OBJECTIVE To assess the outcomesof three different therapeutic approaches for the treatment of T3N0M0(stage Ⅲ) glottic squamous cell carcinoma. METHODS Sixty-five cases of T3N0M0 glottic squamous cell carcinoma treated with curative intent by three different modalities include vertical hemilaryngectomy(VHL,n=21),total laryngectomy(TL,n=31)and radiotherapy(RT,n=13) were reviewed retrospectively. The survival rate,recurrence rate at the primary lesion site and jugular lymph node, and laryngeal preservation was compared among three methods. RESULTS There was no significant difference in the overall survival rates(Cox proportion hazard model) and recurrence rates at the primary lesion site or jugular lymph node among the three methods. Laryngeal function was preserved in 100% of the cases in the VHL and RT groups after initial treatment. CONCLUSION The three treatment modalities had statistically similar survival and recurrence rates. Patients treated with VHL and RT had a higher rate of laryngeal preservation compared to that of TL, hence VHL or RT is a valid alternative to TL in treating selected patients with T3N0M0 glottic squamous cell carcinoma.

Objective To compare the result differences of different dynamic nerves transferring to different segments of the radial nerve. Methods From 1997 to 2000, different ways of intercostal nerves or contralataral C 7 transfering to different segments of the radial nerves were carried out in 36 cases with total avulsed brachial plexus injuries that were followed up for average 42.39 months. Four-fold table was selected to statistically calculate the effective rate of muscle strength recovery. Results The total effective rate was 56% (20/36). The effective rate of the functional rehabilitation of elbow extension was 5/8 in the group with neurotization to radial nerve trunk and 4/4 in the group with neurotization to branches innervating the triceps brachii muscle of radial nerve. The highest effective rate up to 75% (9/12) of the functional rehabilitation of wrist and digit extension belonged to the group with contralateral C 7 transferring to the radial nerve segment in the humeral spiral groove where branches innervating the triceps brachii muscle had already diverged. Conclusions The way of neurotization to radial nerve trunk and to branches innervating the triceps brachii muscle of the radial nerve can win good functional rehabilitation of elbow extension. Contralataral C 7 transfering to the radial nerve segment in the humeral spiral groove ranks the best way for function recovery of the wrist extension. Age below 30 years and operation interval less than 12 months are helpful for functional rehabilitation of the radial nerve.

Objective:To investigate the effect of heat shock protein B8 (HspB8) downregulation on retinal ganglion cell (RGC) and retinal function in the mice model of optic nerve injury (ONC).Methods:Adeno-Associated Virus (AAV) 2 AAV2-shHspB8-GFP was constructed to knockdown HspB8. 66 adult male C57/BL6 mice were randomly divided into the control group, the ONC group, the AAV2-shHspB8 group, the ONC+AAV2-shHspB8 group, and the ONC+AAV2-GFP group. There were 10, 20, 16, 10 and 10 mice respectively, and both eyes were used as experimental eyes. Western blot was used to evaluate the expression of HspB8 on day 3 and 7 after ONC. By GFP immunofluorescence staining, the efficacy of AAV2-shHspB8-GFP transfer was accessed. Moreover, it was possible to identify functional and RGC survival differences between groups by optomotor response (OMR), dark adapted full-field flash electroretinogram (ff-ERG), oscillatory potentials (OPs), photopic negative response (PhNR) and retinal flat-mount RGC counting 5 days after ONC. Comparisons between two groups were made using Mann-Whitney U test, unpaired t-test, unpaired t-test with Welch’s correction, one-way ANOVA, and Bonferroni t test. Results:Compared with the control group, the expression of HSPB8 protein in the retina of mice in ONC3 group was significantly increased, and the difference was statistically significant ( F=43.63, P<0.01). Compared with the control group, the ONC group showed obviously lower visual acuity ( P<0.01), lower a-wave, b-wave, OPs, PhNR amplitude, longer b-wave latency ( P<0.05), and the survival rates of RGC in ONC3 group, ONC5 group and ONC7 group decreased in a time-dependent manner( F=384.90, P<0.01). Transfection of AAV2 efficiency was highest on 4 weeks after IVT. Besides, there was no significant differences between the control group and the AAV2-shHspB8 group on visual acuity, ff-ERG, OPs, PhNR and RGC survival ( P>0.05). In comparison of the control group, we found that RGC survival of the ONC5+AAV2-shHspB8 group was significantly elevated ( F=10.62, P<0.01). Conclusions:Expression of HspB8 on the retina can be induced by ONC. The investigation of RGC counting, visual acuity, and ff-ERG revealed that optic nerve injury destructed functionality of mice retina and resulted to RGC death ultimately. The Most crucial finding of this research is that HspB8 knockdown had a neuroprotective effect in RGC after ONC.

Objective To improve the diagnosis and management of solid-pseudopapillary tumor (SPT) of the pancreas. MethodsTwenty-two SPT patients were retrospectively reviewed and divided into two groups, misdiagnosed group and those with preoperatively correct diagnosis. ResultsAbout one half (46%) SPT cases were misdiagnosed preoperatively. With time SPT tends to invade its capsule resulting in impossibility of radical resection, and increased medical expenses. ConclusionsAlthough SPT is of low degree malignant, and the prognosis after surgical resection is satisfactory, misdiagnosis and preoperative misdiagnosis and inappropriate management still cost the patients increased expenses and inhospital stay.

Objective To investigate the prognostic factors of differentiated thyroid carcinoma (DTC) in patients younger than 45 years. Methods Clinical data of all patients less than 45 years old at presentation with DTC in our hospital from Jan 1985 to Dec 1997 were analyzed retrospectively. Single variable analysis was performed by life-table method. Multivariate analysis was performed by Cox proportional hazard model. Results Two hundred and seventy two cases were analyzed. The overall 10-year survival rate was 93. 0%. The main prognostic factors influencing survival were age at presentation, the status of lymph node metastasis and distant metastasis; distant metastasis was the risk factor independently influencing survival by multivariate analysis. Conclusions Distant metastasis is the factor influencing survival significantly. The prognosis of these patients without distant metastasis is good. Total or near-total thyroidectomy and postoperative 131I therapy may be essential for a better prognosis in patients with distant metastasis.

Objective To explore the influence of MYC expression level on ovarian cancer cells adhesion and invasion ability,and the involvement of the integrinβ1 in the adhesion and invasion control. Methods Comparative study was done to analyze the relationship between the mRNA level of MYC and ITGB1 among sixty cell lines from the NCI-60 cells bank; Western blot was performed to demonstrate the correlation of MYC and ITGB1 protein level in ovarian cancer cell lines;Immunohistochemistry was adopted to compare the expression level of MYC and ITGB1 in paired primary and metastatic ovarian cancer tissue. Furthermore ,we employed in vitro adhesion test to detect the alteration of ovarian cancer cell adhesion ability to ECM molecular like fibronectin ,collagen I and laminin after fluctuation of MYC and ITGB1 expression; Transwell assay was applied to check the invasion ability variation with the disturbance of MYC and ITGB1 level. Results The mRNA levels of MYC and ITGB1 were dramatically inverse-correlated in NCI-60 cells bank(P<0.01);The protein levels of MYC and ITGB1 in ovarian cancer cell lines were also negative correlated; The primary ovarian cancer tissue showed lower expression of ITGB1 and higher expression of MYC,while the matched metastasis displayed the opposite outcome;Ovarian cancer cells reduced adhesive and invasive capacity in case of ITGB1 being down-regulated ,however, the adhesive and metastatic ability improved tremendously with elevation of ITGB1;Promoted adhesion and invasion ability and rise of ITGB1 expression were observed after downregulation of MYC,meanwhile,and inhibition of ITGB1 reversed the increasement of adhesion and invasion ability. Conclusions MYC,inversely-correlated with ITGB1 in ovarian cancer cells,inhibited ovarian cancer cells adhesion and invasion. Inhibition of MYC could lead to enhanced adhesion and invasion ability of ovarian cancer cells.

Objective:To investigate the effects of Krüppel-like factor 7 (KLF7) on the survival of retinal ganglion cells (RGCs) and electroretinogram (ERG) after retinal ischemia-reperfusion (RIR) injury in mice.Methods:A total of 126 male C57BL/6J mice were randomly divided into normal group, RIR group, normal-KLF7 group, normal-green fluorescent protein (GFP) group, RIR-KLF7 group and RIR-GFP group. At the age of 8 weeks, mice of normal-KLF7 group and RIR-KLF7 group were intravitreally injected 1ul of 1.0×10 12 vg/ml adeno-associated virus overexpressing KLF7 (AAV2-KLF7-GFP). Mice of normal-GFP group and RIR-GFP group were injected adeno-associated virus of AAV2-GFP with the same titer. At the age of 11 weeks, RIR injury was induced in mice of RIR group, RIR-KLF7 group and RIR-GFP group, and intraocular pressure was measured. Retinal cryosections were used to access the efficacy of virus transfection 4 weeks after AAV2-KLF7-GFP transfer. 7 days after RIR injury, RGCs' survival rate was observed and quantified by immunofluorescent staining. ERG was performed to observe the differences in amplitudes and incubation period of scotopic ERG a-, b-wave, oscillatory potentials (Ops), photopic negative responses (PhNR). Optomotor response was performed to observe the differences of visual acuity. Expression of KLF7 was detected by western blot 4 weeks after AAV2-KLF7-GFP transfer. Results:Compared with normal group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR group were decreased, and the differences were statistically significant ( t=12.860, 7.157, 5.735, 8.953, 4.744, 9.887; P<0.05). With the increase of light intensity, the amplitudes of scotopic ERG a- and b-wave were gradually increased while the incubation period was gradually shortened. Compared with RIR group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR-KLF7 group were increased, and the differences were statistically significant ( t=6.350, 3.253, 3.695, 5.825, 5.325, 4.591; P<0.05). Protein level of KLF7 was up-regulated in normal-KLF7 group than those in normal group, and the difference was statistically significant ( t=4.105, P<0.01). Conclusion:Overexpression of KLF7 can improve RGCs’ survival rates and preserve the electrophysiological function.

Objective To isolate endophytic fungi from Angelica sinensis and evaluate the bioactivity of their secondary metabolites.Methods Angelica sinensis and rhizosphere soil were utilized as materials.The tissue homogenization method was employed with six diverse culture media to isolate endophytic fungi.The antibacterial activity of secondary metabolites was gauged using a 96-well plate assay,while UV spectrophotometry was used to evaluate the inhibitory activity of four enzymes.Results A total of 153 fungal strains were isolated and purified from Angelica sinensis roots,stems,leaves,and soil.The samples exhibited specific inhibitory activities against adenosine deaminase(ADA),β-lactamase,xanthine oxidase(XO),and tyrosinase(TYR),with rates of 45.83%,52.78%,51.39%and 55.56%,respectively.Furthermore,1.39%of the samples displayed wide-ranging inhibitory effects against four indicator bacteria.Strain 6B also showcased the lowest inhibitory concentration values of 62.5 and 7.81 μg/mL against Escherichia coli ATCC25922 and ATCC35218,respectively,signifying its potential research significance.Conclusion Angelica sinensis has abundant endophytic fungal resources and is a good source for discovering active compounds,demonstrating certain research value.

OBJECTIVETo explore the effect of ataxia telangiectasia mutated and RAD3 related protein (ATR) expression and ATR kinase activity on the sensitivity to cisplatin in ovarian cancer SKOV3 cells.

METHODSSiRNA targeting ATR was transfected into SKOV3 cells for 48 h to reduce the ATR protein level, and ATR kinase inhibitor VE-821 was used for 12 h to inhibit the ATR pathway activity. The alteration of cell viability was examined by CCk-8 assay. Expression levels of ATR, p-ATR and γ-H2AX proteins were detected by Western blot. The DNA double strand breaks (DSB) marker γ-H2AX and homologous recombination repair key protein RAD51 and their co-localization in the cells were examined under the confocal microscope. The status of DNA double strand breaks (DSB) in single cells was visualized by alkaline comet assay. Finally, the cell cycle distribution was assessed using flow cytometry.

RESULTSDDP caused evident DNA double strands breaks and activated ATR kinase pathway. ATR-siRNA notably reduced ATR protein level, the 48 h IC(50) value of DDP was 72.12 µmol/L and 41.25 µmol/L, respectively, in the NC-siRNA and ATR-siRNA groups (P < 0.05). Confocal microscopic assay presented decreased recruitment of RAD51 at the DSB loci and comet assay showed enhanced DSB in the cells after ATR knocking down. After the inhibition of ATR kinase by VE-821, the 48 h IC(50) value of DDP was 75.32 µmol/L and 45.64 µmol/L, respectively, in the DMSO and VE-821 groups (P < 0.05 for both), confocal microscopic assay demonstrated reduced RAD51 recruitment, and comet assay showed increased DSB in cells after ATR kinase inhibition. Flow cytometry showed that percentage of cells distributed in G(0)/G(1), S and G(2)/M phases was 71.2%, 13.4% and 15.4%, repectively, after 40 µmol/L DDP treatment for 24 h. Compared with that of control group (G(0)/G(1): 54.2%, S: 21.3% and G(2)/M: 24.4%), DDP induced G(0)/G(1) phase arrest. DDP intervention resulted in the cell cycle status (G(0)/G(1): 43.2%, S: 20.4%, G(2)/M: 36.4%) in the ATR-siRNA group and (G(0)/G(1): 40.2%, S: 22.5%, G(2)/M: 37.3%) in the VE-821 group, indicating that the inhibition of ATR or ATR kinase could abrogate the effect of G(0)/G(1) phase arrest induced by DDP.

CONCLUSIONSSuppression of ATR can affect the homologous recombination repair in ovarian cancer cells, leading to accumulation of DNA double strand breaks in the cell nuclei as well as reduction of DDP-caused G(0)/G(1) phase arrest, finally enhances the sensitivity to cisplatin in the ovarian cancer SKOV3 cells.

Antineoplastic Agents ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; Cisplatin ; pharmacology ; DNA Repair ; Female ; Humans ; Ovarian Neoplasms ; Pyrazines ; RNA, Small Interfering ; Sulfones ; Transfection

Kidney deficiency syndrome is a common clinical syndrome in traditional Chinese medicine （TCM）. With the progress of science and technology， clinical and animal experiments on kidney deficiency syndrome have made remarkable progress. Research on kidney deficiency and the nature of "kidney" involves a large number of physiological and pathological bases， which are closely related to physiological and pathological links in the human body， among which the neuroendocrine-immune network shares the closest relationship. However， there are still many challenges in modern research on kidney deficiency syndrome， such as expert consensus on clinical diagnostic criteria and evaluation indexes and optimization of animal experimental models. In the past decade， a large number of clinical and animal experiments have been reported in the literature on kidney deficiency syndrome， among which the literature focusing on the combination of disease and syndrome is predominant， and most of them focus on kidney Yang deficiency and kidney Yin deficiency， involving the exploration of many pathological mechanisms. Research on the mechanisms related to kidney deficiency syndrome encompasses multiple signaling pathways and various biochemical indicators， including the phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-erythroid 2-relatedfactor-2（PI3K/Akt/Nrf2） signaling pathway， the Toll-like receptor 4/myeloid differentiation factor88/nuclear factor-κB（TLR4/MyD88/NF-κB） signaling pathway， the Janus kinase 2/signal transducer and activator of transcription 3（JAK2/STAT3） signaling pathway， Osteoprotectin/nuclear factor-κB receptor activator ligand/receptor activator of nuclear factor-κB （OPG/RANKL/RANK） signaling pathway. The biochemical indicators cover the cyclic adenosine monophosphate/cyclic guanosine monophosphate （cAMP/cGMP） ratio， Na⁺-K⁺-ATPase activity， Ca²⁺-Mg²⁺-ATPase activity， adrenocorticotropic hormone （ACTH）， polycorticosterone （CORT）， 17-OHCS， and other sex hormone indicators， providing crucial reference values for diagnosing kidney Yang deficiency or kidney Yin deficiency. The literature related to kidney deficiency syndrome over the past decade was collated and excavated， with a view to providing a reference for research on kidney deficiency syndrome.