OBJECTIVETo investigate the changes on the number and distribution of myoepithelial cells (MECs) during parotid gland regeneration.

METHODSA total of 54 Wistar rats were divided into eight experimental groups and one normal control group, with six rats in each group. The right parotid ducts of the rats in the experimental groups were ligated for 14 days and then reopened. The parotid tissue specimens were harvested at days 0, 1, 3, 5, 7, 10, 14, and 21. The histological changes of the regenerating gland were examined using hematoxylin-eosin staining. Immunohistochemical labeling was also performed to investigate the changes in the number and distribution of MECs at different time points of parotid regeneration. Both the histological and immunohistochemical changes observed in the experimental groups were compared with those in the normal control group.

RESULTSThe parotid gland showed marked atrophy 14 days after ligation. Most acinar cells disap- peared, but the number of duct-like structures obviously increased. MECs apparently increased in number and were mainly located at the periphery of the duct-like structures. Three days after duct reopening, the number of acinar cells significantly increased and the duct-like structures significantly reduced. Meanwhile, MECs also decreased in number and were mainly located at the periphery of the newly formed acini and duct-like structures. The number of MECs noticeably decreased 3 and 5 days after duct reopening. At 14 days after duct reopening, the glandular structures and the number and distribution of MECs returned to normal compared with those in the normal control group.

CONCLUSIONThe number and distribution of MECs returned to normal condition after parotid gland atrophy. Parotid regeneration mainly occurred within 5 days after duct reopening.

Objective:To investigate the histological changes of parotid gland after ligation and reopening of the main duct.Meth-ods:The right parotid main duct was ligated for 14 days and then reopened,the specimen were harvested at day 0,1,3,5,7,10,14 and 21 after reopening(n=5 )respectively.By HE staining and Morphology measurement,the morphological and volume fraction changes of acinus,duct and mesenchyma were observed and measured.Results:The parotid gland atrophied significantly 14 days af-ter the duct ligation.From day 3 after duct reopening,the acinar volume fraction continued to increase,both the duct and the mesen-chymal tissue volume fraction continued to decrease significantly(P<0.05).After 14 days of duct reopening,there was no signifi-cant difference in volume fraction of acinar,duct and mesenchyma,compared with those of normal control group(P>0.05).Conclu-sion:After reopening of the ligated main duct the atrophied parotid gland is able to regenerate to be normal.

Objective To investigate the optimal process conditions for the separation and purification of extract from Hyssopus cuspidatus Boriss. by polyamide resins. Methods The total flavonoids and rosmarinic acid were used as the indexes. The maximum amount of sample solution, elution volume, concentration of sample solution, adsorption time of resin, loading time of sample solution and the amount of eluting solvent, pH and elution rate in the resin purification process were screened by single factor method. Results The optimal purification parameters were as follows: 10 mg/mL of extract, 12 mL of sample amount, 2 BV of water to remove impurities, 40% ethanol to elute 9 BV; the concentration of rosmarinic acid in sample solution was 86.3 μg/mL, and the total flavonoid concentration was 117.8 μg/mL; the resin adsorption time was 14 h; the pH of sample solution was 6.5; the elution rate was 3.0 BV/h. Conclusion This method is simple and feasible, fit for separating and purifying of extract from Hyssopus cuspidatus Boriss.

OBJECTIVE:To provide reference for clarifying the pharmacodynamic material basis of anti-inflammation effect of Uygur medicine Hyssopus cuspidatus. METHODS:HPLC was conducted to establish the fingerprint of 8 extracts with different po-larities. Using macrophage RAW264.7 as object,the inhibitory effect of extracts with different polarities on inflammatory factor ni-tric oxide(NO),TNF-α and IL-6 in supernatant of cell culture medium was detected,the differences of anti-inflammatory activity in vitro were compared. Grey correlation analysis method was used to analyze the spectrum-effect relationship of peak area of common peaks and anti-inflammation activity. RESULTS:The fingerprint of 8 extracts with different polarities showed obvious differences. Anti-inflammation in vitro results suggested that 30% ethanol extract had the strongest inhibitory effect on inflammatory cytokines in vitro. There were 14 common peaks in the established HPLC fingerprints,the 5 common peaks that was closely related to the in-hibitory effect of inflammatory factors in vitro were peak 8,6,5,10,13,respectively;peak 8 was rosmarinic acid. CONCLUSIONS:The established fingerprint and its spectrum-effect relationship with anti-inflammation activity in vitro can provide certain reference for its pharmacodynanic material basis study.

OBJECTIVE:To investigate the effects of different elutions of Hyssopus cuspidatus on smooth muscle contraction of isolated tracheal in Guinea pigs. METHODS:Isolated tracheal rings were prepared and soaked in Krebs-Henseleit,using acetyl-choline(ACh,1×10-7 g/mL)or histamine(His,1×10-6 g/mL)to induce contraction of tracheal rings,then the effects of H. cuspi-datus water elution and 30%,50%,60%,70%,95%ethanol elutions with mass concentrations of 0.08,0.16,0.32,0.64,1.28, 2.56 mg/mL on contraction of tracheal rings were respectively investigated. Contraction curves were recorded and antispasmodic rates were calculated. Tests were treated with saline as blank control and aminophylline (0.08 mg/mL) as positive control. RE-SULTS:Compared with blank control,0.16-2.56 mg/mL 30%,50% ethanol elution,0.32-2.56 mg/mL 60% ethanol elution and 0.64-2.56 mg/mL 70%,95% ethanol elution can obviously inhibit ACh-induced contraction of tracheal rings,antispasmodic rates were obviously increased(P<0.05 or P<0.01);0.32-2.56 mg/mL water elution and 30%,50% ethanol elution,0.16-2.56 mg/mL 60%,70% ethanol elution and 1.28-2.56 mg/mL 95% ethanol elution can obviously inhibit His-induced contraction of tracheal rings,antispasmodic rates were obviously increased (P<0.05 or P<0.01). The effects of 2.56 mg/mL 60% ethanol elution ap-proach to aminophylline. CONCLUSIONS:Different elutions of H. cuspidatus has certain antagonistic effect on the ACh-induced or His-induced smooth muscle contraction of isolated tracheal in Guinea pigs;60% ethanol elution shows the strongest effect, which has similar effects with aminophylline at high mass concentration.

AIM:To analyze circulating miR-141 in the serum as a non-invasive biomarker in the patients with prostate cancer ( PCa) and benign prostate hyperplasia ( BPH) , and healthy individuals.METHODS: A total of 75 pa-tients with PCa, 52 with BPH and 40 healthy individuals were enrolled into this study.Total RNA was isolated from the se-rum samples and the circulating levels of miR-141 were determined using quantitative real-time polymerase chain reaction. RESULTS:The serum levels of miR-141 were significantly higher in the patients with PCa compared to the patients with BPH and the healthy controls (P<0.01).The level of miR-141 in PCa group obviously differed from that in BPH group and healthy control group with high diagnosis performance, with areas under the curve of 0.785 and 0.801, respectively. No statistically significant difference of the serum miR-141 levels between the patients with BPH and healthy individuals was observed (P>0.05).The serum miR-141 level was also found to be related to Gleason score, clinical stage and bone me-tastasis status of the patients with PCa (P<0.05), and the patients with higher Gleason scores had higher serum miR-141 levels.No relationship was detected between miRNA-141 level and the patient’ s age, biochemistry recurrence and serum prostate-specific antigen level (P>0.05 for all comparisons).CONCLUSION: Circulating miR-141 could serve as a non-invasive biomarker for prostate cancer diagnosis, staging and prognosis prediction.

Objective To explore the clinical effect of mycobacterium tuberculosis determination by culture and microscopy assay from sputum specimens of pulmonary tuberculosis patients.Methods The sputum specimens were dyed with the Ziehl-Neelsen anti-acid dyeing direct microscopy and examined by BACTEC-TB 960 culture system.Results The positive rate of culture and microscopy were 36.6% and 24.6%,the difference in the positive rate between the two methods was significant(P

OBJECTIVE:To compare the differences between artificially cultivated and wild Xinjiang Artemisia rupestris,and screen the different components. METHODS:HPLC-MS was adopted to establish the fingerprints of artificially cultivated and wild Xinjiang A. rupestris from different origin and harvest time. Principal component analysis was conducted by Marker ViewTM soft-ware and SIMCA-P 11.5 software,the characteristics of principal components were analyzed,difference variable was screened, and different components of artificially cultivated and wild varieties were obtained. RESULTS:Fingerprints of 22 batches of A. rup-estris(12 batches of wild varieties,10 batches of artificially cultivated varieties)were established. According to the principal com-ponent analysis,artificially cultivated and wild varieties were well grouped,with obvious differences;the principal components of artificially cultivated varieties with different harvest time showed certain difference,mainly before and after flowering,concentrat-ing in to-be flowering and full flowering periods. Wild varieties from different origins had obvious regional difference,showing cer-tain differences in composition and content. 268 variables were found in matrix of positive ion mode and 155 in negative ion mode. 28 groups of variables were extracted by difference variable,and 19 variables were determined. CONCLUSIONS:Artificially culti-vated and wild varieties have obvious difference in principal component,mainly in flowering period and picking places. It can pro-vide theoretical basis for the standardized cultivation and origin protection of Xinjiang A. rupestris.

Objective:To investigate the expression and correlation of Bcl-2 and Bax in the parotid gland after leading duct ligation in rats. Methods:Atrophy of the right parotid was induced by ligating the right main duct of 72 rats. Immunohistochemical labelling was performed to study the expression of Bcl-2 and Bax 1, 3, 5, 7, 14, 21, 30, 60, 90, 150 and 180 days after duct ligation. Results:7 d after duct ligation most acinar cells disappeared. The distribution of Bcl-2 and Bax protein in normal parotid was in cytoplasm with unifo-maity. Bcl-2 and bax higher expression was identified in the gland at all time points. The expression of Bcl-2 protein was significantly in-creased and reached the peak at 21 d after duct ligation. More Bax-positive acinar cells on day 3 were observed, then the expression of Bcl-2 and Bax protein was descended. Higher Bcl-2/Bax ratio was identified at 1-21 d, then descended. Conclusion:The expression of Bcl-2 and Bax is associated with the atophy of the parotid glad after rat parotid duct ligation.

Objective To develop an assay of PCR-produet direct sequencing to detect hepatitis B virus (HBV) YMDD mutation, and compare the results gained by the sequencing and traditional real-time fluorescent PCR assays. Methods Serum samples were collected from 103 patients with chronic hepatitis B. HBV DNA were extracted from sers. YMDD mutation was detected by a commercial real-time PCR assay. Meanwhile, HBV reverse transcriptase-encoding gene was amplified by a nested PCR assay. The PCR products were directly subjected to sequencing at two directions, and the sequencing results were analyzed by NTI program. Using Kappa test, comparison was made between the results of rtM204-site mutations obtained by the direct sequencing and YMDD mutations by the real-time fluorescent PCR. Results The direct sequencing assay proved to be highly effective with bread range of detection in viral load from 500 to 1010copies/ml. And it may simultaneously avoid inhibitory effect caused by high viral load. The coincidence rates between two assays were 100% for YIDD, 97. 1% for YVDD, 76. 2% for YIDD/YVDD coexistence (Kappa = 0. 853, P < 0. 01). Conclusions The direct sequencing assay for HBV drug-resistant mutation detection is highly sensitive with broad dynamic range. It has high coincidence rate with real-time fluorescent PCR assay with advantage of detecting YMDD, YIDD and YVDD mutations simultaneously.