Acetates ; blood ; Adult ; Aged ; Chromatography, Gas ; methods ; Female ; Humans

Objective With the method of detecting hepatitis B virus deoxyribonucleic acid(HBV-DNA) gene in breast milk,in ordered to diagnose whether there is HBV in breast milk in those mothers whose HBsAg are positive.Methods Collected breast milk from 187 mothers whose HBsAg were positive,and complified HBV-N gene in breast milk by Nested-Polymerase chain reaction (Nested-PCR).Results Nested-PCR could detect HBV-DNA in breast milk,and the positive rate was 3.2%.Conclusion The method of detecting HBV-N gene in breast milk by Nested-PCR can detect HBV-DNA in breast milk,and it can be an laboratory evidence for whether breast feeding or not.

Objective:Aldose reductase,involved in the pathogenesis of diabetic complications,was recombinated with an adeno associated virus vector pSNAV2.0,and it was transfected into human embryonic kidney 293(HEK 293)cells.The gene engineering produced AR would be used as a target protein to screen aldose reductase inhibitors.Restriction endonuclease digestion and ligation procedures were performed to construct the AR expression plasmid vector pSNAV-hAR.Methods:After confirmation the recombinant plasmid by PCR,restriction endonuclease digestion,and DNA sequencing,pSNAV-hAR was transfected into HEK293 cells.Western blot and immunofluorescence analysis were performed to detect the expression of AR and its enzyme activity.Results:The results of a series of analysis including AR activity assay,Western blot and immunofluorescence analysis shown the expressed protein mediated by the adeno associated virus vector transfecting HEK 293 cells,was functional AR.The traditional aldose reductase inhibitors,Sobinil and Zopolrestat,were used to test and verify the constructed cell model.Conclusion:The established AR expression model can be used in mechanismresearch of activation of polyol pathway on diabetic complications and screening potential aldose reductase inhibitors.

OBJECTIVETo observe the clinical efficacy difference between segmentation scraping and conventional acupuncture for cervical spondylosis (CS) so as to provide effective treatment method.

METHODSEighty-five cases of cervical type of CS were randomly divided into a scraping group (44 cases) and an acupuncture group (41 cases). The segmentation scraping therapy was used in the scraping group. The scraping group was treated with focusing on scraping the head and joint part of neck and occiput in the upper cervical spine injury, and focusing on scraping the lower section of cervical and shoulder in the lower cervical spine injury, once every seven days, totally for 3 times. In the acupuncture group, Fengchi (GB 20),Wangu (TE 5), Tianzhu (BL 10),Neck-Jiaji (EX-B 2), etc. were selected,once daily,for 15 days. The visual analogue scale (VAS) was used to evaluate the immediate analgesic effect after the first treatment and the clinical efficacy was observed after the end of treatment.

RESULTSAfter the first treatment, the score of VAS was decreased significantly in the scaping group (P < 0.01), but there was no significant difference in the acupuncture group compared with those before treatment (P > 0.05); the score of VAS in the scaping group after the first treatment was lower than that in the acupuncture group (3.66 +/- 0.74 vs 5.43 +/- 0.35, P < 0.01). Compared with before treatment, the scores of VAS were decreased significantly after treatment in two groups (both P < 0.01), but without significant difference between two groups (P > 0.05); the effective rate was 95.5% (42/44) in the scaping group and 87.8% (36/41) in the acupuncture group, the curative effects were similar (P > 0.05).

CONCLUSIONBoth of scraping and acupuncture therapies have good analgesic effect for cervical spondylosis, and overall effects are similar, but the immediate analgesic effect of scraping thrapy is better than that of conventional acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Neck Pain ; therapy ; Spondylosis ; therapy ; Treatment Outcome

Objective:To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-in- duced apoptosis of the prostate cancer cell line DU145.Methods:The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay;cell cycle arrest was detected by flow cytometry assay;morphological change was observed by Giemsa staining;and defined kinase protein levels were determined by Western blot analysis.Re- suits:FK228 obviously inhibited DU145 cells growth,arrested cell cycle at G_0/G_1 phase,induced cells morphological changes and degraded several kinase proteins,including EGFR,Her2,Raf-1,Src,Cdk4 and IAP member Survivin.The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways,inducing apoptosis.Condu- sion:FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival sig- nal pathways of DU145 cells.

OBJECTIVETo evaluate the safety of iodine-125 seed implantation in the liver.

METHODSTwenty New Zealand rabbits were divided into control and treatment groups and in the latter, iodine-125 seeds of 37 MBq were implanted into the liver under CT guidance whereas nonradioactive seeds were implanted in the control rabbits. Four weeks after implantation, white blood cell count, liver functions, and renal functions were measured or evaluated for comparison with those before implantation. The rabbits were then anesthetized to collect the liver tissue for pathological examination with HE staining and cell apoptosis assay.

RESULTSObvious hepatic tissue necrosis was observed around the radioactive seeds in the treatment group. At a 5 mm distance to the seeds, a distinct boundary occurred between the necrotic hepatic cells and normal cells. The control rabbits, however, had normal liver structure around the seeds implanted. In situ cell apoptosis examination showed a distinct band of apoptotic cells in the liver tissue of rabbits in the treatment group, which was not found in the control group. Two weeks after iodine-125 irradiation, alanine aminotransferase significantly increased in the treatment group (t=6.285, P<0.001), but recovered two weeks later (t=2.002, P=0.06). No significant alterations occurred in aspartate aminotransferase, blood urea nitrogen, serum creatinine, hemoglobin, serum total bilirubin, white blood cell count, or platelet count after the seed implantation.

CONCLUSIONIodine-125 seed implantation in the liver results in conformal irradiation dose distribution without obvious effects on the vital organs, demonstrating iodine-125 seed implantation as a safe and minimally invasive technique for hepatic cancer treatment.

Alanine Transaminase ; blood ; Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; In Situ Nick-End Labeling ; Iodine Radioisotopes ; adverse effects ; Liver ; pathology ; physiopathology ; radiation effects ; Male ; Rabbits ; Radiation Injuries, Experimental ; blood ; etiology ; pathology ; Random Allocation ; Time Factors

Objective:To investigate the prevalence of enterovirus infection and its clinical characteristics in neonatal febrile cases.Methods:A total of 308 neonates in the neonatal ward of Kunming Children′s Hospital with febrile symptoms from March 2018 to February 2019 were selected for the study.Fecal specimens and some (271) cerebrospinal fluid specimens were collected from all neonates.Enterovirus was respectively detected in feces and cerebrospinal fluid by using reverse transcription-polymerase chain reaction (RT-PCR) method.The rate of enterovirus infection in febrile neonates was clarified.They were divided into enterovirus infection group ( n=91) and no enterovirus infection group ( n=217). The clinical data of the subjected neonates in the two groups were analyzed and the clinical manifestations of the neonates and their laboratory findings were compared statistically. Results:Ninety-one enteroviruses (90 enterovirus were general type, and 1 was coxsackievirus A16 type) were detected in 308 children, accounting for 29.55% of all neonates.Viral RNA was detected in only 1 case [0.37% (1/271)]of all examined cerebrospinal fluid specimens, which was significantly lower than the detection rate of 29.55% (91/308) in fecal specimens.There were no fatal cases, but there was one severe case in enterovirus infection group.Except for the severe case which was discharged automatically, all of them improved or healed and discharged from hospital.The clinical manifestations of the enterovirus infection group were non-specific statistically compared with the non-enterovirus infection group(all P>0.05), except for respiratory symptoms and skin rash(all P<0.05). The incidence of enterovirus infection was significantly seasonal[83.52%(76/91)in summer and autumn]. Conclusion:In the epidemic season of enterovirus, it is recommended to list fecal enterovirus as a routine test item to improve the accuracy and timeliness of diagnosis and treatment, and avoid the application of unnecessary antibiotics and the outbreak of enterovirus in the neonatal ward.

OBJECTIVETo study the apoptotic effect and mechanisms of methylmercury (MeHg) on HL-7702 cell line in vitro.

METHODSIn this study, the cell apoptosis was observed by AO/EB method and FCM method; the mitochondrial membrane potential was detected by FCM; and the expression of proteins related to apoptosis was measured by immunocytochemical method.

RESULTSAfter exposure to MeHg for 24 h in different doses, apoptotic rate ascended with the increasing of MeHg concentration. By AO/EB method, cell apoptotic ratio of negative control group was (2.62 +/- 0.19)%, cell apoptotic ratio of 10-50 micromol/L exposure groups were (7.97 +/- 0.64)%, (12.66 +/- 0.76)%, (19.16 +/- 0.87)%, (18.42 +/- 0.88)%, and (11.52 +/- 0.63)%, there were significant differences between the exposure and negative control groups (q values were 17.057, 32.009, 52.732, 50.373, 28.375; P<0.05). Mitochondrial membrane potential descended with the increase of MeHg, mitochondrial membrane potential of negative control group was (10.23 +/- 3.43) mV, mitochondrial membrane potential of 10-50 micromol/L exposure groups were (3.25 +/- 0.66), (3.03 +/- 0.35), (1.68 +/- 1.26), (1.69 +/- 1.13) and (1.77 +/- 0.88) mV, and there was significant differences between exposure and negative control groups (q values were 9.569, 9.871, 11.722, 11.708, 11.598; P<0.05). The expression of Bax, Bcl-2, CytC, Caspase-3 and AIF enhanced with the increase of MeHg, Bax/Bcl-2 ratio also appeared a trend of increase. Bax expression integral optical density (IOD) of negative control group was (21295.86 +/- 1969.81), Bax expression IOD of 10, 20, 30 micromol/L groups were 42807.87 +/- 4416.64, 55651.65 +/- 4662.72, and 72708.56 +/- 910.10, there were significant differences in Bax expression between 10, 20, 30 micromol/L groups and negative control group (q values were 14.191, 14.320, 33.917; P<0.05); Bcl-2 expression IOD of negative control group was (12588.33 +/- 4091.02), Bcl-2 expression IOD of 10, 20, 30 micromol/L groups were 20539.16 +/- 4906.09, 23689.97 +/- 2281.42, and 28692.80 +/- 4655.86, there were significant differences in Bcl-2 expression between 10, 20, 30 micromol/L groups and negative control group (q values were 4.322, 6.035, 8.754; P<0.05); and AIF expression IOD of negative control group was (12942.72 +/- 457.94), AIF expression IOD of 10, 20, 30, 40 micromol/L groups were 16973.57 +/- 1922.87, 29998.91 +/- 6803.58, 52467.16 +/- 1916.25 and 106342.53 +/- 1273.19, there were significant differences in AIF expression between 20, 30 and 40 micromol/L groups and negative control group (q values were 11.449, 26.530, 62.692; P<0.05).

CONCLUSIONMeHg could induce apoptosis on HL-7702 cell line in vitro. The mechanisms could be related to mitochondrial pathway in apoptosis.

Apoptosis ; drug effects ; Cell Line ; Flow Cytometry ; Hepatocytes ; cytology ; drug effects ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; Methylmercury Compounds ; pharmacology ; Mitochondrial Proteins ; metabolism

OBJECTIVETo investigate the changes of ataxia-telangiectasia mutated (ATM) phosphorylation in HepG(2) cells in relation to HepG(2) cell survival under continuous low dose-rate irradiation.

METHODSHepG(2) cells were exposed to equivalent irradiation doses delivered at either a continuous low dose-rate (7.76 cGy/h) or a high dose-rate (4500 cGy/h), and the phosphorylated ATM proteins and surviving fraction of HepG(2) cells after the exposures were compared.

RESULTSThe phosphorylation of ATM protein was maximal at 0.5 Gy irradiation delivered at either a high doserate or a continuous low doserate. As the radiation dose increased, ATM protein phosphorylation decreased under continuous low dose-rate irradiation, but remained stable under high dose-rate irradiation. With comparable ATM protein phosphorylation induced by continuous low dose-rate irradiation and high dose-rate irradiation, there was no significant difference in the surviving fraction of HepG(2) cells (P>0.05), but at a significantly lower ATM protein phosphorylation level than that induced by high dose-rate irradiation, continuous low dose-rate irradiation resulted in increased cell killing (P<0.01).

CONCLUSIONContinuous low dose-rate irradiation increases HepG(2) cells radiosensitivity as compared with high dose-rate irradiation. Increased cell killing following continuous low dose-rate irradiation is associated with reduced phosphorylated ATM protein, and inhibition of ATM phosphorylation may increase the radiosensitivity of HepG(2) cells.

Animals ; Ataxia Telangiectasia Mutated Proteins ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cell Survival ; radiation effects ; DNA-Binding Proteins ; metabolism ; Dose-Response Relationship, Radiation ; Humans ; Mice ; Phosphorylation ; radiation effects ; Protein-Serine-Threonine Kinases ; metabolism ; Radiation Tolerance ; radiation effects ; Time Factors ; Tumor Suppressor Proteins ; metabolism

The effects of adrenomedullin (ADM) on the L-type calcium currents (I(Ca,L)) and the mechanism of the signal transduction process were studied. Enzymatically isolated guinea-pig ventricular myocytes were used to measure ICa,L with whole-cell patch-clamp techniques. ADM at the concentrations of 1-100 nmol/L decreased ICa,L in a dose-dependent manner (P<0.05). ADM22-52) (100 nmol/L), a specific ADM-receptor antagonist, completely abolished the ADM-induced inhibition of ICa,L. Pretreatment of the cells with H-89 (10 micromol/L), a specific PKA inhibitor, did not attenuate the effects of ADM. Intracellular application of 10 micromol/L PKC19-36), a specific PKC inhibitor, prevented the ADM-induced inhibition of the ICa,L, while the specific PKC activator PMA could mimic the effects of ADM on the ICa,L. PMA (1 micromol/L) decreased the ICa,L by 32.26+/-4.20%(P<0.05). These findings indicate that ADM can inhibit the ICa,L in guinea-pig ventricular myocytes, and the inhibition is mediated by the specific ADM-receptor and an activation of protein kinase C.
Adrenomedullin ; pharmacology ; Animals ; Calcium Channels, L-Type ; metabolism ; Guinea Pigs ; Heart Ventricles ; cytology ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Protein Kinase C ; metabolism