The efficient clinical treatment of oral squamous cell carcinoma(OSCC)is still a challenge that demands the development of effective new drugs.Phenformin has been shown to produce more potent anti-tumor activities than metformin on different tumors,however,not much is known about the influence of phenformin on OSCC cells.We found that phenformin suppresses OSCC cell proliferation,and promotes OSCC cell autophagy and apoptosis to significantly inhibit OSCC cell growth both in vivo and in vitro.RNA-seq analysis revealed that autophagy pathways were the main targets of phenformin and identified two new targets DDIT4(DNA damage inducible transcript 4)and NIBAN1(niban apoptosis regulator 1).We found that phenformin significantly induces the expression of both DDIT4 and NIBAN1 to promote OSCC autophagy.Further,the enhanced expression of DDIT4 and NIBAN1 elicited by phenformin was not blocked by the knockdown of AMPK but was suppressed by the knockdown of transcription factor ATF4(activation transcription factor 4),which was induced by phenformin treatment in OSCC cells.Mechanistically,these results revealed that phenformin triggers endoplasmic reticulum(ER)stress to activate PERK(protein kinase R-like ER kinase),which phosphorylates the transitional initial factor eIF2,and the increased phosphorylation of eIF2 leads to the increased translation of ATF4.In summary,we discovered that phenformin induces its new targets DDIT4 and especially NIBAN1 to promote autophagic and apoptotic cell death to suppress OSCC cell growth.Our study supports the potential clinical utility of phenformin for OSCC treatment in the future.

Objective:To observe the clinical efficacy of mouse nerve growth factor (mNGF) combined with rehabilitation on children with global developmental delay(GDD).Methods:It was a prospective multicenter clinical randomized controlled trial (RCT) involving 120 children with GDD admitted to 5 hospitals in China from May 2020 to January 2022.They were randomly divided into mNGF group and conventional rehabilitation group using block randomization method.All children were managed by standardized rehabilitation after recruitment, and those in the mNGF group were additionally given mNGF injections.All subjects were surveyed using the Gesell Development Diagnosis Schedules(GDDS) at baseline, 90 days and 120 days after treatment, and their developmental quotient (DQ) was recorded.Clinical efficacy was analyzed by the paired t-test, rank sum test and Chi- squared test. Results:After 90 days of treatment and the continuous follow-up to 120 days, the increases in the DQ of gross motor (7.520±13.900 vs.0.450±11.459), fine motor (7.800±15.346 vs.1.250±11.581), adaptive behavior (7.730±13.428 vs.2.100±12.022) and personal-social behavior (6.780±11.651 vs.1.780±10.120) than baseline were significantly higher in mNGF group than those of conventional rehabilitation group (all P<0.05). Serious adverse events and important drug-related medical events were not reported. Conclusions:mNGF combined with rehabilitation effectively enhances the development levels of gross motor, fine motor, adaptive behavior and personal-social behavior, and continuously improves the condition of GDD in children with a high safety.

The aim of this study was to construct a recombinant adenovirus expressing extracellular domain gene of human epidermal growth factor receptor variant Ⅲ (EGFRvIII ECD), and to prepare single domain antibody targeting EGFRvIII ECD by immunizing camels and constructing phage display antibody library. Total RNA was extracted from human prostate cancer cell line PC-3 cells and reversely transcribed into cDNA. EGFRvIII ECD gene was amplified using cDNA as template, and ligated into pAdTrack-CMV plasmid vector and transformed into E. coli BJ5183 competent cells containing pAdEasy-1 plasmid for homologous recombination. The recombinant adenovirus expressing EGFRvIII ECD was obtained through transfecting the plasmid into HEK293A cells. The recombinant adenovirus was used to immunize Bactrian camel to construct EGFRvIII ECD specific single domain antibody library. The single domain antibody was obtained by screening the library with EGFRvIII protein and the antibody was expressed, purified and identified. The results showed that recombinant adenovirus expressing EGFRvIII ECD was obtained. The capacity of EGFRvIII specific phage single domain antibody library was 1.4×10⁹. After three rounds of enrichment and screening, thirty-one positive clones binding to EGFRvIII ECD were obtained by phage-ELISA, and the recombinant single domain antibody E14 with highest OD₄₅₀ value was expressed and purified. The recombinant E14 antibody can react with EGFRvIII ECD with high affinity in ELISA assessment. The results indicated that the EGFRvIII specific single domain antibody library with high capacity and diversity was constructed and the single domain antibody with binding activity to EGFRvIII was obtained by screening the library. This study may facilitate the diagnosis and treatment of EGFRvIII targeted malignant tumors in the future.
Adenoviridae/genetics* ; DNA, Complementary ; ErbB Receptors ; Escherichia coli/genetics* ; Genetic Vectors/genetics* ; Humans ; RNA ; Recombinant Proteins/metabolism* ; Single-Domain Antibodies

Objective: To investigate the relationship between systemic inflammatory markers such as neutrophil/lymphocyte ratio (NLR) and C-reactive protein/albumin ratio (CAR), and lymph node metastasis in patients with cN0 gastric cancer. To establish a nomogram model to predict the risk of lymph node metastasis in patients with cN0 gastric cancer. Methods: The preoperative systemic inflammatory markers and clinical data of 134 patients with cN0 gastric cancer were retrospectively analyzed, and these markers of patients with negative (pN0) or positive (pN+ ) lymph node metastasis in postoperative pathological diagnosis were compared. The receiver operating characteristic (ROC) curve was used to evaluate the predictive effect of preoperative systemic inflammatory markers on lymph node metastasis. The influencing factors for lymph node metastasis were assessed by univariate analysis and multivariate logistic regression analysis. A nomogram subsequently established by R software was validated by Bootstrap resampling as internal validation. Results: Compared with pN0 group, NE (P=0.022), CRP (P<0.001), NLR (P<0.001), PLR (P=0.003) and CAR (P<0.001) were higher, LY (P=0.003) and Alb (P=0.042) were lower in pN+ group. ROC curve analysis showed that the area under the curve (AUC) of postoperative pathological lymph node metastasis in patients with cN0 gastric cancer diagnosed by NLR, PLR and CAR were 0.687, 0.651 and 0.694, respectively, and the best cutoff values were 2.12, 113.59 and 0.02, respectively. The corresponding sensitivity and specificity were 62.9% and 72.2%, 77.4% and 48.6%, 74.2% and 58.3%, respectively. Univariate analysis showed that tumor size, depth of invasion, NLR, PLR and CAR were associated with lymph node metastasis in cN0 gastric cancer patients (all P<0.05). Multivariate analysis showed that depth of invasion, NLR and CAR were independent influencing factors of lymph node metastasis in patients with cN0 gastric cancer. OR were 8.084, 3.540 and 3.092, respectively (all P<0.05). The C-index of the nomogram model was 0.847 (95% CI: 0.782-0.915). The predicting calibration curve was properly fit with the ideal curve in calibration chart. Conclusion Combination of NLR and CAR to establish a nomogram model has a good consistency and can accurately predict the risk of lymph node metastasis in patients with cN0 gastric cancer.

Weaning?failure?from?mechanical?ventilation?is?an?important?clinical?problem,?the?traditional?methods?of?assessing?whether?patients?can?be?weaned?from?mechanical?ventilation?or?not?cannot?meet?the?clinical?needs.?Finding?more?effective?weaning?indicators?to?determine?the?optimal?timing?of?weaning?has?important?clinical?value?for?improving?the?outcome?of?weaning?and?reducing?the?mortality?of?patients.?As?a?new?method?of?assessing?respiratory?function?of?patients,?electrical?impedance?tomography?(EIT)?is?gradually?applied?to?the?clinic,?and?its?guided?assessment?of?respiratory?function?may?open?a?new?way?for?directing?successful?weaning.?This?article?reviews?the?progress?of?EIT?in?ventilation?weaning,?in?order?to?provide?a?new?judgment?method?and?theoretical?basis?for?the?successful?weaning.

Objective To investigate the relationship between systemic inflammatory markers such as neutrophil／lymphocyte ratio ( NLR) and C?reactive protein／albumin ratio ( CAR ), and lymph node metastasis in patients with cN0 gastric cancer. To establish a nomogram model to predict the risk of lymph node metastasis in patients with cN0 gastric cancer. Methods The preoperative systemic inflammatory markers and clinical data of 134 patients with cN0 gastric cancer were retrospectively analyzed, and these markers of patients with negative ( pN0 ) or positive ( pN+) lymph node metastasis in postoperative pathological diagnosis were compared. The receiver operating characteristic ( ROC ) curve was used to evaluate the predictive effect of preoperative systemic inflammatory markers on lymph node metastasis. The influencing factors for lymph node metastasis were assessed by univariate analysis and multivariate logistic regression analysis. A nomogram subsequently established by R software was validated by Bootstrap resampling as internal validation. Results Compared with pN0 group, NE (P＝0.022), CRP (P＜0.001), NLR (P＜0.001), PLR (P＝0.003) and CAR ( P＜0.001) were higher, LY ( P＝0.003) and Alb ( P＝0.042) were lower in pN+ group. ROC curve analysis showed that the area under the curve ( AUC) of postoperative pathological lymph node metastasis in patients with cN0 gastric cancer diagnosed by NLR, PLR and CAR were 0.687, 0.651 and 0.694, respectively, and the best cutoff values were 2.12, 113.59 and 0.02, respectively. The corresponding sensitivity and specificity were 62.9% and 72.2%, 77.4% and 48.6%, 74.2% and 58.3%, respectively. Univariate analysis showed that tumor size, depth of invasion, NLR, PLR and CAR were associated with lymph node metastasis in cN0 gastric cancer patients ( all P＜0.05). Multivariate analysis showed that depth of invasion, NLR and CAR were independent influencing factors of lymph node metastasis in patients with cN0 gastric cancer. OR were 8.084, 3.540 and 3.092, respectively ( all P＜0.05). The C?index of the nomogram model was 0.847 (95% CI: 0.782?0.915). The predicting calibration curve was properly fit with the ideal curve in calibration chart.Conclusion Combination of NLR and CAR to establish a nomogram model has a good consistency and can accurately predict the risk of lymph node metastasis in patients with cN0 gastric cancer.

Objective To investigate the relationship between systemic inflammatory markers such as neutrophil／lymphocyte ratio ( NLR) and C?reactive protein／albumin ratio ( CAR ), and lymph node metastasis in patients with cN0 gastric cancer. To establish a nomogram model to predict the risk of lymph node metastasis in patients with cN0 gastric cancer. Methods The preoperative systemic inflammatory markers and clinical data of 134 patients with cN0 gastric cancer were retrospectively analyzed, and these markers of patients with negative ( pN0 ) or positive ( pN+) lymph node metastasis in postoperative pathological diagnosis were compared. The receiver operating characteristic ( ROC ) curve was used to evaluate the predictive effect of preoperative systemic inflammatory markers on lymph node metastasis. The influencing factors for lymph node metastasis were assessed by univariate analysis and multivariate logistic regression analysis. A nomogram subsequently established by R software was validated by Bootstrap resampling as internal validation. Results Compared with pN0 group, NE (P＝0.022), CRP (P＜0.001), NLR (P＜0.001), PLR (P＝0.003) and CAR ( P＜0.001) were higher, LY ( P＝0.003) and Alb ( P＝0.042) were lower in pN+ group. ROC curve analysis showed that the area under the curve ( AUC) of postoperative pathological lymph node metastasis in patients with cN0 gastric cancer diagnosed by NLR, PLR and CAR were 0.687, 0.651 and 0.694, respectively, and the best cutoff values were 2.12, 113.59 and 0.02, respectively. The corresponding sensitivity and specificity were 62.9% and 72.2%, 77.4% and 48.6%, 74.2% and 58.3%, respectively. Univariate analysis showed that tumor size, depth of invasion, NLR, PLR and CAR were associated with lymph node metastasis in cN0 gastric cancer patients ( all P＜0.05). Multivariate analysis showed that depth of invasion, NLR and CAR were independent influencing factors of lymph node metastasis in patients with cN0 gastric cancer. OR were 8.084, 3.540 and 3.092, respectively ( all P＜0.05). The C?index of the nomogram model was 0.847 (95% CI: 0.782?0.915). The predicting calibration curve was properly fit with the ideal curve in calibration chart.Conclusion Combination of NLR and CAR to establish a nomogram model has a good consistency and can accurately predict the risk of lymph node metastasis in patients with cN0 gastric cancer.

Objective To study the influence of semen platycladi saponins on oxidative stress response of hippocampus of rat model of Alzheimer’s disease (AD), and to explore the neuroprotective mechanism of semen platycladi saponins (SPS) on AD model rats. Methods Sixty SPF 24-week old female Wistar rats were divided into three groups:the normal control group, AD model group, and SPS intervention group. The rats of AD model group and SPS group were injected with Aβ1-42in bilateral hippocampi to produce AD animal models. After the successful establishment of AD model, the AD model group and the normal control group received 5 mL saline sodium with carboxymethyl cellulose (500 mg/kg) orally, daily for 30 days. The SPS group received orally 5 mL normal saline with 300 mg/kg SPS daily for 30 days. The learning and memory function of the rats were assessed by Morris water maze test, and the levels of MAD, SOD and GSH in the hippocampal tissues of model rats were detected with biochemistry. The expressions of Bcl-2, survivin, Fas, Bax, caspase-3 proteins and mRNA in the hippocampi of AD model rats were detected by western blotting. Results Compared with the control group and SPS group, the rats in AD model group displayed a longer search time and shorter percentage of search distance (P＜ 0. 01). There was a longer search time and lower percentage of search distance of the SPS groups than the control group (P＜ 0. 01). The positioning experiment showed that rats in the control group learned to find the platform within 2. 1 days, indicating that the latency was rapidly decreased. MDA in the SPS intervention group was significantly decreased than in the AD model group (P＜ 0. 01), but increased than the normal control group (P＜0. 01). The expressions of SOD and GSH were significantly increased in the SPS group than the model group (P＜ 0. 01). The expressions of Bcl-2 and survivin were significantly increased in the SPS group than the model group (P＜ 0. 01), but lower than the normal control group (P＜ 0. 05). The expressions of Fas, Bax and caspase-3 were decreased in the SPS intervention group (P ＜ 0. 01 ), but increased than the normal control group (P ＜ 0. 05 ). Conclusions Semen platycladi saponins can protect the neurons and improve the cognition function of AD model rats by inhibiting oxidative stress response and enhancing the antioxidant mechanism in the hypocampus.

Rectal cancer is one of the most common malignant tumor in our country,among them with highest incidence of low rectal cancer.With the further study on the biological behavior of rectal cancer and the development of diagnostic and therapeutic techniques,gastrointestinal oncologists have proposed a new treatment strategy-local resection and wait-and-see.This strategy can better preserve anal function and improve the quality of life of patients without losing the good tumor control of the traditional standard treatment model.

Objective To divide computed tomography (CT) values into different ranges and investigate the influence of CT value division on dose calculation, and to propose a method to combine magnetic resonance imaging (MRI) with assigned CT values.Methods Ten CT images each were collected from patients with head and neck, chest, and pelvic tumors.Random sampling was performed for the CT values of main tissues or organs at the three parts, and then the mean CT value of each tissue or organ was calculated to divide the CT values into different ranges.A virtual phantom was built in the Varian Eclipse treatment planning system, and for the prescribed dose of 100 cGy, the machine output was recorded at different CT values.The influence of different CT value ranges on dose calculation was analyzed.The treatment plans of intensity-modulated radiotherapy were selected from 5 cervical cancer patients, and new CT values were assigned to the planning target volume (PTV) and organs at risk to obtain new CT images.The plans were transferred to the new CT images and compared with the results on the original CT images in terms of dosimetric parameters.Results After dividing the CT values into different ranges and verifying the results in dose calculation, the CT values corresponding to different human tissues or organs were-100 to 100 HU.The influence of CT value variation on dose calculation was within 3%.In the same treatment plan, there were small differences in dosimetric parameters between new CT images and original CT images.Dmax, Dmean, D98%, D95%, D5%, and D2% of PTV were all below 3%, and Dmax and Dmean of the bladder, rectum, small intestine, femoral head, and bone marrow were below 2%.Conclusions The influence of CT value division on dose calculation in the treatment planning for pelvic tumors is acceptable, so it can be used in combination with MRI.