Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.

OBJECTIVE: To develop a DNA detection kit for identification of Fetus Cervi, evaluate the quality indexes including specificity, stability, sensitivity and repeatability, and inspect the qualities of commercial Fetus Cervi samples. METHODS: The Fetus Cervi DNA test kit was developed and modified using the classical DNA extraction and PCR identification method. The genomic DNAs of Fetus Cervi and counterfeit goods were extracted by the kit and PCR technique was performed to identify the authenticity. The purity of DNA was detected by UV spectrophotometer. Finally, commercially available Fetus Cervi samples were verified. RESULTS: The kit was proved to be effective after 20 times of repeated frozen-thawing and it could be stored at - 20℃ for 1 year. The DNA could be detected when the primary solution was diluted by 200 times. The specificity test confirmed that the 15 samples were authentic Fetus Cervi, and 7 were counterfeit. The specificity of the kit was 100%. The repetitive experiments showed that the average intra-assay CV% and inter-assay CV% of the kit were 2.32% and 2.56%, respectively. CONCLUSION: The amount and purity of the nucleic acids extracted by the DNA detection kit can meet the requirement for identification of Fetus Cervi, and the kit has good specificity, high sensitivity and good stability, so it is suitable for the rapid detection of Fetus Cervi.

OBJECTIVE: To explore a practical method for isolation of mitochondrial DNA from Fetus Cervi and set up an accurate method to distinguish Fetus Cervi from adulterants. METHODS: The improved SDS alkaline lysis and salt-outing METHODS were used to extract mtDNA and genomics DNA from Fetus Cervi and fetus from adulterant animals respectively. A set of primers were designed by bioinformatics to establish PCR system and reaction which could be used to identify Fetus Cervi species. RESULTS: The mtDNA extracted from Fetus Cervi by improved SDS alkaline lysis method had high content and purity, which met the PCR response requirements. There was a pair of primers which could distinguish Fetus Cervi from adulterants. CONCLUSION: Extraction of mtDNA by improved SDS alkaline lysis method is practical and accurate. The distinctive PCR can distinguish Fetus Cervi from adulterants and counterfeit animal fetus and it is accurate and reliable.

Objective To evaluate the effect of comprehensive prevention and control of soil?transmitted nematodiasis in Runzhou District Zhenjiang City Jiangsu Province from 1997 to 2012. Methods The comprehensive prevention and control measures included the helminthicide health education improvement of water supplier and harmless toilets and these mea?sures were implemented continuously. At the same time the infection rates of soil?transmitted nematodes the local economic in?dicators and the coverage rates of tap water and harmless toilets were surveyed. Results The mass chemotherapy was per?formed for 202 100 person?times and the diagnosed chemotherapy was performed for 2 006 person?times in Runzhou District from 1997 to 2012. The awareness rates of health knowledge were 57.18%in 1997 and 95.62%in 2012. The coverage rates of tap water and harmless toilets were 10.14%and 10.21%in 1997 and 100.0%and 90.38%in 2012 respectively. There were negative correlations between the awareness rate of per capita GDP per capita net income coverage rates of tap water cover?age rates of harmless toilets health knowledge and the infection rate of soil?transmitted nematodes respectively rper capita GDP=-0.526 P 0.05 rper capita net income=-0.671 P 0.01 rcoverage rates of tap water=-0.936 P 0.01 rcoverage rates of harmles toilets=-0.922 P 0.01 rawarenes=-0.774 P 0.01 . The statistical analysis showed that the infection rate of soil?transmitted nematodes had a downward trend as an expo?nential curve in Runzhou District from 1997 to 2012 y=42.031 7e?0.357 6x R2=0.803 6 F=57.28 P=0.000 . The infection rate of degradation by an annual rate was 29.18%. The infection rate in farmers was significantly higher than that in students χ2=17.998 P 0.01 . There was no significant difference between men and women in the infection rate of soil?transmitted nema?todes χ2=3.627 P=0.057 . Conclusion The comprehensive prevention and control measures and the development of so?cial economy contribute to the steady decline of soil?transmitted nematode infections.

The strain Acidiphilium cryptum DX1-1 producing PHB was irradiated respectively by UV and Co60 to raise PHB production. The results indicated that the effect of UV better than using Co60. One strain of the UV mutagenized called UV60-3 has the highest PHB production yield, showing final PHB concentra- tion of 28.56 g/L, 1.45 times higher than that of original strain. FT-IR spectroscopy analysis shows that the polymers obtained from the strain DX1-1 have the same IR spectra of standard PHB. Further research about the best appropriate C/N ratio of the mutant was done. The optimum ratio of C/N was about 3.76, the final PHB concentration reaches to 30.57 g/L.

OBJECTIVEThis study sought to analyze the clinical manifestations and intervention of fulminant septic shock in community-acquired Pseudomonas aeruginosa septicemia.

METHODSWe retrospectively reviewed the medical records for diagnosis, antibiotic therapy, clinical course of septic shock, respiratory support, laboratory data etc.

RESULTSEight of nine cases with P. aeruginosa septic shock died. Fever (nine cases) and cough (three cases) or diarrhea (3 cases) were the 2 most common initial symptoms, three cases developed skin gangrenosum later. Pseudomonas aeruginosa infection was not considered in any of the cases before death or blood culture showed positive result. Only 3 cases were initially treated with susceptible antibiotic regimen but no anti pseudomonas combination therapy was applied, susceptible antibiotic monotherapy was applied in 7 cases after transfer to the ICU. The mean latency of shock occurrence was 5.1 hours (range 0 to 21 hours) after admission, the mean duration from the occurrence of shock to death was 13.8 hours (range, 1 - 32 hours). All the patients were transfer red to ICU for shock, the appropriate resuscitation of shock patients was delayed by 49.3 minutes (range 25 - 80 minutes) by transfer. Only two cases were diagnosed and treated for shock on admission; after transferred to ICU, only 5 patients were diagnosed as having shock, and only 3 received anti-shock treatment. Eight of the patients died of persistent shock. In 6 patients who died, mechanical ventilation was not applied until cardiac arrest occurred. All the patients had hypoalbuminaemia, elevated serum C-reactive protein concentration, leukopenia and 6 cases had DIC.

CONCLUSIONThe initial presentation of the cases with community-acquired Pseudomonas aeruginosa septicemia was nonspecific with fever and cough or diarrhea. Clinicians often underestimated the severity of the infection, few patients received effective antimicrobial therapy. The authors suggest that an anti-pseudomonas antibiotic should be included in the initial empiric antibiotic regimen to cover P. aeruginosa high-risk patients; the front-line clinician should be educated for early recognition and aggressive resuscitation of P infection. aeruginosa septicemia.

Adolescent ; Child, Preschool ; Community-Acquired Infections ; Female ; Humans ; Infant ; Male ; Pseudomonas Infections ; Pseudomonas aeruginosa ; Retrospective Studies ; Shock, Septic ; microbiology

OBJECTIVETo investigate the changes in cisplatin sensitivity of resistant ovarian cancer A2780 cells after inhibition of miR-23a expression and explore the molecular mechanisms.

METHODSThe drug-resistant ovarian cancer A2780 cells were exposed to cisplatin alone or in combination with antagomir-23a. The cell inhibition rates after the treatments were detected using MTT assay, cell cycle changes assessed with flow cytometry; and apoptotic cells observed using Hoechst33258 staining. The changes in glycoprotein P-gp expression in the cells were detected using Western blotting.

RESULTSInhibition of miR-23 a combined with cisplatin treatment significantly increased the cell inhibition rate (P<0.01) and lowered the IC(50) so of cisplatin by 83.76% from 110.18 μmol/L in the control group to 17.89 μmol/L (P<0.01). The combined treatments also caused cell cycle arrestin G0/G1 phase, increased the cell apoptosis rate (P<0.01) and the number of cells stained with Hoechst33258; the cellular expression of P-gp protein was significantly reduced as the cisplatin doses increased (P<0.01).

CONCLUSIONInhibition of miR-23a expression increases the sensitivity of A2780 cells to cisplatin possibly by inhibiting the negative regulation by miR-23a target genes that causes inhibition of P-gp protein expression.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Humans ; MicroRNAs ; metabolism ; Ovarian Neoplasms ; pathology

OBJECTIVETo explore the effect of Jiangtang Xiaozhi capsule (JXC) on morphological changes of islets and liver at rat model of type 2 diabetic mellitus and provide the experimental basis for the clinical therapy of type 2 diabetic mellitus.

METHODWister rats were fed on a diet enriched in fat and glucose to induce insulin resistan, the rats were injected intrapertoneally with a low-dose streptozotocin (STZ) twice (25 mg x kg(-1)) to induce hyperglycemia, so the successful rat model of type 2 diabetes were established. The experimental rats were divided into model group, high dose JXC group, middle dose JXC group, low dose JXC group, Erjiashuanggua group, Jinqijiangtang group and normal control group. After all the treatment groups received their own medicine for two months, all the rats were sacrificed and morphological examination on their islets and livers were performed.

RESULTFatty liver in various degrees was seen in the model group and all the treatment groups, but the liver steatosis in middle and low dose JXC groups was significantly milder than that in model group (P < 0.05). Islets in the high dose JXC group were significantly more than that in the model group (P < 0.05).

CONCLUSIONJXC can improve significantly the pathological change in islets and liver steatosis at rat model of type 2 diabetic mellitus.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Islets of Langerhans ; drug effects ; pathology ; Liver ; drug effects ; pathology ; Male ; Microscopy ; Rats ; Rats, Wistar

This study aimed to investigate the reconstruction of the thumb and finger extension function in patients with middle and lower trunk root avulsion injuries of the brachial plexus.From April 2010 to January 2015,we enrolled in this study 4 patients diagnosed with middle and lower trunk root avulsion injuries of the brachial plexus via imaging tests,electrophysiological examinations,and clinical confirmation.Muscular branches of the radial nerve,which innervate the supinator in the forearm,were transposed to the posterior interosseous nerve to reconstruct the thumb and finger extension function.Electrophysiological findings and muscle strength of the extensor pollicis longus and extensor digitorum communis,as well as the distance between the thumb tip and index finger tip,were monitored.All patients were followed up for 24 to 30 months,with an average of 27.5 months.Motor unit potentials (MUP) of the extensor digitorum communis appeared at an average of 3.8 months,while MUP of the extensor pollicis longus appeared at an average of 7 months.Compound muscle action potential (CMAP) appeared at an average of 9 months in the extensor digitorum communis,and 12 months in the extensor pollicis longus.Furthermore,the muscle strength of the extensor pollicis longus and extensor digitorum communis both reached grade Ⅲ at 21 months.Lastly,the average distance between the thumb tip and index finger tip was 8.8 cm at 21 months.In conclusion,for patients with middle and lower trunk injuries of the brachial plexus,transposition of the muscular branches of the radial nerve innervating the supinator to the posterior interosseous nerve for the reconstruction of thumb and finger extension function is practicable and feasible.