Objective To investigate the efficiency of three methods to detect HPV infection of desquamated cervical epithelial cells and the relationship between methods and TCT.Methods hC2 method was used in disconnecting the HPV DNA chain in the samples to hybrid captures,and examine the 13 types of HPV which were highly related to cervical carcinoma.Q-PCR was first used to extract HPV DNA of samples and expanded nucleotide to be examined on Q-PCR to obtain results directly.MPHC was used to extract HPV DNA and to amplify nucleotide.The products was added to micro-well plate to proceed hybridization,and enzyme label was added,and results were studied in the enzyme labelling apparatus.The results of the latter two methods were the same as hC2.hC2 matched Q-PCR was used to examine 260 eases of desquamated cells from HPV infection against 200 cases of desquamated cells from HPV infection,and the results were compared by using MPHC matching examination.Results 1.With three methods,HPV infection was found to be 100% in all the abnormal cervical squamous epithelial cells.2.The sensitivity of the methods,howerer,was different.The QFCR and MPHC had higher positive rate than hC2.Positive HPV infection rate with Q-PCR and MPHC was the same as that of hC2 in the majority of cases,as in a few cases,it was positive with Q-PCR and MPHC but negative with hC2 alone False positive rate was 2% in MPHC.Conclusion HPV testing combined with the TCT routine is the best screening program for detecting cervical cancer and precursor of cervical careinoma.

AIM: To study the effects of disulfiram/copper complex (DSF/Cu) on ultrastructures and mechan-ical properties of human breast cancer and normal breast epithelial cells by atomic force microscopy (AFM) based on the nanoscale resolution and piconewton force measurement level.METHODS: The change of cell cycle and apoptotic rate of MCF-7 cells and MCF-10A cells induced by DSF/Cu were compared by flow cytometry.The cell surface morphology, ultra-structure, height, width and roughness were detected by AFM.The effects of DSF/Cu on the hardness (Young’s modu-lus) of the 2 kinds of cells were determined by AFM with indentation technique.RESULTS: DSF/Cu significantly in-duced apoptosis of the MCF-7 cells in a dose-dependent manner, whereas had little effect on the MCF-10A cells.The cell cycle analysis showed that DSF/Cu induced G2 /M arrest in the MCF-7 cells, but led to G0 /G1 arrest in the MCF-10A cells.The AFM images showed that the MCF-7 cells shrank and showed smaller and smoother morphology, and the filopo-dia were retracted obviously, even some became into lamellipodia, or disappeared completely after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.The quantitative analysis indicated that the MCF-7 cells showed smaller width and
larger height, and the root mean square roughness and average roughness were decreased significantly in a dose-dependent manner after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.However, little effect in the MCF-10A cells was observed.The biomechanics test at a single cell level demonstrated that the Young’s modulus of the MCF-7 cells and MCF-10A cells were both increased, yet the proportion increased in the MCF-7 cells was much higher than that in the MCF-10A cells after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.CONCLUSION: DSF/Cu has strong antitumor effect on breast cancer with high efficiency and low toxicity by changing the properties of the biomechanics specifically.

AIM To explore the radioprotective function of Xihuang Capsules (Bovis Calculus artijactus,Moschus,Olibanum,Myrrha) in patients with acute radiation-induced oral mucositis and its mechanisms.METHODS Eighty patients with nasopharyngeal carcinoma undergone radiotherapy were randomized into two groups:treatment group (radiation plus Xihuang Capsules) and control group (radiation alone).The comparison of two groups in the onset and the tolerated dose of mucositis was made according to the acute radiation injury classification standard.The levels of TNF-α and IL-6 in saliva were detected by ELISA.RESULTS After the treatment with Xihuang Capsules,the onset of oral mucositis delayed significantly and the tolerated dose elevated markedly comparing with the control group (t =2.180,12.930,P ＜ 0.05).The morbidity rate of Ⅲ-N-degree of oral mucositis in the treatment group was lower than that of the control group at the dosage of 40 Gy and 70 Gy,respectively (Z =3.661,4.270,P ＜0.01).Furthermore,there was no difference in the levels of TNF-α and IL-6 in the pre-treatment in the two groups (t =1.010,1.469,P ＞ 0.05).With the increase in radiant dose,the levels of TNF-α and IL-6 in the two groups both elevated dramatically,and peaked at the dose of 40 Gy,but both levels in the treatment group were lower than those of the control group (t =8.305,6.069,P ＜ 0.05).When DT =70 Gy,the levels of TNF-α and IL-6 were lower than pre-radiation,and the levels of TNF-α and IL-6 in the treatment group were lower than those in the radiation alone group (t =3.835,2.488,P ＜ 0.05).CONCLUSION Xihuang Capsules can delay and reduce acute radiation-induced oral mucositis and improve radiation tolerated dose,so it may involve the release of TNF-oα and IL-6 in saliva.

Objective To study the combination effects of telmisartan and pioglitazone on the expression of heparanases (HPA) and podocin in the kidney of diabetic nephropathy (DN) rats and its possible mechanism. Methods DN model rats were established by intraperitoneal injection with STZ for 12 weeks. All the DN rats were randomly divided into telmisartan group (T group), pioglitazone group (P group), combination of telmisartan and pioglitazone group (L group), and DN group (D group). Healthy rats were chosen as healthy control group(N group). After garage with drugs for 12 weeks, 24-h urinary protein and serum biochemical indicators were examined. RT-PCR and immunohistochemistry methods were applied to detect the mRNA and protein expression of HPA and podocin. Results Compared with T group and P group, 24-h urinary protein of L group was markedly decreased(P<0.05). Compared with the N group, the level of fasting blood glucose, relative renal weight, BUN and Scr in other 4 groups were markedly increased (P<0.05). Compared with T group and P group, the Scr level and the expression of HPA mRNA and protein in L group was markedly decreased (P<0.05), and the protein and mRNA expression of podoein in L group was markedly increased (P<0.05). Conclusion Combination of telmisartan and pioglitazone can down-regulate the mRNA and protein expression of HPA of glomerular basement membrane and up-regulate the protein expression of podocin of podocyte in DN rats, which may ameliorate the proteinuria.

AIM:To explore the influence of exogenous hydrogen sulfide ( H2 S) on coagulation and fibrinoly-sis in ferric chloride ( FeCl3 )-induced mouse carotid artery thrombosis .METHODS: The mice were divided into sham control group, model group, different concentrations (12.5, 25 and 50μmol/kg) of sodium hydrosulfide (NaHS, H2S do-nor) groups and 30 mg/kg clopidogrel ( positive control ) group.Intraperitoneal injection of NaHS at different concentra-tions and oral administration of clopidogrel bisulfate were performed for 3 d prior to FeCl 3-induced carotid artery thrombo-sis.The frozen sections of the carotid artery were collected to perform HE staining , and the thrombus pattern and the chan-ges of vascular pathology were observed .The thrombus was weighed to calculate thrombosis inhibitory rate .Prothrombin time ( PT) , activated partial thromboplastin time ( APTT) , fibrinogen ( FIB) and fibrinogen degradation product ( FDP) in the mice were also measured by a coagulometer .The plasma levels of thromboxane B 2 ( TXB2 ) , 6-keto-prostaglandin F 1α(6-keto-PGF1α) and plasminogen activator inhibitor (PAI) were detected by ELISA.RESULTS: Compared with model group, NaHS dose-dependently inhibited the formation of carotid artery thrombus .NaHS treatment reduced the contents of TXB2 and PAI, and recovered 6-keto-PGF1αcontent in thrombosis model group .In NaHS treatment groups , 6-keto-PGF1α/TXB2 and thrombus weight was negatively correlated .NaHS treatment prolonged PT and APTT , reduced the content of FIB, but increased the level of FDP in thrombosis model group .CONCLUSION:Hydrogen sulfide prevents FeCl 3-induced carotid artery thrombosis by inhibiting coagulation and activating fibrinolysis .

OBJECTIVE: To determine whether the composite acoustic therapy is effective to treat tinnitus in patients with sudden deafness and to explore the mechanisms. METHOD: Ninety-six cases (96 ears) were divided into experimental group and control group, and all the patients underwent drug treatment. The patients in experimental group were given personalized composite acoustic therapy in the first 30 days, music therapy in next 31-90 days, however, the patients in control group were not given sound therapy. Additionally, pure tone audiogram, tinnitus pitch and loudness as well as questionnaires (including THI, VAS, and SAS) were conducted for each patient before treatment, at day 30 and day 90 posttreatment. RESULT: Eighty-nine patients (n = 47 for experimental group and n = 42 for control group) completed the trial. The results of day 30 posttreatment showed there were no significant differences in VAS and hearing recovery rate between these two groups, but THI and SAS showed significant differences. The results of day 90 posttreatment showed significant differences in VAS (P < 0.05), THI (P < 0.01) and SAS (P < 0.01), and no significant difference of hearing recovery rate was detected. The most significant changes of VAS appeared in the first 30 days of treatment. The hearing and tinnitus recovery in experimental group were better for the first 30 days of treatment than for the next 31-90 days, and similar results were obtaind in control group. (P < 0.01). CONCLUSION Composite acoustic treatment combined with drug therapy can improve the tinnitus and anxiety symptoms of patients with sudden deafness, and the effect on hearing recovery still need to be confirmed in further studies.
Acoustics ; Anxiety ; Hearing Loss, Sudden ; complications ; therapy ; Hearing Tests ; Humans ; Surveys and Questionnaires ; Tinnitus ; complications ; therapy ; Treatment Outcome

Objective To investigate the protective effect of breviscapine on ischemia-reperfusion renal injury, which provides scientific theoretical basis for clinical prevention and treatment of renal ischemia-reperfusion injury. Method 36 SPF male healthy SD rats were randomly divided into 3 groups, 12 rats in each group. Group A was ischemia-reperfusion group, group B was erigeron breviscapus injection preconditioning group, and group C was sham operation group. Rats in group A and C were injected with normal saline, while rats in group B were given 12 ml/kg erigeron breviscapus injection by intraperitoneal injection., After 14 days, renal function, renal antioxidant indexes, renal cell apoptosis indexes and expression of Bcl-2, Bax in renal tissue of three groups were compared. Results The indexes of renal function showed that blood urea nitrogen (BUN) and Serum creatinine (SCr) in group B were significantly lower than group A (P<0.05), but significantly higher than group C (P<0.05). Renal antioxidant indexes showed that superoxide dismutase (SOD) and malondialdehyde (MDA) in group B were significantly higher than group A (P<0.05), but significantly lower than group C(P<0.05). The index of renal cell apoptosis showed that the apoptosis index AI in group B was significantly lower than group A (P<0.01), but significantly higher than group C (P<0.01). The results of immunohistochemical staining showed that the expression of Bcl-2 in group B were significantly higher than that of group A and group C (P<0.05), and the expression of Bax in group B was significantly lower than that in group A (P<0.05) while significantly higher than group C (P<0.01). The ratio of Bcl-2/Bax in group B was significantly higher than group A and group C (P<0.05). Conclusion Erigeron Breviscapus Injection may have a protective role on renal ischemia reperfusion injury though antioxidant and anti-apoptosis.

AIM: To investigate the role of chloride channels in the apoptosis of human poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As2O3).METHODS: The apoptotic rates of CNE-2Z cells induced by As2O3 for 24 h or 48 h were monitored by flow cytometry.The technique of whole-cell patch clamp was used to record the currents activated by As2O3 in the CNE-2Z cells.The inhibition of As2O3-induced apoptosis by chloride channel blocker DIDS in the CNE-2Z cells was analyzed by flow cytometry.RESULTS: As2O3 at 5 μmol/L induced apoptosis of CNE-2Z cells in time-dependent manner.The currents with outward rectification were activated when the cells were exposed to 5 μmol/L As2O3.No obvious time-and voltage-dependent inactivation of the currents was observed.The reverse potential of the currents was close to the equilibrium potential for chloride.The activated currents were inhibited by the chloride channel blockers NPPB and DIDS.The 47% hypertonic solution inhibited the activated currents completely.Chloride channel blocker DIDS inhibited the apoptosis of CNE-2Z cells induced by As2O3.CONCLUSION: As2O3 activates volume-sensitive chloride channels, and chloride channels may play an important role in the apoptosis of CNE-2Z cells induced by As2O3.

Aim To study the mechanism of DSF-Cu induced apoptosis of human nasopharyngeal carcinoma CNE-2 Z cells by affecting the function of mitochondria and cytoskeleton. Methods The cell cycle,the rate of apotosis,the levels of intracellular ROS and MMP in CNE-2 Z cells were tested by flow cytometry after trea-ted with different concentration of DSF-Cu. The chan-ges of the cell surface morphology, ultrastructure, cell height, width and roughness were detected by AFM. The distribution and reorganization of cytoskeleton F-actin were observed by Laser scanning confocal micro-scope. Results Cells were incubated with different concentration of DSF-Cu ( 0 ~200 nmol · L-1 ) for 24 h, the apoptotic ratio increased significantly and the treatment of DSF-Cu resulted in a concentration-de-pendent accumulation of CNE-2Z cells in G2/M phase. Furthermore,the treatment of DSF-Cu was able to in-crease the production of intracellular ROS and decrease the MMP in CNE-2Z cells. In addition,AFM imaging showed that compared to the control group,with the in-crease of DSF-Cu concentration,the CNE-2Z cells be-came smaller, cytoplasm condensed, the height in-creased,and the surface roughness reduced. Moreover, the filopodia became shorter, shrinked and even com-pletely destroyed after treated with different concentra-tion of DSF-Cu. At last,the LSCM image showed that the fluorescence intensity of F-actin networks was de-creased, then the structure was rearranged and de-stroyed obviously by treated with DSF-Cu. Conclusion DSF-Cu can induce apoptosis and arrest cell cycle at G2/M phase in CNE-2Z cell through a mitochondria-dependent pathway. Above findings highlight the appli-cations of AFM at the single cell level for the investiga-tion of antineoplastic drug in nasopharyngeal carcinoma therapy.

[ ABSTRACT] AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein 3 ( ClC-3) gene on bones of mice .METHODS: The tail gene detection assay was used to confirm the overexpression of ClC-3.The male FVB mice of three months old were divided into two groups , the wild type ( WT) group and the ClC-3 overexpressed (ClC-3 transgene) group.The body weight, length and weight of the right tibias were measured .The upper and middle parts of the tibias were dissected , decalcified, paraffin-imbed, sectioned and stained with HE staining .The bone morphology metrology was used to analyze the changes of bone structures .The percent trabecular area (%Tb.Ar), trabecular number ( Tb.N) , trabecular width ( Tb.Wi) and trabecular separation ( Tb.Sp) of cancellous bone in the upper part of the tibia were measured.The total tissue area (T.Ar), cortical area (Ct.Ar), percent cortical area (%Ct.Ar), marrow area ( Ma.Ar) and percent marrow area (%Ma.Ar) of the cortical bone in the middle part of the tibia were detec-ted .RESULTS:The wild type mice and the ClC-3-overexpressed mice were verified by the tail gene detection assay . Compared with WT group , the body weight and the length and weight of the tibia were decreased in ClC -3 transgene mice (P<0.05).In the cancellous bones of ClC-3 transgene mice, the%Tb.Ar and Tb.Wi were decreased (P<0.05), the Tb.Sp was increased (P<0.05) and the Tb.N was not significantly changed .In the cortical bones of ClC-3 transgene mice, the T.Ar, Ct.Ar and%Ct.Ar were decreased (P<0.05), the%Ma.Ar was increased (P<0.05), and the Ma. Ar was not significantly changed .CONCLUSION:ClC-3 overexpression may lead to the reduction of the bone mass and the destructure of the cancellous and cortical bones .The results suggest that ClC-3 may be involved in the regulation of bone resorption and/or formation.