Guided by the turbidity toxin theory， it is believed that the key pathogenesis of constipation-predominant irritable bowel syndrome is the obstruction of turbidity toxin and the disruption of intestinal function. Treatment is based on the principles of dispelling turbidity toxin and promoting intestinal function. The clinical patterns can be divided into three types， turbidity toxin heat accumulation pattern， turbidity toxin combined with liver depression and qi stagnation pattern， and turbidity toxin combined with qi and yin deficiency pattern. The treatment can respectively use self-prescribed Tongfu Jiangzhuo Formula （通腑降浊方） to clear heat and unblock the bowels， direct the turbid downward and resolve toxins； use self-prescribed Shugan Jiangzhuo Formula （疏肝降浊方） to soothe the liver and move qi， direct the turbid downward and resolve toxins； use self-prescribed Mazhi Jiangzhuo Formula （麻枳降浊方） to boost qi and nourish yin， moisten the intestines to remove turbidity and resolve toxins.

Objective : To investigate the brain microvascular tissue block culture method for extracting primary rat brain microvascular endothelial cells and identify its effect. Methods : Brain tissue from 4-week-old Sprague Dawley rats was screened, pre-digested and solidified to obtain brain microvascular segments. These segments were subsequently placed in a CO2incubator for primary culture. The target cells were identified by cell morphology and immunocytochemical staining for factor Ⅷ-related antigen. Results : After a 48-hour culture periodin vitro, the short spindle cells crawled out from around the brain microvascular segments. After 72 hours, island-like cell culsters formed. After 96 hours the clusters fused and the cells formed a typical monolayer, cobble stone-like, and mosaic arrangement. Factor Ⅷ-related antigen immunocytochemical staining showed that the cytoplasm of the cells appeared brown-red, indicating positive expression; DAB stained the nucleus, showing blue-dark. Conclusion The brain microvascular tissue block culture method can isolate and culture primary rat brain microvascular endothelial cells.

BACKGROUND: The purpose of this study was to evaluate the safety and efficacy of subsequent radiotherapy (RT) following first-line treatment with durvalumab plus chemotherapy in patients with extensive-stage small cell lung cancer (ES-SCLC). METHODS: A total of 122 patients with ES-SCLC from three hospitals during July 2019 to December 2021 were retrospectively analyzed. Inverse probability of treatment weighting (IPTW) analysis was performed to address potential confounding factors. The primary focus of our evaluation was to assess the impact of RT on progression-free survival (PFS) and overall survival (OS). RESULTS: After IPTW analysis, 49 patients received durvalumab plus platinum-etoposide (EP) chemotherapy followed by RT (Durva + EP + RT) and 72 patients received immunochemotherapy (Durva + EP). The median OS was 17.2 months vs . 12.3 months (hazard ratio [HR]: 0.38, 95% confidence interval [CI]: 0.17-0.85, P = 0.020), and the median PFS was 8.9 months vs . 5.9 months (HR: 0.56, 95% CI: 0.32-0.97, P = 0.030) in Durva + EP + RT and Durva + EP groups, respectively. Thoracic radiation therapy (TRT) resulted in longer OS (17.2 months vs . 14.7 months) and PFS (9.1 months vs . 7.2 months) compared to RT directed to other metastatic sites. Among patients with oligo-metastasis, RT also showed significant benefits, with a median OS of 17.4 months vs . 13.7 months and median PFS of 9.8 months vs . 5.9 months compared to no RT. Continuous durvalumab treatment beyond progression (TBP) prolonged OS compared to patients without TBP, in both the Durva + EP + RT (NA vs . 15.8 months, HR: 0.48, 95% CI: 0.14-1.63, P = 0.238) and Durva + EP groups (12.3 months vs . 4.3 months, HR: 0.29, 95% CI: 0.10-0.81, P = 0.018). Grade 3 or 4 adverse events occurred in 13 (26.5%) and 13 (18.1%) patients, respectively, in the two groups; pneumonitis was mostly low-grade. CONCLUSION Addition of RT after first-line immunochemotherapy significantly improved survival outcomes with manageable toxicity in ES-SCLC.
Humans ; Small Cell Lung Carcinoma/therapy* ; Retrospective Studies ; Male ; Female ; Middle Aged ; Lung Neoplasms/therapy* ; Aged ; Antibodies, Monoclonal/therapeutic use* ; Adult ; Immunotherapy/methods* ; Aged, 80 and over

The Chinese hamster ovary (CHO) cell is the most representative mammalian cell protein expression system, and it is widely used in recombinant protein, vaccine and other biopharmaceutical fields. However, due to its vulnerability to environmental factors, apoptosis, and metabolic inhibitors, CHO cells demonstrate poor robustness, and thus the integrated viable cell density and unit cell productivity are largely limited. To improve the robustness and foreign protein expression efficiency of CHO cells, we employed CRISPR/Cas9 to knock out the apoptosis genes Bax and Bak and the lactate dehydrogenase gene LDHa, thereby blocking apoptosis and lactic acid metabolism pathways. The results of apoptosis and single cell viability detection showed that the number of apoptotic cells in the knockout cell lines Bax^-/-, Bax-bak^-/-, and LDHa-Bax-bak^-/- was reduced by 22.51%, 37.73%, and 64.12%, respectively, compared with the wild-type cell line CHO-K1, which indicated that the anti-apoptotic ability was significantly improved. After staurosporine treatment, the single cell viability of Bax^-/-, Bax-bak^-/-, and LDHa-Bax-bak^-/- cells was increased by 30.8%, 22%, and 41.1%, respectively. After treatment with puromycin, the single cell viability of Bax^-/-, Bax-bak^-/-, and LDHa-Bax-bak^-/- cells was increased by 26.7%, 30.7%, and 38.8%, respectively. To further investigate the production performance of cells obtained after blocking apoptosis and lactic acid metabolism pathways, we induced transient expression of human tissue plasminogen activator (tPA) in these cells. The results showed that the secretion of tPA in Bax^-/-, Bax-Bak^-/-, and LDHa-Bax-Bak^-/- cells was 11.12%, 46.18%, and 63.13%, respectively, higher than that in wild-type CHO-K1 cells. The expression of intracellular tPA was increased by 35.65%, 130%, and 192.15%. In conclusion, blocking apoptosis and lactic acid metabolism pathways simultaneously can improve cell robustness and productivity, with the performance better than blocking the apoptosis pathway alone. The above results indicated that the constructed cell lines were expected to be the delivery carriers of protein drugs such as medicinal peptides, and better used for the treatment of diseases.
CHO Cells ; Cricetulus ; Animals ; Apoptosis/genetics* ; Lactic Acid/metabolism* ; Recombinant Proteins/biosynthesis* ; L-Lactate Dehydrogenase/genetics* ; bcl-2-Associated X Protein/genetics* ; bcl-2 Homologous Antagonist-Killer Protein/genetics* ; Cricetinae ; CRISPR-Cas Systems ; Staurosporine/pharmacology*

Objective To investigate the regulatory effect of poria cocos on gastrointestinal motility in mice. Methods A total of 130 Kunming mice were randomly divided into negative control group, low-dose and high-dose groups of raw poria cocos powder, low-dose and high-dose groups of cooked poria cocos powder, low-dose and high-dose groups of poria cocos surrogate culture powder, low-dose, medium-dose and high-dose groups of poria cocos water extract, and low-dose, medium-dose and high-dose groups of poria cocos alcohol extract, with 10 mice in each group. The animals were administered by gavage for 7 days, once a day. After the last administration, the intestinal propulsion function test and gastric solid emptying test were conducted to observe the regulating effect of poria cocos on gastrointestinal motility of mice. Results Compared with the negative control group, the small intestine propulsion rate in the low-dose group of poria cocos surrogate culture powder was significantly increased (P<0.01). Except the high-dose group of raw poria cocos powder, the other poria cocos groups had higher gastric residual rate (P<0.05). Conclusion Poria cocos does not promote intestinal propulsion of mice under normal physiological condition, but it can inhibit gastric empting and exert a moderating effect on gastrointestinal function in normal mice.

Objective:To extract rat brain microvascular endothelial cells(BMECs)using thin-layer cell culture method.Methods:The brain cortex of four-week-old SD rats was obtained,which was chopped,sieved,and digested with type II collagenase to obtain microvascular segments.The amount of culture medium was strictly controlled,and it was inoculated into culture flasks for primary culture.The target cells were morphologically observed by using an invert-ed phase contrast microscope,and the factor Ⅷ related antigen(FⅧRag)was identified by using immunocytochemi-cal staining.Results:The target cells cultured in the inverted phase contrast microscope showed typical endothelial-like monolayer cobble stone-like mosaic growth;FⅧRag positive cells accounted for more than 99％of the total cells.Conclusion:Thin-layer cell culture method can successfully isolate and cultivate high-purity rat BMECs.

Objective To evaluate the nutritional value of yeast protein, and to provide laboratory evidence for its application in the field of food and health food. Methods The amino acids content of yeast protein, soybean protein, whey protein and mixed protein was detected. Five nutritional evaluation methods, including amino acid score, ratio of total essential amino acid, ratio coefficient of amino acid, Pearson's correlation coefficient, and equally weighted gray correlation analysis were used to evaluate the protein quality. The protein efficacy test of the four proteins was performed in Sprague-Dawley rats to compare their nutritional value in vivo. Results The results showed that yeast protein was rich in essential amino acids, accounting for 47.58% of the total amino acids, and the ratio of essential amino acids to non-essential amino acids reached 0.91. Five nutritional evaluation methods all revealed that the yeast protein was an excellent protein in quality. The results of the protein efficacy test showed that yeast protein was able to promote the growth and development of the animals, and keep them in good nutritional condition, which was similar to whey protein, soy protein, and casein. The mixed protein had the highest corrected efficiency ratio among all the proteins tested in this study, possibly because the mixed ingredients made the amino acids complementary and thus had higher nutritional value. Conclusion Yeast protein can be used as a useful supplement to plant-derived and animal-derived protein products.

Objective Toinvestigatethevalueof"blendsign"onCTtopredictearlyhaematomaexpansioninacuteintracerebral haemorrhage(ICH).Methods SeventyGninepatientswithacuteICH whounderwentbaselineCTscanwithin6hourswereenrolled retrospectively.TheywerealsorecheckedwithCTscanin24hours.Allpatientsweredividedintoearlyhaematomaexpansiongroup and nonGhae m ato m a expansion group according to the change of hae m orrhage volu m e.M ultivariable L o g istic regression analysis w as usedtodetermineindependentriskfactorsofearlyhaematomaexpansion.Results Therewere28cases (35.4%)withhaematoma expansionin79patients."Blendsign"wasobservedin23patientsonbaselineCTscan,16of23 (69.6%)patientsappearedhaematoma expansion.Thesensitivity,specificity,positivepredictivevalue,negativepredictivevalueof"blendsign"forpredictingearlyhaematoma expansion w ere 57.1%,86.2%,69.6%,78.6%.M ultivariable L o g istic regression analysis sho w ed baseline hae m orrhage volu m e and"blendsign"wereindependentlyassociatedwithhaematomaexpansion.Conclusion "Blendsign"canbeusedtopredicthematoma expansioninacuteICH,whichishelpfultoidentifyhighriskpatientswithearlyhaematomaexpansiontomakethetreatmentmore promptlyandaccurately.

Objective To explore the effect of the standardized swallowing assessment (SSA) on extubation time for patients with nasal feeding. Methods One hundred patients with nasal feeding in our department from January 2013 to December 2014 were divided into the observation group and control group randomly with 50 in each group. The control group was treated with education on aspiratioin prevention and the right time for extubation was decided following doctor′s order. SSA was used in the observation group with weekly SSA and swallowing function, then the patients took pertinent functional exercises till a right time for extubation was decided. The two groups were compared in terms of incidences of re-intubation and aspiration pneumonia. Result The observation group had significantly lower incidences of re-intubation and aspiration pneumonia than the control one (P<0.01). Conclusions SSA is used to assess the swallowing function as well as the aspiration risks , which helps patients to take functional exercise and helps nurses to decide the right time for extubation. It can effectively lower the incidences of re-intubation and aspiration pneumonia and improve patients′prognosis.

AIM To explore the effect of long-term nebulized inhalation of nitroglycerin (Neb-NTG) on high pulmonary blood flow-induced pulmonary artery hypertension (PAH). METHODS 24 male Wistar rats were randomly divided into shunting group, Neb group and control group. An arteries-venous shunt was performed between the abdominal aorta and inferior vena cava shunting was produced in rats of Neb and shunt groups. Twelve weeks after the operation, nebulized nitroglycerin was inhaled by rats in the Neb group using pressurized air ejection mobilization, while nebulized saline was inhaled by rats of the other two groups. Ten minutes inhalation was given each day for each rat. After three weeks of inhalation, pulmonary artery systolic pressure (PASP) and mean pressure (PAMP) of each rat were evaluated by using a right cardiac catheterization procedure. Systemic arterial pressure (SAP) was reorded continuously by catheterization procedure. The ratio of right ventricular mass to body weight (RV/BW) and the ratio of right ventricular mass to left ventricular plus septal mass (RV/LV+S) were detected. Pulmonary vascular microstructure was measured. RESULTS PASP, RV/BW and RV/LV+S were significantly increased in shunt rats as compared with those of normal controls (P0.05). Muscularization of small pulmonary artery was obviously decreased in Neb-group compared with shunt group. CONCLUSION Long-term Neb-NTG ameliorates high pulmonary blood flow-induced PAH and pulmonary vascular remodeling.