OBJECTIVE: To establish the rapid PCR amplification program and system and to verify the technical indexes. METHODS: PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as A melogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed. RESULTS: The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate. CONCLUSION The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.
Electrophoresis, Capillary ; Fluorescence ; Genetics, Population ; Genotype ; Humans ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction/methods* ; Sensitivity and Specificity

Objective To find out severity and types of syphilis infection in blood donors, attendants and persons to have their premrital medical examination in Zhoushan City and offer a new measure for prevention and treatment of syphilis. Methods Totally, 174 589 blood donors, attendants and persons to have their premarital medical examinations were screened preliminarily for inapparent syphilis with non-TPHA, and then TPHA was applied to confirm the diagnosis, according to the National Standard No. GB 15974-1995, combining with clinical symptoms and physical check-up. Results A total of 1 327 cases of syphilis from 174 589 samples tested, including blood donors, attendants and persons to have their premarital medical examinations, were diagnosed, with an inapparent infection rate of 7. 60‰ in average, 6. 42‰in males and 8. 74‰ in females, with a sex ratio of 0.71 (X2 = 29. 92, P

OBJECTIVETo observe the effect of CKJ Recipe (consisting of Cordyceps sinensis polysaccharide, amygdaloside, and gypenosides) containing serum on the activation of rat primary hepatic stellate cells (rHSCs) and to explore its pharmacological mechanism.

METHODSrHSCs were isolated form liver and cultured for four days. Then they were divided into the normal control group, the model group, and the CKJ group. rHSCs in the model group and the CKJ group were treated with 2.5 ng/mL transforming growth factor beta1 (TGF-beta1) in serum-free DMEM for 24 h. Serum free DMEM (containing no TGF-beta1) was taken as the control for the normal control group. rHSCs in the CKJ group were treated with 5% CKJ-containing serum for 24 h. rHSCs in the other two groups were treated with 5% blank serum for 24 h.The protein expression level of a smooth muscle actin (alpha-SMA) was determined using high throughput screening (HCS) and Western blot. mRNA expression levels of alpha-SMA, collagen I (Col-I), platelet-derived growth factor receptor beta (PDGF-betaR), TGF-beta1, transforming growth factor beta receptor 1 (TGF-betaR1), and transforming growth factor beta receptor 2 (TGF-beta R2) were detected using quantitative RT-PCR.

RESULTSCompared with the normal control group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-betaR1, and TGF-betaR2 significantly increased in the model group (P<0.05, P<0.01). Compared with the model group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-beta1, and TGF-beta R2 significantly decreased in the CKJ group (P<0.05, P<0.01).

CONCLUSIONCKJ containing serum could inhibit the protein expression level of o-SMA, which was probably related with inhibiting TGF-beta1 and its related receptors.

Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hepatic Stellate Cells ; metabolism ; Rats ; Transforming Growth Factor beta ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo undergo tinnitus evaluation test and masking therapy of subject tinnitus associated with sensorineural hearing loss as a reference for diagnosis and guiding masking therapy.

METHODSThe 66 patients with subject tinnitus were diagnosed as sensorineural hearing loss. Sixty-six patients divide into three groups according to the results of pure tone audiometry, including steep drop type in 28 patients, slow drop type in 20 patients, and flat type in 18 patients. All the patients underwent tinnitus evaluation tests (pitch matching, intensity matching, tinnitus masking curves, residual inhibition) and masking therapy.

RESULTSTinnitus with steep drop type manifest was as low intensity [average intensity (5.7 +/- 2.9) dB (x(-) +/- s)] and high frequency (median with 4750 Hz). Residual inhibition was almost positive, but was usually consistent with convergence and congruence tinnitus masking curves. Masking therapy had better effect in the treatment of this type of tinnitus (effective rate 89.3%). Tinnitus with slow drop type manifest was as low intensity [average intensity: (6.2 +/- 4.8) dB] and high frequency (median: 4050 Hz). The distribution of residual inhibition and tinnitus masking curves had no obviously characteristics. The effective rate of masking therapy of this type of tinnitus (55.0%) was higher than tinnitus with flat type but low than that of tinnitus with steep drop type. The intensity of tinnitus with flat type [average intensity: (9.2 +/- 5.0) dB] was higher than that of the previous groups. The distribution of frequency of this type had no obviously characteristics. The residual inhibition was almost negative, and was usually consistent with divergence and persistence tinnitus masking curves. Masking therapy had unsatisfactory curative effect in the treatment of this type of tinnitus (effective rate 11.1%). The effective rate of masking therapy was significant differences among the three groups (chi(2) = 9.127, P < 0.05).

CONCLUSIONSFor the tinnitus patients with steep drop type audiometric curve, they are recommended masking therapy. For the tinnitus patients with slow drop type audiometric curve, masking therapy may be attempted to perform. For the tinnitus patients with flat type audiometric curve, they do not recommend the masking therapy.

Adolescent ; Adult ; Aged ; Audiometry, Pure-Tone ; Female ; Hearing Loss, Sensorineural ; diagnosis ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Perceptual Masking ; Tinnitus ; diagnosis ; physiopathology ; therapy ; Young Adult

Thirty SHRs were obtained randomly to hypertension, model group, captopril group and Qingre jiangya capsule group. Ten Wistar rats were used as control group. The hippocampus tissue was removed to explore the damage of spontaneously hypertensive rats (SHR) and the protective effect of Qingre jiangya capsule after continuously administered for 14 days. And then the data were processed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). The research results revealed captopril group was significantly different from the other three groups. The classification of other three groups is also very clear after captopril group removed. This suggested that Qingre jiangya capsule could improve the overall metabolism compared with captopril. Four metabolites were identified: dimethylglycine, glycerophosphocholine, aldosterone and noradrenaline. Hypertension hippocampus damage may mainly be expressed in tyrosine metabolism, aldosterone-regulated sodium, vascular smooth muscle contraction reabsorption, and Qingre jiangya capsule could reverse the hippocampus tissue damage of SHR.
Animals ; Capsules ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; drug effects ; Hypertension ; drug therapy ; Male ; Rats ; Rats, Inbred SHR ; Rats, Wistar

Objective To observe toxic effect of deoxynivalenol(DON)on articular cartilage and synovium of New Zealand rabbits's knee ioints.Methods Fifteen male rabbits were divided randomly into 3 groups:control, high-dosage,and low-dosage group.In high-dosage and low-dosage group,saline solution of DON was injected with a dose of 0.10 and 0.05 ms/kg every 48 h into ear vein of rabbits.Specimen of articular cartilage and synovium were through pathologY methods,and IL-1β,TNF-α,NO levels were assayed in joint liquid,after 20 days. Results Morphological changes were observed, such as synovium inflammative infiltration, chondrocytes deformation and necrosis under light microscope.The levels of IL-1β,TNF-α and NO had statistical significance in comDarison between 3 grouPs(F=19.396,18.195,22.136,P＜0.05).The levels of IL-1β,TNF-α and NO were significantly higher(all P＜0.05),high-dosage[(0.451±0.091),(0.575±0.122)μg/L;(70.27±11.53)μmol/L] and low-dosage group[(0.295±0.107),(0.387±0.131)μg/L;(45.32±12.24)μmol/L]compared with control ((0.1 13±0.049),(0.138±0.087)μg/L;(23.56±9.35)μmoL/L],and high-dosage compared with low-dosage group Conclusions DON results in articular and synovial impairment,which has the symptom similar to osteoarthritis. DON probably causes osteoarthritis.

We predicted the anti-hepatitis B virus (HBV) active components and mechanism of Salvia miltiorrhiza based on network pharmacology. The active components of S. miltiorrhiza were obtained through TCMSP, PubChem database and literature research. The potential targets of the active components and HBV infection were predicted by SwissTargetPrediction and GeneCards databases, respectively. The protein-protein interaction (PPI) network was constructed by String database. Cytoscape software was adopted to construct a visual network of active component-disease target and perform topological analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using DAVID platform. The molecular docking of key components and core targets was carried out by AutoDock Vina software. We screened out a total of 38 active components and 178 disease-component overlapping targets. Enrichment analyses obtained 405 related GO items and 68 signaling pathways, such as T/B cell receptor signaling pathways, PI3K/AKT signaling pathway, and mTOR signaling pathway. According to the results of molecular docking, most characteristic components of S. miltiorrhiza (miltionone Ⅱ, miltirone, protocatechuic acid, lithospermic acid, protocatechualdehyde) showed good affinity with the key targets (PIK3CA, APP, STAT3,AKT1 and mTOR). Furthermore, the anti-HBV activity of lithospermic acid, the representative active component of S. miltiorrhiza, and its regulation on PI3K/AKT and mTOR signaling pathways were investigated in an HBV replicating mouse model. Animal welfare and experimental procedures follow the regulations of the Animal Ethics and Welfare Committee of Hubei University. The results showed that lithospermic acid significantly inhibited HBV DNA replication, reduced serum HBsAg and HBeAg levels, and decreased the phosphorylation protein expression levels of AKT and mTOR in liver, indicating that lithospermic acid might exert the anti-HBV activity by regulating PI3K/AKT and mTOR signaling pathways.

To investigate the cardioprotective effect of formononetin (FMN) on no-reflow (NR) after myocardial ischemia-reperfusion and its molecular mechanism based on integrated pharmacology and experimental verification, firstly, human breast cancer MCF-7 cells and myocardial NR rats were used to confirm the estrogenic activity and the effect of alleviating NR of FMN, respectively. Male SD rats were divided into Sham, NR, FMN (20 mg·kg^-1) and sodium nitroprusside (SNP, 5.0 mg·kg^-1) groups, which were administered once a day for one week, the experiment was approved by the Ethics Committee of Tianjin University of Traditional Chinese Medicine (TCM-LAEC2019095). The pharmacological analysis and in vivo study of NR rats were integrated to reveal the mechanism of FMN improving NR. The results showed that FMN had estrogenic effect and reduced NR by improving cardiac structure and function, reducing NR, ischemic myocardial area and pathological injury of cardiomyocytes. Integrated pharmacology predicts that the mechanism of FMN improving NR is mainly related to phosphatidyinositol-3-kinase-protein kinase B (PI3K-Akt) signal pathway. Phytoestrogens play a role in cardiovascular protection mainly by activating G protein-coupled estrogen receptor (GPER). GPER is also an important regulator in the upstream of PI3K-Akt signaling pathway. This study found that FMN can significantly activate GPER, p-PI3K, p-Akt and phospho-endothelial nitric oxide synthase (p-eNOS). It has good binding ability with GPER and eNOS protein. In this study, through the integration of pharmacology and experimental evaluation, it is revealed that FMN activates PI3K/Akt/eNOS signal pathway by activating GPER, thus significantly improving NR.

This study was aimed to evaluate the therapeutic efficacy of bortezomib combined with autologous peripheral blood hematopoietic stem cell transplantation (autoPBSCT) for patients with multiple myeloma (MM). 5 patients underwent autologous hematopoietic stem cell transplantation. Bortezomib treatment was supplied for patients before autoPBSCT and in the conditioning of transplantation, it was also used in maintaining treatment. Patients with transplantation adopted bortezomib plus melphalan conditioning regimen. The number of infused MNC and number of CD34(+) cells were 4.06×10(8) (4.09×10(8) - 4.37×10(8))/kg and 3.98×10(6) (2.49×10(6) - 8.2×10(6))/kg respectively. The results showed that hematopoiesis was reconstituted in 5 patients, with a neutrophil cell count more than 0.5×10(9)/L at day 14 (13 - 25 days) after transplantation and platelet count more than 50×10(9)/L at day 28 (21 - 41 days) after transplantation. Transplantation-associated death was not observed. 5 patients were disease-free survival. In conclusion, treatment of bortezomib combined with autologous peripheral hematopoietic stem cell transplantation is an effective method for patients with multiple myeloma. Use of bortezomib after transplantation might still be favourable to MM patients, for survival prolongation and life quality improvement.
Adult ; Boronic Acids ; therapeutic use ; Bortezomib ; Combined Modality Therapy ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; therapy ; Peripheral Blood Stem Cell Transplantation ; Pyrazines ; therapeutic use ; Transplantation Conditioning ; methods

OBJECTIVETo elucidate the possible mechanism of oral carcinogenesis and to explore the value of clinical application of the detection of cytokeratin (CK) 19 for oral squamous cell carcinoma (OSCC) patients.

METHODSThe cancerous tissues, para-cancerous tissues and excised lymph nodes were collected from 20 operated patients with OSCC. The patients didn't receive radiotherapy and chemotherapy before hospitalization. The relative expression of CK19 mRNA in those tissues was detected by fluorescent quantitative polymerase chain reaction (FQ-PCR).

RESULTSThe expression of CK19 mRNA in the cancerous tissues was 1.85 and 1.66 times higher than that in normal oral mucosa and in para-cancerous tissues, respectively. The expression of CK19 mRNA in lymph nodes from 9 patients with OSCC was positive and the positive rate was 45% (9/20). The positive rate of CK19 mRNA in all lymph nodes from 9 patients with OSCC was 81.8% (18/22), and the positive rate of CK19 mRNA in all lymph nodes from 20 patients with OSCC was 41.9%(18/43). CK19 mRNA level in the cancerous tissues relative to para-cancerous tissues and normal oral mucosa of the patients whose CK19 mRNA expression was positive was lower than that of the patients whose CK19 mRNA expression was negative in lymph nodes, respectively.

CONCLUSIONThe possible reason that the expression of CK19 mRNA in the cancerous tissues was higher than that in para-cancerous tissues and normal oral mucosa was that the CK19 synthesis in cancerous tissues increased obviously. The detection of CK19 mRNA in lymph nodes was regarded probably as one of the markers for detecting OSCC micrometastasis in lymph nodes. The detection of CK19 mRNA in lymph nodes by FQ-PCR was more sensitive than hematoxylin-eosin staining in diagnosing OSCC micrometastasis.

Carcinoma, Squamous Cell ; Female ; Humans ; Keratin-19 ; Male ; Middle Aged ; Mouth Mucosa ; Mouth Neoplasms ; RNA, Messenger