Associations of genetic variations in pyroptosis related genes with acute adverse events in postoperative rectal cancer patients receiving concurrent chemoradiotherapy / 中华肿瘤杂志

Objective: This study aims to investigate the associations between genetic variations of pyroptosis pathway related key genes and adverse events (AEs) of postoperative chemoradiotherapy (CRT) in patients with rectal cancer. Methods: DNA was extracted from the peripheral blood which was collected from 347 patients before CRT. Sequenom MassARRAY was used to detect the genotypes of 43 haplotype-tagging single nucleotide polymorphisms (htSNPs) in eight pyroptosis genes, including absent in melanoma 2 (AIM2), caspase-1 (CASP1), caspase-4(CASP4), caspase-5 (CASP5), caspase-11 (CASP11), gasdermin D (GSDMD), gasdermin E (GSDME) and NLR family pyrin domain containing 3 (NLRP3). The associations between 43 htSNPs and AEs were evaluated by the odd ratios (ORs) and 95% confidence intervals (CIs) by unconditional logistic regression models, adjusted for sex, age, clinical stage, tumor grade, Karnofsky performance status (KPS), surgical procedure, and tumor location. Results: Among the 347 patients with rectal cancer underwent concurrent CRT with capecitabine after surgery, a total of 101(29.1%) occurred grade ≥ 2 leukopenia. rs11226565 (OR=0.41, 95% CI: 0.21-0.79, P=0.008), rs579408(OR=1.54, 95% CI: 1.03-2.29, P=0.034) and rs543923 (OR=0.63, 95% CI: 0.41-0.98, P=0.040) were significantly associated with the occurrence of grade ≥ 2 leukopenia. One hundred and fifty-six (45.0%) had grade ≥ 2 diarrhea, two SNPs were significantly associated with the occurrence of grade ≥ diarrhea, including CASP11 rs10880868 (OR=0.55, 95% CI: 0.33-0.91, P=0.020) and GSDME rs2954558 (OR=1.52, 95% CI: 1.01-2.31, P=0.050). In addition, sixty-six cases (19.0%) developed grade ≥2 dermatitis, three SNPs that significantly associated with the risk of grade ≥2 dermatitis included GSDME rs2237314 (OR=0.36, 95% CI: 0.16-0.83, P=0.017), GSDME rs12540919 (OR=0.52, 95% CI: 0.27-0.99, P=0.045) and NLRP3 rs3806268 (OR=1.51, 95% CI: 1.03-2.22, P=0.037). There was no significant difference in the association between other genetic variations and AEs of rectal cancer patients (all P>0.05). Surgical procedure and tumor location had great impacts on the occurrence of grade ≥2 diarrhea and dermatitis (all P<0.01). Conclusion: The genetic variants of CASP4, CASP11, GSDME and NLRP3 are associated with the occurrence of AEs in patients with rectal cancer who received postoperative CRT, suggesting they may be potential genetic markers in predicting the grade of AEs to achieve individualized treatment of rectal cancer.

Effects of neferine on invasion and migration of non-small cell lung cancer H1299 cells via inhibiting ROCK pathway / 中国药理学通报

Aim To observe the inhibitory effect of neferine(Nef)on the migration and invasion of non-small cell lung cancer(NSCLC)H1299 cells by blocking ROCK pathway.Methods H1299 cells were taken for in vitro culture, and treated with different concentrations of Nef.H1299 cell viability was measured by CCK-8 method to determine the dose of the experimental group.The migration and invasion abilities of H1299 cells were detected by cell scratch test and Transwell chamber test.The expression of matrix metalloproteinases MMP-2 and MMP-9 secreted from lung cancer cells was detected by enzyme linked immunosorbent assay(ELISA).The protein level of ROCK1 in H1299 cells was tested by real-time fluorescent quantitative PCR and Western blot; the binding mode and affinity between Nef and ROCK1 were stimulated by AutoDock semi flexible docking method.Results The doses of Nef in the experimental group were determined as 4, 6 and 10 μmol·L-1.These three concentrations of Nef could inhibit the migration and invasion of H1299 lung cancer cells to a certain degree in a dose-dependent manner.At the same time, Nef reduced the expression of MMP-2, MMP-9 and ROCK1 proteins related to the migration and invasion of the cancer cells.In addition, the affinity of Nef to ROCK1 was significantly higher than that of fasudil, an inhibitor of ROCK, and the binding force was stronger to A-chain of ROCK1.Conclusions As a potential natural anticancer compound, Nef can inhibit the migration and invasion of NSCLC by reducing the expression of MMP-2, MMP-9 and ROCK1 proteins related to the migration and invasion of the cancer cells.

Multimodal prehabilitation before major abdominal surgery: A retrospective study

INTRODUCTION@#Prehabilitation may benefit older patients undergoing major surgeries. Currently, its efficacy has not been conclusively proven. This is a retrospective review of a multimodal prehabilitation programme.@*METHODS@#Patients aged 65 years and above undergoing major abdominal surgery between May 2015 and December 2019 in the National University Hospital were included in our institutional programme that incorporated aspects of multimodal prehabilitation and Enhanced Recovery After Surgery concepts as 1 holistic perioperative pathway to deal with issues specific to older patients. Physical therapy, nutritional advice and psychosocial support were provided as part of prehabilitation.@*RESULTS@#There were 335 patients in the prehabilitation cohort and 256 patients whose records were reviewed as control. No difference in postoperative length of stay (@*CONCLUSION@#The current study found no differences in traditional surgical outcome measures with and without prehabilitation. An increase in patient mobility in the immediate postoperative period was noted with prehabilitation, as well as an association between prehabilitation and increased adherence to postoperative adjuvant therapy. Larger prospective studies will be needed to validate the findings of this retrospective review.

Influencing factors of serum calcium and the role of serum calcium on prognosis in sepsis patients / 中国医师进修杂志

Objective@#To retrospectively analyze the serum calcium level in sepsis patients, understand the influencing factors of abnormality calcium metabolism, and analyze the effect of serum calcium level on sepsis prognosis.@*Methods@#From January 1, 2017 to January 31, 2018, clinical data about sepsis patients admitted hospital were collected. The patients were divided into 2 groups according the levels of serum calcium measured in patients admitted in 24 h: normal serum calcium group (serum calcium 2.03-2.67 mmol/L), and low serum calcium group (serum calcium<2.03 mmol/L). And this study did not find hypercalcemia patients. The data about laboratory test index were compared between two groups.@*Results@#Fifty-two cases were included in this study, including 14 cases of normal serum calcium and 38 cases of hypocalcemia, and the incidence rate of hypocalcemia was 73.1%(38/52). The level of serum calcium in hypocalcemia group was (1.78 ± 0.17) mmol/L, and was (2.16 ± 0.14) mmol/L in normal serum calcium group. The main position of infections in patients with sepsis were digestive system, respiratory system and urinary system. Compared with that in the normal serum calcium group, the number of malnutrition patients in the hypocalcemia group was more. The total protein (TP), albumin (ALB), prealbumin (PAB) and total cholesterol (TCH) in the nutritional status of the patients were lower, the sequential organ failure assessment (SOFA) scores was higher, the prothrombin activity (PTA) was lower, the prothrombin time (PT) was prolonged, and international normalized ratio (INR) was increased. The differences were statistically significant (P<0.05 or <0.01). Pearson correlation test was used to screen the tests and scores related to the concentration of serum calcium. The results showed that the serum calcium concentration was negatively correlated with the level of SOFA scores, procalcitonin (PCT), PTA, thrombin time (PT), activated partial thromboplastin time (APTT) and INR (r=- 0.431, - 0.361, - 0.441, - 0.431, - 0.427, - 0.420, P<0.01 or<0.05), and the serum calcium concentration was positively correlated with TCH, TP, ALB and PAB (r=0.442, 0.475, 0.463, 0.419, P<0.01).@*Conclusions@#Hypocalcemia is related to the nutritional status of the body, and is related to coagulation function, serum protein level, PCT concentration and SOFA scores. The decrease of serum calcium indicates that multiple organ dysfunction syndrome may occur, and should be paid more attention to.

The effect of full nutritional management on nutrition status to on tumor patients with chemotherapy / 中国医师进修杂志

Objective To investigate the effect of full nutrition management on cancer patients with chemotherapy, and to support theory bases for the standardized clinical nutrition of cancer patients. Methods From January 2016 to December 2017, a total of 88 patients with the first chemotherapy were recruited and randomly divided into experimental group and control group, with 44 patients in each group. These patients all complied with the inclusion and exclusion criteria, and volunteered for the study. The two groups all received the same of anti-tumor treatment and the same conventional nutritional support (according to the results of nutrition screening and nutrition assessment, the nutritional support was formulated and patients′dietary guidance was given), while only patients of the experimental group were given full nutritional management, including regularly following up the patient′s condition changes, gastrointestinal reactions, and the feed. According to the five-step treatment model of malnutrition, nutritional support was improved in a timely manner, and appropriate nutritional support was provided. At the end of chemotherapy, the change of weight, albumin, prealbumin, hemoglobin was compared with that before treatment in the two groups. Results After chemotherapy, albumin, prealbumin, hemoglobin had no obvious drop compared with that before treatment in two groups (P>0.05). The patients of experimental group showed stable weight after chemotherapy: (53.39 ± 9.24) kg vs. (54.66 ± 9.41) kg, P>0.05. Weight loss in the control group after chemotherapy was statistically significant:(54.61 ± 10.76) kg vs. (56.52 ± 10.46) kg, P<0.05. Conclusions The full nutritional management can help maintain nutrition status during chemotherapy, especially can prevent the weight loss.

Effect of mesenchymal stem cell transplantation on immunological injury of the ovary in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of mesenchymal stem cell (MSC) transplantation in repairing ovarian injury in mice sensitized with porcine ovarian proteins.</p><p><b>METHODS</b>Wild-type female mice with ICR background (6-8 weeks old) were divided randomly into groups A, B and C (n=12). In groups B and C, the mice were treated with the total protein extract from porcine ovary to induce immunological injury of the ovary, while those in group A received no treatment. MSCs-derived from GFP transgenic mice were transplanted into the mice of group C, and equal volume of PBS was injected intraperitoneally in mice of the other two groups. PCR was used to detect GFP gene in the genomic DNA of the ovaries to assess MSCs homing in the ovary, and the reparative effect of MSCs on ovarian injury was evaluated using HE staining and TUNEL analysis.</p><p><b>RESULTS</b>After transplantation, the MSCs could reach the injured ovaries to promote the repair of the ovarian injury, resulting also in reduced apoptosis of the granulosa cells (GCs) in the injured ovaries.</p><p><b>CONCLUSION</b>MSCs transplantation can promote the recovery of the immunological injury of the ovary in mice, the mechanism of which may involve reduced apoptosis of the GCs.</p>

Effect of metformin on the formation of hepatic fibrosis in type 2 diabetic rats / 药学学报

The aim of this study is to investigate the effects of the metformin on the formation of hepatic fibrosis in type 2 diabetic rats and discuss its mechanism of liver-protecting activity. After SD rats were fed with high-fat and high-sucrose diet for four weeks, low-dose streptozotocin (STZ) was injected intraperitoneally to make the animal mode of type 2 diabetes. Then, all diabetic rats was fed with the high-fat diet and metformin (ig, 100 mg x kg(-1)) was given orally to metformin group for four months. After the last administration, fasting blood glucose was determined. The livers were removed to calculate the hepatic coefficient and to make HE and Picro acid-Sirius red staining, immunohistochemistry (alpha-SMA and TGFbeta1) and TUNEL staining in order to evaluate the effect of metformin on the hepatic fibrosis. The animal model of type 2 diabetes with hepatic fibrosis was successfully made. Metformin can significantly alleviate the lesions of hepatic steatosis and fibrosis, markedly reduce the expressions of alpha-SMA and TGFbeta1 in liver tissue of type 2 diabetic rats. However, TUNEL staining result suggested that metformin could not reduce apoptosis of hepatocytes. The results suggest that metformin can inhibit the formation of hepatic fibrosis in type 2 diabetes.

Effect of metformin on the formation of hepatic fibrosis in type 2 diabetic rats / 药学学报

The aim of this study is to investigate the effects of the metformin on the formation of hepatic fibrosis in type 2 diabetic rats and discuss its mechanism of liver-protecting activity. After SD rats were fed with high-fat and high-sucrose diet for four weeks, low-dose streptozotocin (STZ) was injected intraperitoneally to make the animal mode of type 2 diabetes. Then, all diabetic rats was fed with the high-fat diet and metformin (ig, 100 mg x kg(-1)) was given orally to metformin group for four months. After the last administration, fasting blood glucose was determined. The livers were removed to calculate the hepatic coefficient and to make HE and Picro acid-Sirius red staining, immunohistochemistry (alpha-SMA and TGFbeta1) and TUNEL staining in order to evaluate the effect of metformin on the hepatic fibrosis. The animal model of type 2 diabetes with hepatic fibrosis was successfully made. Metformin can significantly alleviate the lesions of hepatic steatosis and fibrosis, markedly reduce the expressions of alpha-SMA and TGFbeta1 in liver tissue of type 2 diabetic rats. However, TUNEL staining result suggested that metformin could not reduce apoptosis of hepatocytes. The results suggest that metformin can inhibit the formation of hepatic fibrosis in type 2 diabetes.

Secretory expression of recombinant porcine zona pellucida glycoprotein-3alpha (rpZP3alpha) in Pichia pastoris / 生物工程学报

To obtain the recombinant pZP3alpha protein for the study of the contraceptive vaccines, the DNA sequence (446-1423) encoding purified pZP3alpha was inserted into a vector--pPICZalphaA. The recombinant plasmid pPICZalphaA-pZP3alpha was linearized and then transformed into Pichia pastoris GS115 by electroporation. Engineering strains were attained by screening with zeocin and induced to produce rpZP3alpha in high-density fermentation. Then rpZP3alpha was purified by Cu2+ metal affinity column chromatography from the separated and concentrated fermentative supernatants. The purified rpZP3alpha was identified by SDS-PAGE and Western blot, and the quantity, purity and rate of recovery of the rpZP3alpha were analyzed by Quantity One software. One male rabbit was immunized with the Cu-NTA-purified rpZP3alpha. The antibody responses against rpZP3alpha and porcine ZP were detected by ELISA and the indirect immunofluorescence. Engineering strains expressing rpZP3alpha in secretion were constructed. A 46kD component named rpZP3alpha which can react with anti-pZP3 antibody was purified from fermentative supernatants of engineering strains and the average yield of purified rpZP3alpha obtained from fermentative supernatants was 8mg/L. The purity and the rate of recovery were up to 92% and 63% respectively. The anti-rpZP3alpha antiserum was prepared by immunization of a male rabbit with purified rpZP3alpha. This anti-rpZP3alpha antiserum could react with rpZP3alpha and purified pZP3 in ELISA and bind to porcine zona pellucida which produced bright green fluorescence in the indirect immunofluorescence. The rpZP3alpha (46kD) protein could be successfully expressed in the Pichia pastoris expression system. And this protein retained the immunogenic activity of natural pZP3.

Diagnosis and Treatment of Congenital Mesenteric Hiatual Hernia / 实用儿科临床杂志

Objective To explore the early diagnosis and treatment of congenital mesenteric hiatual hernia.Methods A retrospective study was carried out in 4 patients with congenital mesenteric hiatual hernia in Tongji hospital from Nov.2005 to Mar.2007,and combining lite-rature,the diagnosis and treatment of mesenteric hiatal hernia was summed up.Results Four patients were diagnosed in operation.One case was thought as adhesive intestinal obstruction before operation;two patients were on emergency operation and 2 patients were on time-elective operation;one patient preoperative CT scan may suggest mesenteric hiatal hernia;one case had partial resection of small intestinal,the others were replaced the intestine and fixed the defect.One patient occurred early septic shock;all of them had get well.Conclusions It′s hard to diagnose the congenital mesenteric hiatual hernia before operation.Abdomen CT examination and multislice spiral CT angiography (MSCTA) help to diagnose.Early diagnosis and timely operation are the therapeutic key of congenital mesenteric hiatual heria.For the patients with recurrent abdominal pain,who was not confirmed with a variety of inspection,laparoscopic exploration can provide diagnosis,and can take the initiative to control the development of disease.

Comparison of three methods for evaluating acrosome reaction in human spermatozoa / 中华男科学杂志

<p><b>OBJECTIVE</b>To find a convenient and exact method for evaluating acrosome reaction in human spermatozoa.</p><p><b>METHODS</b>The semen of the normal male was mixed and then divided into 6 groups. Coomassie brilliant blue (CBB) staining, chlortetracycline (CTC) fluorescence staining and acid phosphatase (ACP) detection were used for morphological observation and data analysis of the acrosome status of the human sperm treated with or without progesterone.</p><p><b>RESULTS</b>There were obvious morphological differences between the acrosome-reaction and acrosome-intact spermatozoa in CBB staining and CTC fluorescence staining, and significant differences were observed between the experimental and control spermatozoa by the three methods (P < 0.05).</p><p><b>CONCLUSION</b>All the three methods can be used to assess acrosome reaction in human spermatozoa, but Coomassie brilliant blue (CBB) staining is much more convenient and stable.</p>

Expression of recombinant human zona pellucida-3 protein (rhZP3) in Pichia pastoris / 生物工程学报

Human Zona Pellucida(ZP), which is a complex matrix surrounding oocytes,is comprised of three immunologically distinct glycoproteins(hZP1, hZP2 and hZP3). Because hZP3 possesses the sperm receptor activity and the acrosome-inducing activity, it has long been used as a candidate antigen to develop an immunocontraceptive vaccine. However, a large amount of native hZP3 protein is unavailable. It is an effective way to express hZP3 protein directly in vitro. Nevertheless, it had been reported that the rhZP3 protein produced in Pichia pastoris was not secreted but accumulated in the cells and could only be purified after being solubilized by strong denaturants. More unfortunately, after purification the final product required 6mol/L urea to maintain solubility. An improved project was advanced with the aim to express secreted and soluble rhZP3 protein in yeast. In this study, the fragment of hZP3 cDNA coding for aa 23 - 408, which the N-terminal leader was removed and most of the C-terminal transmembrane-like domain was reserved, was amplified by two PCR primers including EcoR I and Not I sites respectively and a His6 codon cassette was added to 5'-terminal. The hZP3 insert was incorporated into expression vector pPIC9K. The resulting recombinant yeast expression vector was designated pPIC9K-rhZP3. Linearized pPIC9K-rhZP3 was transformed into Pichia pastoris. After G418 selection, the recombinant Pichia pastoris strains were identified by PCR and the rhZP3 was expressed following the manufacturer' s protocol. Following induction with methanol, the rhZP3 protein was secreted and dissolved into the culture supernatant. SDS-PAGE and Western blot analyses showed that the apparent molecular weight of the expressed rhPZ3 proteins in yeast was smaller and a little size heterogeneity than native ones; after purified with Ni-chelating affinity chromatography, the final product's apparent molecular weight was about 32 - 34KD and their yield more than 20mg/L. We supposed that the C-terminal transmembrane-like domain be useful for secretion of rhZP3 into the culture supernatant and the expressed rhZP3 protein be incompletely digested by proteinases of Pichia into shorter fragments which all were glycosylated inhomogeneously. Fortunately, the fragments of rhZP3 protein can be recognized in Western blot by the polyclonal antibodies to porcine ZP3 which has showed a cross-reactivity with human ZP in vitro. It will be expected that the rhZP3 protein expressed in Pichia pastoris not only has immunogencity, say, it can rise antibodies in vivo to prevent spermatozoa-ovum binding, but also does not contain ovarian factors that might be the cause of undesired side effects, e.g. ovaritis and can be used as a safe immunogen in human antifertility vaccine research.