Screening and application of inducible promoters in Ruminiclostridium papyrosolvens.

Ruminiclostridium papyrosolvens is a promising candidate for producing renewable green chemicals from cellulose due to its cellulolytic and ethanologenic capabilities. It is of significance to screen effective, and convenient-to-use inducible promoters that can be used for regulating the gene expression in R. papyrosolvens. We characterized two endogenous inducible promoters and investigated another two exogenous ones on the adaptability in R. papyrosolvens. Both of the endogenous xylan-inducible promoter Pxyl and exogenous lactose-inducible promoter Plac are found of high specificity and stringency. Pxyl has a short time to be induced while Plac has a low concentration of inducer. With these findings, a mazF-based counter selectable system has been constructed for promoting the efficiency of mutant screening via plasmid curing. The inducible gene expression systems provided novel tools for enhancing the capability of genetic manipulation in engineering R. papyrosolvens. SIGNIFICANCE AND IMPACT OF THE STUDY: Four inducible promoters from Clostridia were characterized in R. papyrosolvens. Xylan-inducible promoter Pxyl was found of a short time while lactose-inducible promoter Plac needs a low concentration of inducer to induce. Employing them, we successfully construct a mazF-based counter selectable system, which would be used to increase the mutant screening efficiency via induction of plasmid curing. The inducible gene expression systems provided novel tools for enhancing the capability of genetic manipulation in engineering R. papyrosolvens.

Interleukin-armed chimeric antigen receptor-modified T cells for cancer immunotherapy.

Chimeric antigen receptor-modified T cells (CAR-T) are endowed with cytotoxic specificity to tumor cells. Although CAR-T-based cancer immunotherapy presents curable therapeutic potential for hematological malignancies, achieving substantial efficacy for solid tumors remain challenging. Researchers have exploited many strategies to enhance the anti-tumor efficacy of CAR-T cells for solid tumors, among which cytokine-armed CAR-T cells improve the proliferation, survival, homing and other properties of CAR-T cells. Interleukins (ILs), pivotal cytokines that affect the function of immune cells, were co-expressed in CAR-T cells or combinatorially administered to enhance the therapeutic potential in clinical trials. In this review, we summarize the strategies exploited by ILs to improve the anti-cancer ability of CAR-T cells and the different impacts of different ILs on CAR-T cells.

Effects of Pax3 and Pax7 expression on muscle mass in the Pekin duck (Anas platyrhynchos domestica).

This study aimed to investigate whether the differential expression of muscle development-related genes is one of the reasons why muscle development differs between Pekin, Jianchang, and Heiwu ducks, which are all domesticated duck breeds (Anas platyrhynchos domestica) breeds. At 2 weeks of age, the RNA expression of paired box 7 (Pax7), paired box 3 (Pax3), myogenic differentiation antigen (MYOD), and myogenin (MYOG) genes were measured by quantitative polymerase chain reaction, and Pax3 and Pax7 protein levels were detected by western blot assay. Myofiber morphology was investigated using paraffin-embedded muscle sections. At 8 weeks of age, 30 ducks of each breed were slaughtered for meat quality determination. The results revealed that Pax3 and Pax7 expression levels at both the RNA and protein levels were high in the Pekin duck. In addition, MYOG expression levels in the Jianchang duck were significantly higher than in the other two duck breeds (P < 0.05). There were no significant differences in MYOD expression levels between the breeds (P > 0.05). Myofiber diameter and cross-sectional area were the largest in the Pekin duck and the smallest in the Heiwu duck. There were significant differences in slaughter data between these breeds, and muscle content was greatest in the Pekin duck. The results indicate that the muscle content of three different duck breeds is associated with the expression of satellite-cell marker genes.

Characterization and molecular mapping of stripe rust resistance gene Yr61 in winter wheat cultivar Pindong 34.

KEY MESSAGE: We report a new stripe rust resistance gene on chromosome 7AS in wheat and molecular markers useful for transferring it to other wheat genotypes. Several new races of the stripe rust pathogen have established throughout the wheat growing regions of China in recent years. These new races are virulent to most of the designated seedling resistance genes limiting the resistance sources. It is necessary to identify new genes for diversification and for pyramiding different resistance genes in order to achieve more durable resistance. We report here the identification of a new resistance gene, designated as Yr61, in Chinese wheat cultivar Pindong 34. A mapping population of 208 F2 plants and 128 derived F2:3 lines in a cross between Mingxian 169 and Pindong 34 was evaluated for seedling stripe rust response. A genetic map consisting of eight resistance gene analog polymorphism (RGAP), two sequence-tagged site (STS) and four simple sequence repeat (SSR) markers was constructed. Yr61 was located on the short arm of chromosome 7A and flanked by RGAP markers Xwgp5467 and Xwgp5765 about 1.9 and 3.9 cM in distance, which were successfully converted into STS markers STS5467 and STS5765b, respectively. The flanking STS markers could be used for marker-assisted selection of Yr61 in breeding programs.

[Retrospective analysis on 77 cases of T4b hypopharyngeal carcinoma treated by non-surgical treatments].

Objective: To analyze the effectiveness, safety and factors influencing the clinical prognosis of patients with hypopharyngeal carcinoma in T4b by nonsurgical treatments. Methods: The clinical data of 77 patients with T4b hypopharyngeal cancer treated in the College of Otolaryngology Head and Neck Surgery of the Chinese People's Liberation Army General Hospital from January 2010 to June 2021 were analyzed retrospectively. All were males, aged(57.0±8.0)years old. Patients were treated with induction chemotherapy plus concurrent chemoradiotherapy. Kaplan Meier survival analysis was used to compare the effects of different factors on prognosis. Adverse reactions during treatments and the causes of death were analyzed. Results: 98.7% of 77 patients with T4b hypopharyngeal cancer completed the chemotherapy plan and 94.8% completed the radiotherapy plan. The most common adverse reactions were grade 2 radiation oral mucositis (50/77, 64.9%) and grade 2 leukopenia (50/77, 64.9%). The incidence of grade 3 severe hoarseness was 7.8% (6/77), one patient (1.3%) underwent gastrostomy due to dysphagia, and pronunciation and swallowing function were effectively preserved in other patients. The overall survival rate was 71.9% at 1 year, 45.6% at 3 years and 29.7% at 5 years. The location of tumor, the presence of liquefaction necrosis in tumor, the use of molecular targeted drugs and the approach of radiotherapy were independent factors,each of which that affected the prognosis of T4b patients with advanced hypopharyngeal cancer [HR (95%CI) were 1.867(1.085-3.213), 3.018 (1.437-6.335), 0.372 (0.181-0.764) and 2.158 (1.015-4.588), respectively, P<0.05]. The two leading causes of death with high incidence were disease recurrence (12/32, 37.5%) and cervical large vessel rupture and hemorrhage (11/32, 34.4%). Conclusions: Non-surgical comprehensive treatment offers a high laryngeal preservation rate in patients with T4b hypopharyngeal cancer. The location of tumor, the liquefaction necrosis within tumor, the use of molecular targeted drugs, and the approach of radiotherapy are independent prognostic factors.

Protective effect of Bu-Zhong-Yi-Qi decoction, the water extract of Chinese traditional herbal medicine, on 5-fluorouracil-induced intestinal mucositis in mice.

Intestinal mucositis is a serious toxic side effect of 5-fluorouracil (5-FU) treatment. Bu-Zhong-Yi-Qi decoction (BZYQD), a water extract of Chinese traditional herbal medicine, is widely used in chemotherapy in Asia as an alternative treatment to reduce the side effects of chemotherapy. However, the mechanism is unknown. To evaluate its mechanism, we investigated the effect of BZYQD on 5-FU-induced intestinal mucositis in mice, especially with regard to apoptosis in the intestinal mucosal epithelia. In the present study, mice were divided into three groups: control, 5-FU, and 5-FU + BZYQD. Mice in the 5-FU and 5-FU + BZYQD groups were administered 5-FU (100 mg/kg/day, intraperitoneally) for 6 days, and the mice in the latter group were given BZYQD (8 g/kg/day, intragastrically) beginning 4 days before 5-FU and continuing until the termination of the experiment. Loss in body weight and diarrhea during the 5-FU treatment were significantly attenuated by administration of BZYQD. The morphological signs of intestinal damage, including shortened villi height, crypt destruction, apoptosis, and necrosis, in intestinal mucosal epithelia were also reversed, accompanied by reduced neutrophil infiltration, nitrite levels, and inflammatory factors (tumor necrosis factor α and interleukin 1ß) and increased levels of reduced glutathione. These results suggest that BZYQD inhibits 5-FU-induced intestinal mucositis, and this effect may be due to the reduction in apoptosis and necrosis in intestinal mucosal epithelia via the suppression of inflammatory cytokine upregulation. In conclusion, inhibiting cytokine-mediated apoptosis or necrosis can be the molecular mechanism by which BZYQD reduces the gastrointestinal side effects of cancer chemotherapy.

An in vivo and in vitro study: High-dosage Danshen injection induces peripheral vascular endothelial cells injury.

Danshen injection, a pharmaceutical dosage form of Danshen, has been widely used in the treatment of coronary heart diseases, myocardial infarction, and hypertension. With more and more adverse drug reactions linked with Danshen injection, its safety comes under suspicion. To evaluate its safety, mice were divided into four groups: vehicle, low-, middle-, and high-Danshen group, and each group was intravenously administered with Danshen injection at a dose of 0, 0.64, 1.55, and 5.76 g/kg/day for 5 days, respectively (the low dosage was the recommended clinical dosage, the middle dosage was the most commonly used higher dosage, and the high dosage was the highest dosage used in clinic). Peripheral vascular toxicity wasn't observed in the low-dosage group, elevated serum endothelin-1 (ET-1) was observed in the middle-dosage group; and more peripheral vascular toxicities like increased vascular leakage, elevated serum nitrate and ET-1, and vascular endothelial cells apoptosis were detected in the high-dosage group. In vitro study, low-concentration Danshen injection showed protective effect to human umbilical vein endothelial cells (HUVECs), while high concentration displayed strong cytotoxic effects, including increase in nitric oxide and ET-1 production, inhibition of cell viability, and apoptosis induction. Further, the HUVECs' apoptosis induced by high-concentration Danshen injection was found along with the induction of reactive oxygen species. In conclusion, these results suggest that Danshen injection is nontoxic in its recommended clinical dosage, and the 2.4-fold as the recommended clinical dosage might be the highest safety dosage in clinic treatment. In addition, Danshen injection is a potential vascular toxic drug in its high dosage and shouldn't be used far beyond its recommended dosage in clinic treatment.

Adjuvant intensity-modulated radiotherapy (IMRT) with concurrent paclitaxel and cisplatin in cervical cancer patients with high risk factors: A phase II trial.

OBJECTIVE: To evaluate the safety and efficacy of adjuvant intensity-modulated radiotherapy (IMRT) with concurrent paclitaxel and cisplatin (TP) in early stage cervical cancer patients with high risk factors after radical hysterectomy. METHODS: Patients who underwent radical hysterectomy for FIGO stage IB-IIA cervical cancer and had high risk factors for recurrence were recruited. One cycle of TP was delivered before and after concurrent chemoradiotherapy, respectively. Concurrent chemoradiotherapy began 21 days after the start of the initial cycle of the chemotherapy with two cycles of TP delivered on day 1 and day 29 of radiotherapy. Primary endpoints were overall survival (OS) and relapse-free survival (RFS), with toxicities, local-regional control (LC) and distant failure (DF) rate as secondary endpoints. RESULTS: Between 2008 and 2012, 67 patients were evaluable. The 2 and 4-year RFS rates were 98.2% and 92.9%. Corresponding OS rates were 100%, and 98.0%, respectively. The 4-year LC and DF rates were 98.0% and 5.2%, respectively. Grade 3-4 acute leucopenia, neutropenia and thrombocytopenia occurred in 25.4%, 11.9% and 1.5% of patients, respectively. There were 89.6% and 59.7% patients experienced acute vomiting and diarrhea, but only 6.0% and 6.0% patients were grade 3, respectively. No case of chronic toxicity exceeded grade 2. CONCLUSION: Adjuvant concurrent IMRT with paclitaxel plus cisplatin are safe and effective in early stage cervical cancer patients with high risk factors for recurrence following radical hysterectomy.

Role of urbanization and air pollution in adolescent asthma: a mass screening in Taiwan.

BACKGROUND AND PURPOSE: The prevalence of asthma in school children in Taiwan is increasing. This study used mass screening among middle school children in Taiwan to determine the prevalence of asthma and related factors. METHODS: Data were collected from parents using a self-reported questionnaire and from children using the International Study of Asthma and Allergies in Childhood (ISAAC) video questionnaire. Six study teams conducted the survey nationwide in 1995-1996, with the assistance of middle school nurses and teachers. RESULTS: Among the 1,018,031 students at 795 middle schools who returned questionnaires, 8.5% had a history of asthma (ranging in prevalence from 4.2% to 13% in 25 areas). The prevalence of asthma was higher in boys than in girls (10.0% vs 7%) and was highest in more urbanized areas (11.2%), followed by moderately urbanized areas (7.4%) and less urbanized and rural areas (6.5%). Controlling for age, family smoking, family incense burning, and parental education level, multivariate logistic regression models indicated that children living in an area with heavy air pollution were more likely to have asthma than those in an area with no or light pollution (odds ratio, OR = 2.01 and 95% confidence interval, CI = 1.94-2.09 based on parental ranking of pollution level, or OR = 1.30 and 95% CI = 1.18-1.42 based on pollution level reported by the Environmental Protection Administration). CONCLUSIONS: Adolescent asthma in Taiwan is most prevalent in the most urbanized areas and decreases in prevalence in less urbanized areas. This study also found that higher parental education level and higher area air pollution were associated with higher adolescent asthma prevalence.

[Successful local adoptive immunotherapy for pleuritis carcinomatosa due to renal cell cancer. A case report].

A 54-year-old woman who had undergone radical nephrectomy for renal cell cancer nine months before was admitted to our hospital because of difficulty in breathing. X-ray films of the chest showed massive pleural effusion on the left side and cytological examination of the effusion revealed malignant cells which may have originated from renal cell cancer. Intrapleural instillations of interleukin-2 and of tumor infiltrating lymphocytes isolated from the pleural effusion were performed. After the initiation of the treatment, the pleural effusion decreased and malignant cells disappeared from the pleural fluid. Partial response defined by the criteria provided by the Japan Lung Cancer Society was achieved. No serious side effects were observed. This would be a useful treatment for pleuritis carcinomatosa by renal cell cancer.

[Determination of organochlorine pesticide multi-residues in Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao, Panax notoginseng (Burk.) F. H. Chen and P. quin quefolium L].

OBJECTIVE: To determine 15 organochlorine pesticide residues in Astragalus membranaceus, Panax notoginseng and P. quinquefolium. METHODS: The 15 organochlorine pesticides were extracted from samples with mixed solvents by ultrasonication, cleaned up by a column packed with Florisil, and then separated by capillary gas chromatography with electron capture detector(CGC-ECD). The capillary column was DB-5(15 m x 0.32 mm x 0.25 microns); injector temperature 240 degrees C; detector temperature 320 degrees C; the column temperature program initial 100 degrees C (hold 0.5 min), rate of 20 degrees C min-1 to 210 degrees C (hold 0.5 min), rate of 8 degrees C min-1 to 240 degrees C (hold 10 min). The column head-pressure was 58.84 kPa. The injection was splitless and the injection volume was 1 microliter. RESULT AND CONCLUSION: The average recoveries and RSD are in the range of 78.30%-130.1% and 0.75%-31.9% at two spiked mixed organochlorine pesticide levels, respectively. All the samples have organochlorine pesticide residues.

Circadian rhythm of serum and lymph apolipoprotein AIV in ad libitum-fed and fasted rats.

The aim of the present study was to determine if there is a circadian rhythm in serum and lymph apolipoprotein (apo) AIV and what factors determine this rhythm. Male Sprague-Dawley rats with chronic right atrial catheter were housed in a room illuminated from 0600 to 1800. With ad libitum feeding, serum apo AIV concentration showed a circadian rhythm concomitant with the feeding pattern. In 24-h fasted rats, the serum apo AIV concentration maintained a circadian rhythm and was high during the dark. With mesenteric lymph diversion, serum apo AIV concentration diminished and the circadian rhythm was abolished. The lymph flow, lymph apo AIV, cholesterol, triacylglycerol, and phospholipid contents all exhibited the same circadian rhythm in fasting, with the levels higher in the dark. These circadian rhythms were abolished after bile diversion. In conclusion, serum apo AIV in ad libitum-fed and fasted rats exhibits a circadian rhythm governed by lymph apo AIV output. Furthermore, bile was an important determinant of the circadian rhythm of lymph flow, lymph apo AIV, triacylglycerol, cholesterol, and phospholipid output.