[Therapeutic efficacy of transoral robotic surgery with the da Vinci robot system for the treatment of oropharyngeal squamous cell carcinoma].

Objective: To evaluate the clinical efficacy of transoral robotic surgery (TORS) with the da Vinci robot system in the treatment of oropharyngeal squamous cell carcinoma (OPSCC). Methods: A mixed cohort study was conducted to collect and analyze the clinical data of OPSCC patients who underwent TORS at the Eye & ENT Hospital, Fudan University between July 2020 and February 2023 (TORS group). OPSCC patients who underwent conventional surgery between January 2016 and September 2020 were included as the control group. The baseline information, incidence of complications and follow-up data were compared between the two groups. Results: A total of 166 patients were included, with 102 cases (81 males and 21 females) in the TORS group [mean age: (59.1±9.8) years] and 64 cases (54 males and 10 females) in the control group [ mean age: (57.6±9.7) years]. Compared with the control group, the TORS group had lower postoperative bleeding rate [2.9% (3/102) vs 10.9% (7/64), P=0.035] and infection rate [1.0% (1/102) vs 18.8% (12/64), P<0.001]. No statistically significant differences were observed in tracheotomy rate [46.1% (47/102) vs 59.4% (38/64), P=0.070] and median length of hospital stay [8 (7, 10) d vs 10 (4, 12) d, P=0.088]. After propensity score matching, compared with the control group, the TORS group had lower postoperative infection rate [0 (0/31) vs 19.4% (6/31), P=0.032] and median length of hospital stay [7 (7, 10) d vs 10 (8, 12) d, P=0.031]. No statistically significant differences were found in postoperative bleeding rate [3.2% (1/31) vs 6.5% (2/31), P=1.000] and tracheotomy rate [22.6% (7/31) vs 45.2% (14/31), P=0.060] between the two groups. Moreover, 1-and 2-year disease-free survival rates were 96.3% and 94.6% in the TORS group, and 90.6% and 84.3% in the control group, respectively (P=0.233). The 1-and 2-year cancer-specific survival rates were both 100% in the TORS group, and 96.9% and 93.8% in the control group, respectively (P=0.539). Conclusion: TORS for OPSCC is associated with high clinical safety and favorable oncological outcomes.

Predictive factors for enhanced community mental health vulnerability in this COVID-19 pandemic era.

OBJECTIVE: Explore the mental health status and its influencing factors of local community residents under the post-epidemic era of coronavirus disease 2019 (COVID-19) in China. METHODS: The basic information scale, self-rating depression scale and self-rating anxiety scale were used to carry out an online questionnaire survey among community residents in Jiangsu Province, China, and the influencing factors of depression and anxiety were analyzed by multivariate logistic regression. RESULTS: A total of 993 residents completed the mental health survey. It was found that the incidence of depressive and anxiety symptoms was 37.06% and 22.86%. Multivariate logistic regression analysis showed that women [odds ratio (OR) 95% confidence interval (95% CI) = 26.239 (14.743-46.698)], college degree and above [OR (95% CI) = 1.843 (1.085-3.130)] and ordinary residents [OR (95% CI) = 2.222 (1.441-3.425)] were risk factors for depressive symptoms, urban residents had lower risk [OR (95% CI) = 0.655 (0.394-0.829)]. Women [OR (95% CI) = 33.595 (15.812-71.381)] and ordinary residents [OR (95% CI) = 3.017 (1.602-5.680)] were risk factors for anxiety symptoms while the incidence was reduced in professional and technical personnel [OR (95% CI) = 0.271 (0.123-0.597)], workers [OR (95% CI) = 0.383 (0.168-0.876)], soldiers or policemen [OR (95% CI) = 0.200 (0.042-0.961)], married residents [OR (95% CI) = 0.463 (0.230-0.931)] and urban residents [OR (95% CI) = 0.531 (0.251-0.824)]. CONCLUSION: The incidence of symptoms of depression and anxiety among residents was relatively high under the post-epidemic era of COVID-19, which could be affected by various factors.

MiR-335-5p inhibits proliferation of Huh-7 liver cancer cells via targeting the Oct4/Akt pathway.

OBJECTIVE: The aim of this study was to detect the expression of micro ribonucleic acid (miR)-335-5p in the liver tissues of patients with liver cancer, and to explore its effect on liver cancer and mechanism using Huh7 human liver cancer cells. PATIENTS AND METHODS: Liver tissues were collected from patients with liver cancer. The expression of miR-335-5p in tissues was detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Subsequently, Huh7 cells were transfected with miR-335-5p in vitro. After overexpressing miR-335-5p, changes in the expression of octamer-binding transcription factor 4 (Oct4) gene were observed via qRT-PCR. Furthermore, the proliferation of Huh7 cells and the protein expressions of protein kinase B (Akt) and phosphorylated Akt (p-Akt) were detected using cell counting kit (CCK)-8 assay and Western blotting (WB), respectively. RESULTS: Compared with Control group, the expression of miR-335-5p increased significantly in the liver tissues of liver cancer patients (p<0.01). In comparison with those in negative group, the messenger RNA (mRNA) expression of Oct4 and the proliferation rate of Huh7 cells were both significantly inhibited in miR-335-5p group (p<0.01, p<0.05). After overexpression of miR-335-5p, the protein expression level of p-Akt decreased remarkably (p<0.01). CONCLUSIONS: MiR-335-5p directly binds to the 3' untranslated region (3'UTR) of Oct4 mRNA to restrain the phosphorylation of Akt, thereby inhibiting Huh7 cell proliferation.

[Value of fibrinogen to albumin ratio on predicting spontaneous recanalization of infarct-related artery in patients with acute ST-segment elevation myocardial infarction].

Objective: To investigate the value of fibrinogen to albumin ratio (FAR) at admission on predicting spontaneous recanalization of infarct-related artery (IRA) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods: Clinical data from 255 acute STEMI patients ((61.1±11.2) years old, 189 males) who underwent emergency coronary angiography within 12 hours in our hospital from December 2015 to April 2018 were retrospectively analyzed. The acute STEMI patients were divided into non-spontaneous recanalization group (thrombolysis in myocardial infarction (TIMI) flow grade 0-1, 203 cases) and spontaneous recanalization group (TIMI flow grade 2-3, 52 cases). Multivariate logistic regression analysis was used to evaluate related factors of IRA spontaneous recanalization. The receiver operating characteristic (ROC) curve was used to evaluate the value of FAR in predicting spontaneous coronary recanalization. Results: There was no significant difference in age,gender, hypertension, diabetes, smoking,systolic blood pressure,diastolic blood pressure,heart rate, duration of chest pain, type of infarction, infarct-related artery, door-to-balloon time, and drug used before admission between non-spontaneous recanalization group and spontaneous recanalization group (all P>0.05). The FAR and high-sensitivity C-reactive protein levels were significantly lower in the spontaneous recanalization group than in the non-spontaneous recanalization group (8.20±1.85 vs. 11.02±2.75, P<0.001; (6.87±3.36) g/L vs. (8.51±3.72) g/L, P=0.004). Multivariate logistic regression analysis showed that FAR (OR=0.492, 95%CI 0.354-0.686, P<0.001), serum uric acid (OR=0.994, 95%CI 0.989-0.999, P=0.018) and high-sensitivity C-reactive protein (OR=0.774, 95%CI 0.614-0.975, P=0.030) were independent negative correlation with spontaneous recanalization of infarct-related artery in patients with acute STEMI. The ROC curve showed that the area under the curve of FAR predicting spontaneous recanalization of infarct-related artery in patients with acute STEMI was 0.807 (95%CI 0.630-0.758, P<0.001), and the diagnostic threshold was 9.26, the sensitivity was 76.9%, the specificity was 75.9%. Conclusion: The level of admission FAR has certain predictive value for spontaneous recanalization of infarct-related arteries in patients with acute STEMI.

Two years efficiency of lamivudine and adefovir dipivoxil combined therapy in chronic hepatitis B patients.

BACKGROUND: Lamivudine (LAM) and adefovir (ADV) are widely used in most Asian countries, though monotherapy is associated with the occurrence of resistance. AIM: To evaluate the efficiency of LAM and ADV combined treatment of chronic hepatitis B patients with compensated cirrhosis. PATIENTS AND METHODS: 206 eligible Chinese patients were randomly assigned in a 1:1 ratio to receive either LAM or ADV for the first 24 weeks. According to virologic response at 24 weeks, the patients either continued to monotherapy or switched to combined therapy for 48 weeks. After 48 weeks, all patients received LAM and ADV combined therapy for 96 weeks. RESULTS: Serum HBV DNA levels significantly decreased in patients with ADV or LAM monotherapy and continuously reduced after the combined therapy. Serum ALT normalized rate were 88.24% and 81.37% at week 48, and 95.74% and 87.36% at week 96 in ADV and LAM group respectively, comparing to 60.78% and 56.73% in ADV and LAM groups at baseline. The accumulated virological breakthrough rate at week 48 and 96 was significantly higher in LAM group. CONCLUSIONS: Both combination strategies were resulted in the long term virological, biochemical improvement in Chinese chronic hepatitis B patients with compensated cirrhosis.

Roles of IL-4 and other factors in the trichosanthin-induced ovalbumin-specific IgE response.

AIM: To study the mechanism of trichosanthin (TCS)-induced ovalbumin (OVA)-specific immunoglobulin E (IgE) response in vivo. METHODS: To determine whether interleukin-4 (IL-4) was involved in TCS-induced IgE production, TCS and OVA co-immunized mice were treated with anti-IL-4 monoclonal antibodies (mAb) and OVA-specific serum IgE production was measured by enzyme-linked immunosorbent assay (ELISA). To distinguish whether recombinant IL-4 (rIL-4) was sufficient to support OVA-specific IgE response, OVA alone immunized mice were treated with rIL-4 and OVA-induced IgE production were examined by ELISA in the serum. To determine whether additional factors were involved in TCS-induced IgE response, th e kinetic expression of CD40 ligand (CD40L), tumor necrosis factor-alpha (TNF-alpha), and interleukin-13 (IL-13) were measured by semi-quantitative RT-PCR in draining mesenteric lymph node (MLN) from TCS-immunized mice. RESULTS: TCS-induced OVA-specific IgE production was suppressed by anti-IL-4 antibody, whereas IL-4 alone could not induce OVA-specific IgE production. CD40L, TNF-alpha, and IL-13 all expressed high levels in MLN after both primary and secondary immune responses. Among them CD40L had the similar transient expression peak to that of IL-4. CONCLUSION: IL-4 was indispensable for TCS-induced OVA-specific IgE production, and the other three factors examined may also be involved in, but CD40L may play a more important role.

Effect of valsartan and captopril in rabbit carotid injury. Possible involvement of bradykinin in the antiproliferative action of the renin-angiotensin blockade.

The effects of the specific angiotensin II (Ang II) AT1-receptor blocker valsartan on events related to restenosis were investigated in rabbits after common carotid balloon injury. Six animals were given valsartan from two days prior to injury until 14 days post-injury. Three control groups (n=6 in each group) were either sham-operated, untreated or treated with the angiotensin-converting enzyme (ACE) inhibitor,captopril. Both ACE inhibition and AT,-receptor blockade had marked effects on plasma levels of endothelin ET1, thromboxane TXB2 and 6-keto-PGF1-alpha. The most dramatic effects on ET, levels were seen in rabbits treated with valsartan, where levels were reduced to values close to those for sham-operated animals (96.85 vs. 86.45 pg/ml). Captopril treatment led to a statistically significant (p<0.01) reduction in ET1 levels compared with untreated animals, but the reduction was only about half that seen with AT1-receptor blockade. TXB2 levels doubled (202.58 vs.413.28 pg/ml) upon arterial injury in control animals but rose by only 20-35% in rabbits treated with captopril (246.45 pg/ml) or valsartan (268.13). In untreated animals, 6-keto-PGF1-alpha levels decreased slightly after injury, but for both the captopril and valsartan groups, there were significant increases in levels of this prostaglandin derivative, effects attributed to the action of bradykinins. Levels were highest in the captopril-treated animals. Valsartan and captopril treatment led to a significant reduction in neointimal thickness and the extent of lumen stenosis compared with untreated animals. Both treatments were effective in reducing neointimal area and significantly (p<0.05)reduced cell proliferation. The differences between treatments can be attributed to the different actions of the agents, as valsartan leaves the AT2-receptor unblocked, while captopril, through inhibition of Ang II synthesis, prevents stimulation of both receptors.A combination of both treatments may be a possible way forward in the clinical prevention of restenosis.

Long-term efficacy of recombinant interferon alpha 2a in the treatment of chronic hepatitis C: a randomized prospective study comparing two dose schedules in Chinese patients.

BACKGROUND/AIMS: This study is the first randomized prospective clinical trial of interferon in hepatitis to be conducted according to the guidelines of Good Clinical Practice (GCP) in China. The object of this study is to compare the long-term efficacy of a dose of 3MU of recombinant IFN alpha 2a (rIFN-alpha 2a) three times a week (t.i.w.) for 6 months with a starting dose of 6MU for 3 months and subsequent reduction to 3 MU t.i.w for a further 3 months. METHODOLOGY: Sixty-eight serological and histologically proven chronic hepatitis C patients with elevated serum ALT were randomized into two groups. A total of 63 patients were studied with full course of treatment. Five patients were withdrawn from the trial, 2 due to personal reasons and 3 due to adverse drug reactions during treatment. Thirty patients received 6MU IFN-alpha 2a t.i.w. for 3 months followed by 3MU t.i.w. for another 3 months (group A). Thirty-three patients received 3MU IFN-alpha 2a t.i.w. for 6 months (group B). RESULTS: The sex, age, baseline serum bilirubin, ALT and AST levels were matched in both groups. At the end of 6 months the complete and partial response rates in group A were 60.0% and 16.7%, respectively, and the clearance of serum HCV-RNA was 53.3%. In group B, the complete and partial response rates were 72.7% and 3.0%, respectively, and the clearance of HCV-RNA was 61.3%. These patients were followed up for 6, 12, and 18 months after stopping treatment. In group A, the rates of complete normalization of ALT and clearance of serum HCV-RNA at 24 months were 50.0% and 60.0%, respectively. In group B, the rates of normalization of ALT and clearance of HCV-RNA at 24 months were 54.4% and 41.9%, respectively. The efficacy between the two groups showed no statistically significant difference; the response rates of treatment were similar to the patients with HCV genotype 1b and 2a. Six patients (10.8% of the study population) developed neutralization antibodies to IFN-alpha 2a during treatment, 4 of them responded to the treatment. Adverse drug reactions (ADR) were common, but most of them were tolerable and the incidence of ADR was similar in both groups. CONCLUSIONS: IFN-alpha 2a is effective in the treatment of Chinese patients with chronic hepatitis C. The sustained response rates and ADR among two dose schedule groups are similar.

PSMA mimotope isolated from phage displayed peptide library can induce PSMA specific immune response.

Prostate-specific membrane antigen (PSMA) is a cell surface glycoprotein expressed predominantly in prostate secretory acinar epithelium and prostate cancer cells as well as in several extraprostatic tissues. Mouse monoclonal antibody 4G5 specific to the extracellular domain of PSMA was used to screen two phage displayed peptide libraries (9aa linear and 9aa cys library). Three 4G5-reactive phagotopes were identified. Sequence analysis of isolated clones demonstrated that the interaction motif "VDPA/SK" has high homology to 719-725aa on PSMA. Immunohistochemical staining of the prostate cancer sample with the PSMA-mimic phagotope (mimotope) immunized serum antibodies demonstrate that the mimotope isolated from the phage displayed peptide libraries can induce PSMA specific immune response in vivo.

The kinetics of IL-4 and IFN-gamma gene expression in mice after Trichosansin immunization.

Trichosanthin (TCS) is a potent allergen to mice. According to our previous experiments, it could bring out the IgE response to ovabumin (OVA) if TCS was given one day before OVA immunization, while OVA alone could not induce IgE to it. In this work, the kinetics of interleukin 4(IL-4) and interferon gamma(IFN-gamma) gene expression in the mesenteric lymph node (MLN) of TCS-immunized mice was investigated using a semi-quantitative RT-PCR method. It indicated that TCS induced significant IL-4 gene expression and the peaks of IL4 gene expression were on day one after TCS immunization in both primary and secondary response. In contrast, the IFN-gamma gene expression was suppressed. Furthermore, the IL-4 gene expression in the secondary response was lower than that in the primary response. Thus the presence of IgE memory B cells were studied. Results showed that the amount of mature IgE mRNA arose significantly and rapidly one day after TCS restimulation, while in the MLN of the mice primed 30 days before and without boost, it was almost as the same amount of the unimmunized control. These findings suggest the existence of the IgE memory B cells in the mice after the primary TCS immunization.

An improved rapid method for selecting monoclonal antibodies with high catalytic activities.

Monoclonal antibodies were raised against a beta-naphthyl phosphonate hapten (1) to elicit antibodies capable of catalyzing the hydrolysis of beta-naphthyl acetate (3). After cell fusion, potential catalytic antibody-producing hybridomas were selected, by use of a competitive inhibition assay on the basis of the binding activity for a short transition-state analogue (inhibitor 5), followed by use of high-performance liquid chromatography analysis for the hybridoma supernatants to screen the antibodies processing catalytic activities. It was shown that supernatants of 12 wells had high binding activity with inhibitor and of them, 7 had catalytic activities. After cloning by limiting dilution, we got two hybridoma clones producing monoclonal antibodies which catalyzed the hydrolysis of beta-naphthyl acetate. This combination of competitive inhibition assay with high-performance liquid chromatography analysis represents an improved rapid approach for the screening of potential catalytic antibodies and significantly increases the possibility of obtaining efficient catalytic monoclonal antibodies. Further study of the catalytic antibodies revealed significant rate enhancement (Kcat/K(uncat) approximately 10(6) and specificity.

Flow cytometry analysis of the neutralization effect of anti-IL-8 McAbs on IL-8-activated human granulocytes.

Two hybridoma clones, I8-S2 and I 8-63, secreting anti- IL-8 antibodies were constructed from fusion of immunized spleen cells of BALB/c mice with either Sp 2/0 or 653 myeloma cells. They recognized different epitopes on IL-8 molecule. IL-8 could activate human granulocytes and induce the elevation of [Ca2+]i. Through flow cytometry analysis the two clones displayed different neutralizing effects on the cell activation function of IL-8. The neutralizing effect of clone I8-S2 was apparently higher than that of clone I8-63. It is suggested that the cell activation function of IL-8 is restricted to a certain epitope of IL-8 molecule.

[Studies on immunohistochemical localization and immunotargeting of monoclonal antibody against human hepatocellular carcinoma].

Human hepatocellular carcinoma (HCC) cell line SMMC-7721 treated with human leucocytic alpha-interferon was used as immunogen to prepare monoclonal antibody against HCC. The anti-HCC monoclonal antibody (HAb23) was used to reacte with 51 cases of HCC, 38 cases of cirrhosis, 33 cases of hepatitis, 25 cases of other liver diseases, 36 cases of normal tissues including cryosections of fetus tissues and 231 cases of malignacies from various tissues by immunohistochemical staining. The results showed that 42 cases of HCC were positive, with a positive rate of 82.3%. Immuno-electron microscopic localization showed that HAb23 was localized mainly on the surface membrane and on the cytoplasmic membranous structures of tumor cells. The radioiodine conjugated HAb23 could target to the xenografts shown by ECT scanning. The ratios of tumor/liver and tumor/blood were 1.94-3.66 and 0.56-1.65, respectively. Five nm colloidal gold labeled HAb23 could also target to the xenografts. Our data suggest that the HAb23 is more specific to HCC and has a targeting ability to HCC in vivo.

[Immunohistochemical localization and serum testing for ferritin in malignant histiocytosis].

Localization of ferritin with immunohistochemical staining was carried out in thirty six cases of malignant histiocytosis (MH). The positivity rate for ferritin was 100 per cent. Ferritin was found to exist in the cytoplasm of the tumor cells. Image analysis showed that ferritin level in the well-differentiated histiocytes (1.2314) was higher than that in the atypical histiocytes (0.7181) (P < 0.01). Ten MH patients showed surprising high serum ferritin concentration (1482.3 ng/ml) than that in normal. Our data suggest that ferritin is the tumor associated antigen in MH. The synthesis and release of ferritin by MH tumor cells is the important cause for the high concentration of serum ferritin in patients.

[The establishment of human anti-tetanus toxoid hybridomas with in vitro immunized human tonsil cells].

Human tonsil cells in vitro immunized with tetanus toxoid were fused with human-mouse heteromyeloma line RF to generate human-mouse hybridomas. Hybridoma 891112-50 was cloned and 2 subclones (891112-50-3 and -4) were obtained. The secreted antibodies from the subclones were antigen specific, since they did not cross react with three irrelevant antigens (OVA, TCS and F gamma G). The hybridomas were quite stable. After 13 passages in tissue culture flasks, they still retained their antibody secreting ability. From flow cytometry analysis the subclone 50-3 was more stable than the subclone 50-4. The human immunoglobulin contained in supernatant collected during regular passages was equivalent to 69.6 micrograms/ml.

[Quantitative evaluation of the stability of antibody production by human-mouse hybridoma clones with flow cytometry analysis].

The paper described a method of quantitative stability analysis of antibody production by hybridoma clones with flow cytometry (FACS). Through FACS analysis the frequency of antibody-negative variants was measured. The rate of generation of antibody-negative variants/cell/generation (mu) could be calculated through Luria-Delbrück fluctuation analysis. The results showed that the value of mu was correlated to the antibody production of hybridoma clones: the larger was the mu value, the more unstable was the antibody production by the hybridoma clones. The experimental results suggested that the clone was stable if its mu value was on the magnitude of 10(-3), while the clone was not stable if its mu value was on the magnitude of 10(-2). According to requirements of single cell for FACS analysis and presence of a variety of methods for dispersing cell aggregates to single cells, the authors regard that this method is fully suitable for analysis of cells in suspension or even in the form of aggregate.

Radioimmunodetection and autoradiographic localization of monoclonal antibody against human hepatocellular carcinoma in xenografts.

The monoclonal antibody against human hepatocellular carcinoma, HAb23, was conjugated with radioiodine and injected into nude mice with human hepatoma xenografts. After 4 to 5 days, the nude mice were scanned by emission computerized topography. The results showed that the xenografts were distinguished clearly. The tumor-liver and tumor-blood ratios were 1.94 to 3.66 and 0.56 to 1.65, respectively. Autoradiography showed that the HAb23 was localized mainly on the surface membrane and tubulovesicular systems of the tumor cells.

[Cellular aspects of in vitro induction of antibody responses of human cells].

An in vitro system for induction of antibody responses of human cells has been established in our lab. B cell enriched fractions from excised human tonsils or trauma spleen were cultured for 7-14 days with tetanus toxoid or HBsAg vaccine with or without human T cell conditioned medium (C. M.) or a mixture of low concentrations of PWM and LPS (MTG). Positive antibody responses could be detected in cultures. Cells taken from different culture periods were subjected to FACS analysis in order to expound cellular changes during antibody induction periods so as to improve the in vitro antibody induction system. The results were described as follows: 1. Variations in total percentages of T cells during culturing periods seemed to be related its initial percentages. Cells with bigger initial percentages tended to decrease first and finally maintained at about 30%. While cells with smaller initial percentages tended to increase and finally also maintained at 30%. 2. CD4+ Th cells and CD8+ Ts cells from tonsils and spleen behaved somewhat differently. In tonsil cell cultures the percentages of CD4+ cells were often bigger than the percentages of CD8+ cells throughout the culture period. However, the inverted proportions of CD4+/CD8+ were shown in spleen cell cultures, especially in the culture with C. M. The possible relationships between the variations in CD4+/CD8+ proportions described as above and the intensities of antibody responses were discussed. Additionally, adding 1-Leucine-Methyl Ester showed no effects either on CD8+ or CD4+ cell percentages. 3. B cell (SIg+) percentages in both tonsil and spleen cultures were quite stable throughout the culture period, about 60% of total cells. CD19, a marker of B cell, was only present in part of the cultured SIg+ cells. The significance of the variations in CD19+, SIg+ cells was unclear. CD5+ B cells were known as cells secreting autoantibodies. Our results showed that these cells consistently maintained a relatively low percentage in the whole antibody induction period. 4. The reasonableness standard we used for "gating" in FACS analysis was discussed.