ObjectiveTo explore the molecular mechanisms by which serum containing Gegen Qinlian Tang （GQT） inhibits glycolysis and enhances chemotherapy sensitivity in 5-fluorouracil （5-FU）-resistant colorectal cancer （CRC） cells based on the cyclin-dependent kinase 16 （CDK16）/MYC proto-oncogene （MYC） pathway. MethodsHCT-116/5-FU cells were treated with different concentrations （5%， 10%， 20%， 30%） of GQT-containing serum. Cell viability and 5-FU sensitivity were assessed using the cell counting kit-8 （CCK-8） assay， and the experimental concentrations of 5-FU and GQT for subsequent experiments were determined. Cell proliferation and apoptosis under individual 5-FU， GQT， and combined 5-FU + GQT treatments were evaluated using 5-ethynyl-2′-deoxyuridine （EDU） staining and annexin V-FITC/PI double staining， respectively. Glucose consumption， adenosine triphosphate （ATP） production， and lactate levels were measured by colorimetric assays. Expression levels of glycolysis-related proteins， CDK16， MYC， and phosphorylated MYC were detected by Western blot. Co-immunoprecipitation （CoIP） was used to examine the protein interaction between CDK16 and MYC， and cycloheximide （CHX） treatment was applied to assess the effect of CDK16 overexpression on MYC protein stability. ResultsCCK-8 assays showed that 2.5 mg·L^-1 5-FU significantly inhibited HCT-116 cell viability in a dose-dependent manner. In HCT-116/5-FU cells， significant inhibition was observed only at 5 mg·L^-1 5-FU （P<0.05）， which was used for model establishment. Compared with 5-FU alone， addition of 5% GQT-containing serum significantly suppressed HCT-116/5-FU cell viability （P<0.05）， with stronger inhibition at higher serum concentrations. Thus， 5% GQT-containing serum was used in subsequent experiments. Compared with the control group， 5-FU， GQT， and 5-FU + GQT treatments all significantly reduced cell proliferation （P<0.05） and increased apoptosis （P<0.01）. The 5-FU + GQT combination showed superior inhibition of proliferation compared with 5-FU or GQT alone （P<0.01）， accompanied by more pronounced reductions in glucose consumption， ATP production， and lactate generation （P<0.01）. Additionally， compared with control， 5-FU， and GQT groups， the 5-FU + GQT group exhibited stronger suppression of MYC and its phosphorylated forms （P<0.01） and greater inhibition of glycolytic enzymes， including hexokinase 2 （HK2）， 3-phosphoinositide-dependent protein kinase 1 （PDK1）， lactate dehydrogenase A （LDHA）， and pyruvate kinase M2 （PKM2） （P<0.01）. CDK16， MYC， and MYC phosphorylation expression levels were significantly downregulated in the 5-FU + GQT group compared with the 5-FU group （all P<0.01）. MYC protein stability decreased in a time-dependent manner in the 5-FU + GQT group （P<0.05）， which was rescued by CDK16 overexpression （P<0.05）. ConclusionGQT significantly enhances the sensitivity of HCT-116/5-FU cells to 5-FU， potentially by inhibiting CDK16 and thereby reducing MYC-mediated glycolysis.

ObjectiveTo explore the molecular mechanisms by which serum containing Gegen Qinlian Tang （GQT） inhibits glycolysis and enhances chemotherapy sensitivity in 5-fluorouracil （5-FU）-resistant colorectal cancer （CRC） cells based on the cyclin-dependent kinase 16 （CDK16）/MYC proto-oncogene （MYC） pathway. MethodsHCT-116/5-FU cells were treated with different concentrations （5%， 10%， 20%， 30%） of GQT-containing serum. Cell viability and 5-FU sensitivity were assessed using the cell counting kit-8 （CCK-8） assay， and the experimental concentrations of 5-FU and GQT for subsequent experiments were determined. Cell proliferation and apoptosis under individual 5-FU， GQT， and combined 5-FU + GQT treatments were evaluated using 5-ethynyl-2′-deoxyuridine （EDU） staining and annexin V-FITC/PI double staining， respectively. Glucose consumption， adenosine triphosphate （ATP） production， and lactate levels were measured by colorimetric assays. Expression levels of glycolysis-related proteins， CDK16， MYC， and phosphorylated MYC were detected by Western blot. Co-immunoprecipitation （CoIP） was used to examine the protein interaction between CDK16 and MYC， and cycloheximide （CHX） treatment was applied to assess the effect of CDK16 overexpression on MYC protein stability. ResultsCCK-8 assays showed that 2.5 mg·L^-1 5-FU significantly inhibited HCT-116 cell viability in a dose-dependent manner. In HCT-116/5-FU cells， significant inhibition was observed only at 5 mg·L^-1 5-FU （P<0.05）， which was used for model establishment. Compared with 5-FU alone， addition of 5% GQT-containing serum significantly suppressed HCT-116/5-FU cell viability （P<0.05）， with stronger inhibition at higher serum concentrations. Thus， 5% GQT-containing serum was used in subsequent experiments. Compared with the control group， 5-FU， GQT， and 5-FU + GQT treatments all significantly reduced cell proliferation （P<0.05） and increased apoptosis （P<0.01）. The 5-FU + GQT combination showed superior inhibition of proliferation compared with 5-FU or GQT alone （P<0.01）， accompanied by more pronounced reductions in glucose consumption， ATP production， and lactate generation （P<0.01）. Additionally， compared with control， 5-FU， and GQT groups， the 5-FU + GQT group exhibited stronger suppression of MYC and its phosphorylated forms （P<0.01） and greater inhibition of glycolytic enzymes， including hexokinase 2 （HK2）， 3-phosphoinositide-dependent protein kinase 1 （PDK1）， lactate dehydrogenase A （LDHA）， and pyruvate kinase M2 （PKM2） （P<0.01）. CDK16， MYC， and MYC phosphorylation expression levels were significantly downregulated in the 5-FU + GQT group compared with the 5-FU group （all P<0.01）. MYC protein stability decreased in a time-dependent manner in the 5-FU + GQT group （P<0.05）， which was rescued by CDK16 overexpression （P<0.05）. ConclusionGQT significantly enhances the sensitivity of HCT-116/5-FU cells to 5-FU， potentially by inhibiting CDK16 and thereby reducing MYC-mediated glycolysis.

Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

Objective    To explore the association between radiotherapy and the risk of subsequent primary lung cancer in female patients with breast cancer. Methods    The EMbase, Web of Science, PubMed, CNKI, VIP and WanFang databases were searched from the establishment date to July 1, 2021. The relative risk (RR) with 95% confidence interval (CI) were combined and all statistical analyses were performed by STATA 12.0 software. Results    A total of 10 publications including 466 510 participates from 14 studies were included. The meta-analysis indicated that radiotherapy was a risk factor for the occurrence of second primary lung cancer among female breast cancer patients [RR=1.45, 95%CI (1.18, 1.78), P<0.001]. Subgroup analysis based on the relative position of breast cancer and lung cancer was conducted and the results demonstrated that radiotherapy only increased the incidence rate of ipsilateral lung cancer [RR=1.27, 95%CI (1.10, 1.45), P=0.001], without significant change of the risk of contralateral lung cancer [RR=1.16, 95%CI (0.77, 1.74), P=0.487]. Conclusion    Radiotherapy is one of the risk factors for subsequent primary pulmonary carcinoma among female breast cancer patients, especially for ipsilateral lung carcinoma. However, more high-quality studies are still needed to verify above findings.

Objective:To observe the effect of observing good swallowing on the swallowing action of stroke survivors with dysphagia.Methods:Eighteen stroke survivors with dysphagia were randomly divided into a treatment group ( n=9) and a control group ( n=9). In addition to routine swallowing rehabilitation therapy, the treatment group was asked to simulate swallowing after watching a video of normal people′s swallowing action. They did so 5 times a week for 10 minutes, while the control group just watched landscape videos at the same time. The treatment lasted 8 weeks. Before and after the treatment, both groups were assessed using the eating assessment tool (EAT-10), the functional oral intake scale (FOIS) and the penetration and aspiration scale (PAS). Functional magnetic resonance imaging (fMRI) was also used to observe their swallowing action. Results:There was no significant difference between the two groups in any of the measurements before the treatment. After the 8 weeks of treatment the average EAT-10, FOIS and PAS scores of the treatment group were all significantly better than before the treatment and better than the control group′s averages at the time. fMRI showed significantly more areas activated in the precuneus, parietal lobe, posterior central gyrus, BA7, BA5, frontal lobe and paracentral lobule in the treatment group compared with before the intervention and also more than in the control group.Conclusions:Observing proper swallowing action can improve dysphagia and activation of the swallowing-related brain areas of stroke survivors.

Objective:To observe the effect of applying repetitive transcranial magnetic stimulation (rTMS) on swallowing function and auditory evoked potentials in the brain-stems of stroke survivors with dysphagia.Methods:Sixty stroke survivors with dysphagia were randomly assigned to an observation group and a control group, each of 30. Both groups were given 30 minutes of routine swallowing training 6 times a week for 5 weeks, while the observation group was additionally provided with two-second bursts of rTMS on the left and right brain hemispheres alternately for 20 minutes daily. The frequency was 3.0Hz and the strength was 80% of the resting motor threshold (RMT), with an interval of 10 seconds between bursts. Before and after the treatment, both groups′ swallowing was assessed using the functional communication measure (FCM) and the modified Mann swallowing capacity evaluation scale (MMASA). Brainstem auditory evoked potentials (BAEP) were recorded to detect any changes in the peak latency (PL) and inter-peak latency (IPL). The clinical efficacy was also observed.Results:After the treatment, the average FCM results of both groups had improved significantly, but the observation group had improved significantly more. The average MMASA total scores and the average scores on all the sub-items had improved significantly more in the observation group except for the expressive speech disorder and anarthria scores. After the treatment, improvement was observed in respiration, anarthria, range of motion of the tongue muscles, strength of the tongue muscles, gag reflex and soft palate movement compared with before the intervention. Peak and inter-peak latencies had also improved significantly more in the observation than in the control group, on average.Conclusions:rTMS combined with conventional swallowing training can significantly improve the swallowing of dysphagic stroke survivors, and shorten the PL of the BAEP so as to better regulate deglutition.

Objective 聽 聽To investigate the correlation between the postoperative day 1 (POD1) drainage volume and postoperative pulmonary complications (PPCs). Methods 聽 聽The clinical data of 254 non-small cell lung cancer (NSCLC) patients undergoing thoracoscopic lobectomy at our department from January 2014 to June 2018 were retrospectively reviewed. According to whether there were PPCs after operation, patients were divided into a PPCs group (51 patients, 33 males and 18 females, aged 65.2卤7.3 years) and a non-PPCs group (203 patients, 110 males and 93 females, aged 62.4卤8.2 years). The correlation between POD1 drainage volume and PPCs was analyzed. Results 聽 聽The POD1 drainage volume in the PPCs group was significantly more than that in the non-PPCs group (337.5卤127.4 ml vs. 208.7卤122.9 ml, P=0.000). The result of regression analysis showed that POD1 drainage volume was an independent risk factor for the occurrence of PPCs. Receiver operating characteristic curve (ROC curve) analysis showed that POD1 drainage volume of 265 ml was the cut-off point to predict PPCs. The rate of PPCs in the group of POD1 drainage volume less than 265 ml was significantly lower than that in the group of drainage volume more than 265 ml (8.3% vs. 43.0%, P=0.000). Conclusion 聽 聽The POD1 drainage volume is closely related to the occurrence of PPCs, which can be used to predict the occurrence of PPCs.

Objective To investigate the safety of training using a lower-limb rehabilitation robot and its effectiveness in terms of stroke survivors' cardiopulmonary function.Methods Thirty-two hemiplegic stroke survivors were randomly divided into a robot group and a control group,each of 16.Both groups received conventional rehabilitation medication and training,120 min/d,5 d/week for 8 weeks.The robot group was additionally trained with a Flexbot robotic gait training apparatus for 30 min/d,5 d/week for the same 8 weeks.The subjects' maximum oxygen consumption (VO2max),heart rate,blood pressure,ventilation (VE) and rate of perceived exertion (RPE) were quantified before and after the training.Results After the 8 weeks there was no significant difference in average heart rate,blood pressure,VE or RPE between the two groups.The average VO2max of the robot group was,however,significantly higher than that of the control group.Conclusion Robotic gait training is safe and can improve the cardiopulmonary function of stroke survivors.

Objective To observe the effect and safety of the endovascular hypothermia through hypothermic intravenous infusion device in the treatment of patients with severe craniocerebral injury compared with conventional surface cooling. Methods A total of 66 cases of patients with severe cranio-cerebral injury were randomly divided into the observation group and the control group with 33 cases in each group according to envelop randomization. The control group received surface cooling, and the observation group was given surface cooling plus endovascular hypothermia through hypothermic intravenous infusion device. The target temperature was 35 ℃maintained for 3-5 days, and natural rewarming was applied at the speed of 0.1-0.5℃/h to 36.0-37.3℃. The time to reach target temperature, the constant stability, the incidence rate and severity of complication such as shiver, arrhythmia, skin injury and agitation were recorded and compared between two groups, as well as the heart rate, breathing rate, pulse rate, blood pressure and Glasgow Coma Scale (GCS) scores after 72 h of treatment. Glasgow Outcome Scale scores 30 days after treatment and nursing workload were also calculated and compared. Results The cooling speed, time to reach target temperature and the ability to maintain at 35℃were (1.3±0.2)℃/h, (2.3±0.2) h and (6.5± 1.8)%in the observation group, respectively, compared with (0.5±0.1)℃/h, (3.6±0.6) h and (11.3±2.2)%in the control group, which had significant differences (t=1.862, 2.112, 2.408,P < 0.05). The occurrence rates of shiver, arrhythmia, skin damage and dysphoria and restlessness in the observation group were 33.33%(11/33), 9.09%(3/33), 6.06%(2/33) and 27.27%(9/33), respectively, which were much lower than those in the control group 84.85%(28/33), 15.15%(5/33), 33.33%(11/33), 54.55%(18/33),χ2=1.764-2.733,P<0.05. The heart rate, breathing rate, pulse rate, systolic blood pressure and GCS score after 72 h of treatment were(68.31 ± 3.73)times/min,(16.60 ± 1.52)times/min,(136.35 ± 3.71)mmHg(1 mmHg=0.133 kPa),(34.61 ± 1.05)℃, (9.91±4.05)points in the observation group, while(58.31±3.62)times/min,(19.81±1.83)times/min,(150.66± 2.70)mmHg,(35.65 ± 1.36)℃,(7.63 ± 3.17)points in the control group, and there were significant differences between two groups(t=2.275-3.035, P < 0.05).Besides, the ice-changing ice and turning-over time in the observation group were both remarkably reduced compared with control group, (14.03±3.11) min/h vs (38.12± 2.70) min/h (t=3.356, P<0.05) , (15.08±3.07) min/h vs (26.16±2.54) min/h ( t=3.021, P<0.05). Patients with good recovery, mild disability, severe disability, death in the observation group were 16, 13, 3 and 1 case, while 6, 11, 9, 7 cases in the control group (χ2=2.351,P < 0.05). Conclusions The endovascular hypothermia through hypothermic intravenous infusion device can rapidly reduce and effectively maintain target temperature, reduce the incidence rate of complication, improve the vital signs and decrease the nursing workload in order to improve neurological outcome in the treatment of patients with severe craniocerebral injury.

Objective To investigate the application effect of stone basket in ureteroscopic holmium laser lithotripsy for treatment of upper ureteral calculi. Methods The clinical data of 96 patients with upper ureteral calculi were retrospectively analyzed, all patients underwent ureteroscopic holmium laser lithotripsy. Forty-eight cases used the stone basket in the operation process (observation group), and 48 cases did not use the stone basket in the operation process (control group). The operation time, length of stay, success rate of lithotripsy, stone residual rate and incidence of postoperative complication were compared between 2 groups. Results The patients of 2 groups successfully completed surgery. There were no statistical differences in operation time, length of stay and incidence of postoperative complication between 2 groups (P>0.05). The success rate of lithotripsy in observation group was significantly higher than that in control group: 97.92% (47/48) vs. 75.00% (36/48), and the stone residual rate was significantly lower than that in control group:4.17%(2/48) vs. 18.75%(9/48), and there were statistical differences (P<0.05). Conclusions The stone basket in ureteroscopic holmium laser lithotripsy for treatment of upper ureteral calculi can thoroughly remove stones. It reduces the incidence of residual stones, does not affect the safety of the operation, and has good clinical value.