OBJECTIVE: To investigate the relationship of the expression levels of protein arginine methyltransferase 5 (PRMT5) and Dickkopf-related protein 3 (DKK3) in the PCa tissue with biochemical recurrence (BR) of the malignancy after radical surgery. METHODS: This study included 105 cases of PCa diagnosed in our hospital from January 2016 to December 2020 and, according to BR within 3 years after surgery, we divided them into a BR (n = 22) and a non-BR group (n = 83). We detected the expressions of PRMT5 and DKK3 in the prostate tissues of the patients by immunohistochemistry, analyzed the correlation of the expression levels of PRMT5 and DKK3 using the Spearman method, and conducted a multivariate analysis of postoperative BR of the malignancy using the Cox multivariate regression model. RESULTS: The positive expression of PRMT5 was significantly higher while that of DKK3 remarkably lower in the PCa than in the adjacent tissue (P<0.05). Spearman correlation analysis showed a negative correlation between the expression levels of PRMT5 and DKK3 in the PCa tissue (r = －0.532, P<0.05). The expressions of PRMT5 and DKK3 were significantly associated with tumor-node-metastasis (TNM) stages, positive surgical margins, peripheral nerve invasion, capsular invasion, seminal vesicle invasion and vascular invasion (P<0.05). The percentage of TNM stages III－IV, the positive expression of PRMT5 and the negative expression of DKK3 were remarkably higher in the BR than in the non-BR group (P<0.05). PRMT5 was found to be an independent risk factor for while DKK3 a protective factor against postoperative BR of PCa in the patients (P<0.05). CONCLUSION PRMT5 is highly while DKK3 lowly expressed in PCa tissue, and their expressions are both closely related to the biochemical recurrence of PCa after radical surgery.
Humans ; Male ; Protein-Arginine N-Methyltransferases/metabolism* ; Prostatectomy ; Prostatic Neoplasms/pathology* ; Neoplasm Recurrence, Local ; Adaptor Proteins, Signal Transducing ; Intercellular Signaling Peptides and Proteins/metabolism* ; Middle Aged ; Immunohistochemistry

Objective: To evaluate the safety and clinical efficiency of holmium laser enucleation of the prostate (HoLEP) in the treatment of small-volume BPH (SBPH) complicated by severe lower urinary tract symptoms (LUTS). METHODS: We retrospectively analyzed the clinical data on 82 cases of SBPH with severe LUTS treated by HoLEP from January 2017 to December 2018. The patients were aged (65.5 ± 7.6) years, with a mean prostate volume of <40 ml, a total IPSS of 24.8 ± 4.6, a QOL score of 5.2 ± 0.8, the maximum urinary flow rate (Qmax) of (7.6 ± 3.7) ml/s, and a mean PSA level of (1.8 ± 1.4) μg/L. RESULTS: All the operations were successfully completed, the mean operation time averaging (30.2 ± 5.0) min, enucleation time (26.7 ± 5.6) min and comminution time (3.5 ± 1.1) min, and the enucleated tissue weighing (20.3 ± 4.9) g. After surgery, the bladders were irrigated for (3.5 ± 1.9) h, with (3.0 ± 1.7) L of rinse solution, and catheterization lasted (24.8 ± 9.7) h. Histopathology revealed moderate or severe lymphocytic infiltration in 69 cases (84.1%). At 6 months after operation, significant improvement was observed in the IPSS, QOL, Qmax and PSA level compared with the baseline (P < 0.05). To date, no urethral stricture-related reoperation was ever necessitated. CONCLUSIONS HoLEP is safe and effective for the treatment of SBPH complicated by severe LUTS and can be employed after adequate preoperative evaluation of the patient.《.
Humans ; Lasers, Solid-State ; Lower Urinary Tract Symptoms/surgery* ; Male ; Prostate/surgery* ; Prostatic Hyperplasia/surgery* ; Quality of Life ; Retrospective Studies

Astragali Radix is commonly used as bulk medicinal materials. Chinese Pharmacopoeia contains about 150 compound preparations of Astragali Radix, but the sample preparation method under the determination of Astragali Radix content in Chinese Pharmacopoeia is tedious and time-consuming, not convenient for the test of a large number of samples. Therefore, it is of great significance to simplify the sample preparation method and improve the practicability of the method for the quality control of Astragali Radix and its preparations. In this study, ultrasonic extraction method was used instead of heated reflux extraction, and solid phase extraction method was used to enrich and prepare the samples. A set of practical quality evaluation method was established for Astragali Radix slices and standard decoction, greatly shortening the sample preparation time and improving the accuracy of the method. The results of Astragali Radix standard decoction analysis showed that the transfer rate of calycosin 7-O-β-D-glucospyranoside,(96.5±28.7)%, had great variation, which was found to be related to the conversion of mulberry isoflavone glucoside into calycosin 7-O-β-D-glucospyranoside during the preparation of standard decoction. The transfer rates were(59.4±14.4)% and(101.3±12.3)% for calycosin and astragaloside Ⅳ respectively, which were relatively stable. Therefore, it is suggested that Astragali Radix slices and water decoction preparations should be evaluated by using calycosin and astragaloside Ⅳ as the quality evaluation index. The results provide a scientific and practical method for quality control of Astragali Radix slices and its standard decoction, and also provide scientific evidence for quality evaluation of the preparations.
Astragalus Plant/chemistry* ; Astragalus propinquus ; Drugs, Chinese Herbal/standards* ; Glucosides/analysis* ; Plant Roots/chemistry* ; Quality Control ; Solid Phase Extraction

Objective:To investigate the influence of Portulaca extracts on H1R,PAR-2 and TRPV1 in skins of rats and the regulation mechanism of H1R,PAR-2/TRPV1 itch signal pathway with atopic eczema.Methods:30 SD rats were randomly divided into normal control group,model group and Porulaca group,each group contained 10 rats.In addition to the normal group,2,4-dinitrochloro-benzene was used on the rest of group for making rat model with atopic eczema.After the success of the model manufacture,each group was given corresponding drugs.After the last administration,EASI was evaluated.The levels of H1R,PAR-2 and TRPV1 in rats skins were detected by immunohistochemical method.Ca2+concentration was analyzed by flow cytometry.Results: Compared with normal control group,EASI(P<0.01),the levels of H1R(P<0.05) and PAR-2(P<0.01),and Ca2+concentration(P<0.01) of rats in model group were significantly increased.The levels of TRPV1 were not increased obviously (P>0.05).Compared with model group,EASI (P<0.01),the levels of H1R(P<0.01) and PAR-2(P<0.01),and Ca2+concentration(P<0.05) of rats in Porulaca group were sig-nificantly reduced,and the levels of TRPV1 were not decreased obviously(P>0.05).Conclusion:Purslane could reduce the levels of H1R,PAR-2 and Ca2+concentration to treat the acute eczema.The mechanism may be to lose the activation of downstream molecule TRPV1 and reduce the inflow of Ca2+by reducing the levels of upstream molecules H1R and PAR-2,then to achieve the effect of anti itch.

Background:The Food and Drug Administration recently announced that the use of morcellation may cause fibroids or pelvic dissemination and metastasis of uterine sarcoma;therefore,the use of morcellation is limited in the USA.A large sample study is necessary to assess the proportion of uterine malignant tumors found in patients with laparoscopic myomectomy.Methods:A national multicenter study was performed in China.From 2002 to 2014,33,723 cases were retrospectively selected.We calculated the prevalence and recorded the clinical characteristics of the patients with malignancy after morcellation application.A total of 62 cases were finally pathologically confirmed as malignant postoperatively.Additionally,the medical records of the 62 patients were analyzed in details.Results:The proportion of postoperative malignancy after morcellation application was 0.18％ (62/33,723) for patients who underwent laparoscopic myomectomy.Nearly 62.9％ (39/62) of patients had demonstrated blood flow signals in the uterine fibroids before surgery.And,23 (37.1％) patients showed rapid growth at the final preoperative ultrasound.With respect to the pathological types,38 (61.3％) patients had detectable endometrial stromal sarcoma,13 (21.0％) had detectable uterine leiomyosarcoma,only 3 (3.2％) had detectable carcinosarcoma,and 5 (8.1％) patients with leiomyoma had an undetermined malignant potential.Conclusions:The proportion of malignancy is low after using morcellation in patients who undergo laparoscopic myomectomy.Patients with fast-growing uterine fibroids and abnormal ultrasonic tumor blood flow should be considered for malignant potential,and morcellation should be avoided.

OBJECTIVES: To recognize the possibility of Y fragment deletion of Amelogenin gene intuitively and simply according to the genotyping graphs. METHODS: By calculating the ratio of total peak height of genotyping graphs, the statistics of equilibrium distribution between Amelogenin and D3S1358 loci, Amelogenin X-gene and Amelogenin Y-gene, and different alleles of D3S1358 loci from 1 968 individuals was analyzed after amplified by PowerPlex^® 21 detection kit. RESULTS: Sum of peak height of Amelogenin X allele was not less than 60% that of D3S1358 loci alleles in 90.8% female samples, and sum of peak height of Amelogenin X allele was not higher than 70% that of D3S1358 loci alleles in 94.9% male samples. CONCLUSIONS The result of genotyping after amplified by PowerPlex^® 21 detection kit shows that the possibility of Y fragment deletion should be considered when only Amelogenin X-gene of Amelogenin is detected and the peak height of Amelogenin X-gene is not higher than 70% of the total peak height of D3S1358 loci.
Alleles ; Amelogenin/genetics* ; Asian People/genetics* ; DNA Fingerprinting/methods* ; Female ; Genotype ; Humans ; Male ; Mutation ; Polymerase Chain Reaction/methods* ; Population Groups

Tilianin was separated and authenticated from the seeds of Dracocephalum moldavia, a Uygur medicine, by chromatographic technique and spectroscopic method. The purity of tilianin is more than 98% determined by HPLC area normalization method. Thin layer chromatography (TLC) method was used to separate tilianin from D. moldavia by mixture of chloroform-methanol (5: 1) as a developing solvent on high performance silicagel precoated plate (SGF254) and using aluminium trichloride as a chromogenic agent for qualitative identification of D. moldavia. To establish a HPLC method for quantitative analysis of D. moldavia, tilianin was used as a Quantitative marker and separated on a C18 (4.6 mm x 250 mm, 5 μm) column with acetonitrile-01% formic acid (25: 75) as the mobile phase and detected at 330 nm. The calibration curve of tilianin displayed ideal linearity over the range of 0.617 2-123.44 μg x mL(-1) with a regression equation of Y = 33.773X - 0.824 8 (r = 1). The average recovery of tilianin was 101.0% with RSD of 3.7%. The RSD values of intra-day and inter-day precision were less than 2%. The content of tilianin in 4 batches of the authenticated semen of D. Moldavia was between 0.016 and 0.187 mg x g(-1). The qualitative and quantitative method established is suitable for the quality evaluation and assessment of semen of D. Moldavia.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Flavonoids ; chemistry ; isolation & purification ; standards ; Glycosides ; chemistry ; isolation & purification ; standards ; Lamiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Quality Control

OBJECTIVETo investigate the effets of flurothyl-induced neonatal recurrent seizures on glucocorticoid receptor (GR) expression in the rat brain.

METHODSForty-eight seven-day-old Sprague-Dawley rats were randomly divided into two groups: control and seizure. Seizures were induced by inhalant flurothyl daily for six consecutive days. Brains were sampled on postnatal days 13, 15 and 19. The expression of GR protein in the cerebral cortex was detected by Western blot and immunohistochemical method.

RESULTSThe expression of GR in the cerebral cortical plasma protein was significantly lower in the seizure group than in the control group on postnatal day 15. The expression of GR protein in the cerebral cortical nuclear protein decreased significantly in the seizure group compared with that in the control group on postnatal days 15 and 19 (p<0.05). Compared to the control group, the accumulated optical density (AOD) of GR immunoreactivity (IR) decreased significantly in the parietal cortex on postnatal day 13 (p<0.05), the AOD of GR IR decreased significantly in the parietal cortex and the temporal cortex on postnatal day 15 (p<0.05), and the AOD of GR IR decreased significantly in the parietal cortex, temporal cortex and the frontal cortex in the seizure group on postnatal day 19 (p<0.05).

CONCLUSIONSRecurrent seizures in neonatal rats result in abnormal GR expression in the cerebral cortex which might play an important role in short-term brain injury induced by early recurrent seizures.

Animals ; Blotting, Western ; Cerebral Cortex ; chemistry ; Female ; Hypothalamo-Hypophyseal System ; physiology ; Immunohistochemistry ; Male ; Pituitary-Adrenal System ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; analysis ; physiology ; Recurrence ; Seizures ; metabolism

Objective To discuss whether the separating block of sensory nerve with acupoint-injection and epidural analgesia by ropivacaine during labour.Methods Ninty full term primigravidae with ASAⅠ～Ⅱwere randomly divided into 3 groups:group E(n = 30),group C(n = 30)and group A(n = 30).When the external cervi- cal os was dilated to 3cm,group A:acupiont-injection to Ci Liao;group E:epidural analgesia with 0.075% ropiva- caine and fentanyel 2?g/ml;group C:combining group A and group E except fentanyel 2?g/ml.Level of pain(VAS scores),degree of sensory and motor block,the mode of delivery,the progress of labour and side effects were recorded and compared.Results The level of pain(VAS scores)in group A was higher than that of group C and group E,and there were no significant differences in group C and group E,Bur there were some side effects discovered in group E. There was no significant differences of the mode of delivery in the groups.Conclusion The method that acupoint- rejection combined with epidural analgesia by 0.075% ropivacaine is effective and has no side effects for labour.

Apolipoprotein AI (apo AI), the major protein component of human high-density lipoprotein (HDL), is a single-chain polypeptide of 243 amino acids. Several epidemiological studies have shown that the plasma concentrations of HDL has the role of reverse cholesterol transport (RCT) and inversely correlated with the incidence of coronary artery disease. Because apo AI lacks post-translational modifications, it is convenient to express human apo AI in Escherichia coli expression system. However, there is a poor stability of the mRNA and the apo AI protein in E. coli, it is difficult to express mature apo AI in recombinant bacteria, moreover, even as a fusion protein, apo AI is still sensitive to degradation and can not be cleaved efficiently from the fusion tags. In contrast, proapolipoprotein AI (proapo AI, having an additional polypeptide containing the amino acids Arg-His-Phe-Trp-Gln-Gln at the amino-teminal of the mature protein) proved stable and undegraded in Escherichia coli, and therefore, in this research, an expression system of E. coli including a plasmid of P(R)P(L) tandem promoter was adapted to produce proapo AI. Furthermore, site-directed mutagenesis of the proapo AI cDNA was performed to generate a Clu8Asp mutation in the amino-terminal sequence of proapo AI which created an acid labile Asp-Pro peptide bond between amino acid 8 and 9, and permitted specific chemical cleavage to remove pro-peptide. After inducing with a shift of temperature, yields of recombinant proapo AI achieved about 40% of total cell protein and the recombinant proapo AI expressed proved as a form of inclusion body in cells, so protein need to renature. First of all, the protein was dissolved in buffer with denaturant, and renaturation was carried out on a hydrophobic interaction column (Phenyl Sepharose), ion-exchange chromatography and gel-filtration chromatography were then used to further purify the protein. The purified recombinant apo AI was detected by a set of tests including Western-blotting, Circular dichroism spectra and lipid-binding test, the results shown that recombinant apo AI has similar structural and lipid-binding properties identical to those of native plasma apo AI, which facilitates further research and application.
Apolipoprotein A-I ; biosynthesis ; genetics ; Chromatography, Ion Exchange ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Protein Precursors ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification