene in Chinese Han patients with colorectal cancer and analyze its correlation with the clinicopathological parameters and the tumor markers.

METHODSMus81 expression was detected using nested quantitative real-time PCR in the colorectal cancer tissues and corresponding adjacent normal tissues from 43 Chinese Han patients. The correlations of Mus81 expression profile with the clinicopathological parameters and common tumor markers were evaluated.

RESULTSMus81 expression level was significantly lower in the colorectal cancer tissues than in the corresponding adjacent normal tissues (114.6 ± 68.0 vs 202.5 ± 109.0, P<0.001). Of the 43 patients, 32 (74.4%) showed down-regulated Mus81 expression, which correlated significantly with distant metastasis (P=0.043), high TNM stage (P=0.022) and high P53 protein expression (P=0.011) of the tumor.

CONCLUSIONDown-regulation of Mus81 in colorectal cancer is correlated with tumor metastasis and progression, suggesting the value of Mus81 as a potential marker for colorectal cancer in Chinese Han patients.

Biomarkers, Tumor ; Colorectal Neoplasms ; diagnosis ; genetics ; DNA-Binding Proteins ; genetics ; Disease Progression ; Down-Regulation ; Endonucleases ; genetics ; Humans ; Prognosis ; Real-Time Polymerase Chain Reaction

Background and objective Lung cancer is one of the most serious disease and the incidence of non-small cell lung cancer (NSCLC) is the highest in lung cancer. The main reason for the failure of chemotherapy is the tolerance to cisplatin. Transcriptional regulator SOX4 plays an important role in the occurrence and development of many tumors, and regulates Wnt signaling pathway by regulating the expression of β-catenin. We aimed to investigate the role of SOX4 on cisplat-in-resistance in NSCLC cell A549 cell. Methods The cisplatin-resistance lung cancer cell line A549/DDP was constructed by induction method in vitro, and cisplatin-resistance detected by CCK8 assay. Growth curves of A549 and A549/DDP was cal-culated. The expression level of SOX4 in A549 and A549/DDP cells were detected by Western blot. A549/DDP were knock-down of SOX4 by siRNA transfection, and the cisplatin-resistance of detected by CCK-8 assay, the expression level of β-catenin and Survivin were detected by real-time PCR and Western blot. Results The cisplatin-resistance cell line A549/DDP was constructed successfully, and its cisplatin-resistance is 13.7 times higher than in A549. There was no significance difference between A549 and A549/DDP in cell proliferation. The expression level of SOX4 is higher in A549/DDP than in A549. The cisplatin-resistance significantly decreased in A549/DDP cells after knockdown of SOX4 by siRNA transfection. The expres-sion level of β-catenin and Survivin significantly decreased in A549/DDP cells after knockdown of SOX4. Conclusion SOX4 can strengthen cisplatin-resistance of non-small cell lung cancer cell A549.

Background and objective It has been proven that chlorogenic acids can produce anticancer effects by regulating cell cycle, inducing apoptosis, inhibiting cell growth, Notch signaling pathways are closely related to many human tumors. The aim of this study is to study the mechanism of chlorogenic acid on apoptosis of non-small lung cancer through Notch1 pathway in animal level, and hope to provide theory basis on clinical treatment and research aimed at targeting Notch1 signaling in non-small cell carcinoma (NSCLC).Methods MTT assay was used to evaluate the A549 cell proliferation under the treatment of chlorogenic acid. The effect of chlorogenic acid on apop-totic and cell cycle were detected by flow cytometry. The animal model of A549 cell transplanted in nude was estab-lished, tumer size and weight were detected. The mRNA level of Notch1 signal pathway related facter were detected by RT-PCR; the expression of Notch1 signal pathway related facter in tumor tissue was detected by western blot.Results Chlorogenic acid inhibited the A549 cell proliferation. incresed cell apoptotic and cell percentagein G2/M (P<0.05), and in a dose-dependent manner. In animal model, tumer size and weight were lower than control group, the difference was statistically significant (P<0.05). The relative expression of mRNA of Notch1, VEGF, Delta4, HES1 and HEY1 were decreaced (P<0.05) in tumor tissue which treated with chlorogenic. The expression of Notch1 were decreaced, PTEN, p-PTEN, p-AKT were increced significantly in tumor tissue which treated with chlorogenic (P<0.05).Con-clusion Chlorogenic acid can regulate theapoptosis of non-small lung cancer through Notch pathway in animal level,which may be associated with the down-regulating the expression of VEGF and Delta4. Notch pathway may cross talk with PI3K/AKT pathway through PTEN in NSCLC.

Objective:To evaluate the in vitro cross-neutralization of serum antibodies in human and mice immunized with inactivated SARS-CoV-2 vaccine against Delta and Beta variants. Methods:Human serum samples after a second and a third dose of inactivated SARS-CoV-2 vaccine and mouse serum samples after a two-dose vaccination were collected. The neutralizing antibodies in the samples against SARS-CoV-2 strains of prototype, Delta and Beta variants were detected using micro-neutralization assay in biosafety level Ⅲ laboratory. The seroconversion rates and geometric mean titers (GMTs) of antibodies were calculated.Results:The seroconversion rates of antibodies in human serum samples against different SARS-CoV-2 strains were all above 95%. After two-dose vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 109, 41 and 15, respectively. The GMTs decreased by 2.7 folds and 7.3 folds for the Delta and Beta variants as compared with the prototype strain. After the booster vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 446, 190 and 86, respectively. The GMTs of neutralizing antibodies against Delta and Beta variants decreased by 2.3 folds and 5.2 folds as compared with that against the prototype strain. The seroconversion rates of antibodies against different SARS-CoV-2 strains in mouse serum samples were all 100%. The GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 2 037, 862 and 408, respectively. The GMTs decreased by 2.4 folds and 5.0 folds for the Delta and Beta variants.Conclusions:Inactivated SARS-CoV-2 vaccine could induce a certain level of neutralizing antibodies against Delta and Beta variants in both human and mouse models. Moreover, a third dose of vaccine induced higher levels of neutralizing antibodies against Delta and Beta variants in human. This study provided valuable data for the clinical application and protective evaluation of the inactivated SARS-CoV-2 vaccine.

OBJECTIVE: To explore the relationship between extend types and distant metastasis of nasopharyngeal carcinoma (NPC). METHOD: Retrospective analyze 260 patients with nasopharyngeal carcinoma, among which 162 cases were distant metastasis (metastasis group) and 98 cases were neither distant metastasis nor recurrence (disease-free group) over 5 years after radiotherapy. All these patients were staged depending on CT or MRI image before treatment and divided into local-regional type(T(1-2)N(0-1)) for 36 cases and upward invasion type (T(3-4)N(0-1)) for 68 cases and downward invasion type (T(1-2)N(2-3)) for 75 cases and mixed type (T(3-4)N(2-3)) for 81 cases. The differences between two groups was analyzed using Chi-square test. RESULT: The local-regional type and upward invasion type was 25.3% for the metastasis group and 64.3% for the disease-free group. The downward and mixed invasion was 74.7% for the metastasis group and 35.7% for the disease-free group. The rate(proportion) of N(2-3) was significantly higher in metastasis group than in disease-free group with limited extension (84.4% vs. 33.3%, P < 0.01). The rates(proportion) of N(0-1) and N(2-3) were also significantly higher in metastasis group than in disease-free group with severity extension (T(3-4) (60% vs. 36.1% and 68.4% vs. 40%, P < 0.01). CONCLUSION The extent of cervical lymph node metastases is one of the most important factors of NPC with distant metastasis, severity extension of primary disease should also be in consideration. Even the limitations of primary disease, once cervical lymph node metastasis occurs, the risk of distant metastasis is significantly increased.
Adolescent ; Adult ; Aged ; Carcinoma ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; Neck ; Neoplasm Metastasis ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Young Adult