[Effects of Microplastic High-density Polyethylene on Cotton Growth, Occurrence of Fusarium wilt, and Rhizosphere Soil Bacterial Community].

Plastic mulch, especially polyethylene mulch, is widely used in agricultural production in China, but the microplastics formed by its degradation gradually have accumulated in soil, causing a series of environmental problems. At present, there have been many reports on the environmental biological effects of microplastics in farmland soil, but studies on the effects of microplastics on crop growth, disease occurrence, and rhizosphere soil bacterial communities are still lacking. In the previous study, it was found that 1% high-density polyethylene (HDPE, 500 mesh) could increase the incidence rate of cotton Fusarium wilt (33.3%) and inhibit growth, but this phenomenon was not found after soil sterilization. It was speculated that HDPE could affect the growth and occurrence of Fusarium wilt by regulating the soil microbial community. Therefore, high-throughput sequencing technology, combined with network and FAPROTAX function analysis, were used to investigate the effects of HDPE on the bacterial community structure, interaction network, and soil function in cotton rhizosphere in order to analyze the mechanism of HDPE. NovaSeq sequencing showed that the bacterial community of HDPE-treated cotton rhizosphere soil was composed of 54 phyla and 472 genera; the number of phyla and genera was higher than that in untreated soil. The α and ß diversity and ANOSIM/Adonis analyses showed that HDPE significantly reduced the richness of the bacterial community and changed the composition of the community structure. Based on a T-test species difference analysis, HDPE significantly reduced the relative abundance of bacteria with biological control, pollutant degradation, and antifungal drug synthesis (such as Kribbella, Massiliam, Hailiangium, and Ramlibacter).The change in the bacterial community will lead to the change in soil bacterial function. Further analysis of FAPROTAX function revealed that HDPE weakened some biochemical functions of bacteria in the cotton rhizosphere soil, such as aerobic chemoheterotrophy, fermentation, and nitrate reduction. The correlation network at the genus level showed that HDPE treatment weakened the interaction between rhizosphere bacteria, reduced the number of positive correlation connections, increased the number of negative correlation connections, simplified network structure, and changed the key flora. The above results showed that HDPE could reduce the cotton growth and the occurrence of Fusarium wilt by changing the bacterial community, interaction, and functional metabolism in rhizosphere soil, which can provide guidance for evaluating the ecological risk of polyethylene microplastics and the remediation of contaminated soil.

Discovery of Novel 3,11-Bispeptide Ester Arenobufagin Derivatives with Potential in Vivo Antitumor Activity and Reduced Cardiotoxicity.

Arenobufagin, one of the bufadienolides isolated from traditional Chinese medicine Chan'su, exhibits potent antitumor activity. However, serious toxicity and small therapeutic window limits its drug development. In the present study, to our knowledge, novel 3,11-bispeptide ester arenobufagin derivatives have been firstly designed and synthesized on the base of our previous discovery of active 3-monopeptide ester derivative. The in vitro antiproliferative activity evaluation revealed that the moiety at C3 and C11 hydroxy had an important influence on cytotoxic activity and selectivity. Compound ZM350 notably inhibited tumor growth by 58.8 % at a dose 10 mg/kg in an A549 nude mice xenograft model. Therefore, compound ZM350 also presented a concentration-dependent apoptosis induction and low inhibitory effect against both hERG potassium channel and Cav1.2 calcium channel. Our study suggests that novel 3,11-bispeptide ester derivatives will be a potential benefit to further antitumor agent development of arenobufagin.

Identification of genetic variants of the IL-22 gene in association with an altered risk of COPD susceptibility.

The incidence of chronic obstructive pulmonary disease (COPD) is related to the interaction between environmental exposure and genetic factors. Far more than 15% of smokers eventually develop COPD. In addition to smoking, genetic susceptibility may be another factor in the development of COPD. IL-22 and its receptors are increased in human and experimental COPD and contribute to pathogenesis. Here, we conducted a case-control study to evaluate the association between IL-22 tag-single nucleotide polymorphisms (SNPs) and COPD risk. Four tag-SNPs (rs2227478, rs2227481, rs2227484 and rs2227485) were identified according to linkage disequilibrium (LD) analysis in 30 healthy controls. A total of 513 COPD cases and 504 controls were recruited to perform an association study between these four tag-SNPs and COPD risk. We found that the "C" allele of rs2227478T>C and the "T" allele of rs2227481C>T were obviously related to decreased COPD susceptibility. Genetic model analysis showed that rs2227478T>C and rs2227481C>T were significantly associated with a decreased risk of COPD under dominant models after adjusting for the above factors. In the recessive model, rs2227485T>C was obviously associated with decreased COPD risk. Our data showed that only rs2227485T>C was associated with a decreased COPD risk after Bonferroni correction. The eQTL analysis showed that rs2227485T>C was significantly associated with IL-22 expression. The pGL4-rs2227485-C gene reporter had a higher promoter activity than pGL4-rs2227485-T. In our study, rs2227485T>C, located in the promoter region of IL-22, was associated with a decreased risk of COPD and increased IL-22 promoter activity, suggesting that this variant might modulate COPD susceptibility.

[The Role of Platelet Exosomes in Atherogenic Thrombosis --Review].

Exosomes are subtypes of extracellur vesicles containing a variety of cell-specific proteins, lipids and nucleic acids released during cell activation or apoptosis, and play the role of intercellur communication mediators in different physiological and pathological processes. With the development of research in recent years, the role of platelet-derived exosomes in cardiovascular diseases has attracted extensive attention. This paper reviews the role of platelet-derived exosomes in atherosclerotic thrombosis and the potential role of platelet-derived exosomes as biomarkers for the diagnosis and treatment of atherosclerotic thrombotic disease and the problems to be solved.

Gut Microbial Characteristics of Adult Patients With Epilepsy.

OBJECTIVE: To characterize the intestinal flora of patients with epilepsy and its correlation with epilepsy. METHODS: Patients with ages > 18 years were consecutively enrolled from the outpatient department, Affiliated Hospital of Guizhou Medical University from January 2018 to December 2019. A total of 71 subjects were recruited, including epilepsy patients (n = 41) as an observation group and patient family members (n = 30) as a control group. Fresh stool specimens of all the subjects were collected. The 16S ribosomal RNA sequencing was analyzed to determine changes in intestinal flora composition and its correlation with epilepsy. Subgroup analysis was then conducted. All patients with epilepsy were divided into an urban group (n = 21) and a rural group (n = 20) according to the region, and bioinformatics analyses were repeated between subgroups. RESULTS: LEfSe analysis showed that Fusobacterium, Megasphaera, Alloprevotella, and Sutterella had relatively increased abundance in the epilepsy group at the genus level. Correlation analysis suggested that Fusobacterium sp. (r = 0.584, P < 0.01), Fusobacterium mortiferum (r = 0.560, P < 0.01), Ruminococcus gnavus (r = 0.541, P < 0.01), and Bacteroides fragilis (r = 0.506, P < 0.01) were significantly positively correlated with the occurrence of epilepsy (r ≥ 0.5, P < 0.05). PICRUSt function prediction analysis showed that there were significant differences in 16 pathways between the groups at level 3. Comparing the rural group with the urban group, Proteobacteria increased at the phylum level and Escherichia coli, Fusobacterium varium, Prevotella stercorea, and Prevotellaceae bacterium DJF VR15 increased at the species level in the rural group. CONCLUSION: There were significant differences in the composition and functional pathways of gut flora between epilepsy patients and patient family members. The Fusobacterium may become a potential biomarker for the diagnosis of epilepsy.

The Role of Platelet Exosomes in Atherogenic Thrombosis --Review / 中国实验血液学杂志

Exosomes are subtypes of extracellur vesicles containing a variety of cell-specific proteins, lipids and nucleic acids released during cell activation or apoptosis, and play the role of intercellur communication mediators in different physiological and pathological processes. With the development of research in recent years, the role of platelet-derived exosomes in cardiovascular diseases has attracted extensive attention. This paper reviews the role of platelet-derived exosomes in atherosclerotic thrombosis and the potential role of platelet-derived exosomes as biomarkers for the diagnosis and treatment of atherosclerotic thrombotic disease and the problems to be solved.

A case report of brain abscess caused by Nocardia farcinica.

BACKGROUND: Brain abscess due to the Nocardia genus is rarely reported and it is usually found in immunocompromised patients. Treatment of Nocardia brain abscess is troublesome and requires consideration of the severity of the underlying systemic disease. The difficulties in identifying the bacterium and the frequent delay in initiating adequate therapy often influence the prognosis of patients. CASE PRESENTATION: Here, we report a rare case of brain abscess caused by Nocardia farcinica. The patient's medical history was complicated: he was hospitalized several times, but no pathogens were found. At last, bacteria were found in the culture of brain abscess puncture fluid; the colony was identified as Nocardia farcinica by mass spectrometry. Targeted antibiotic treatment was implemented, brain abscess tended to alleviate, but the patient eventually developed fungal pneumonia and died of acute respiratory distress syndrome (ARDS). CONCLUSION: Brain abscess caused by Nocardia farcinica can appear in non-immunocompromised individuals. Early diagnosis, reasonable surgical intervention, and targeted antibiotic treatment are critical for Nocardia brain abscess treatment. In the treatment of Nocardia brain abscess, attention should paid be to the changes in patients' immunity and infection with other pathogens, especially fungi, avoided.

Luteolin inhibits the proliferation, adhesion, migration and invasion of choroidal melanoma cells in vitro.

Choroidal melanoma is a devastating disease that causes visual loss and a high mortality rate due to metastasis. Luteolin, a potential anticancer compound, is widely found in natural plants. The aim of this study was to evaluate the antiproliferative, antiadhesive, antimigratory and anti-invasive effects of luteolin on choroidal melanoma cells in vitro and to explore its potential mechanism. Cell counting kit-8 (CCK-8) assays, 5-ethynyl-2'-deoxyuridine (EdU) assays, Cell adhesion, migration, and invasion assays were performed to examine the inhibitory effects of luteolin on cell cell viability, proliferation, adhesion, migration and invasion capacities, respectively. Considering the correlation between Matrix metalloenzymes and tumor metastasis, Enzyme-linked immunosorbent assays (ELISAs) were used to assess matrix metalloproteases MMP-2 and MMP-9 secretion. Western blotting was performed to detect p-PI3K P85, Akt, and p-Akt protein expression. The cytoskeletal proteins vimentin were observed with cell immunofluorescence staining. Luteolin can inhibit OCM-1 cell proliferation, migration, invasion and adhesion and C918 cell proliferation, migration, and invasion through the PI3K/Akt signaling pathway. Therefore, Luteolin may have potential as a therapeutic medication for Choroidal melanoma.

LncRNA MIAT Promotes Inflammation and Oxidative Stress in Sepsis-Induced Cardiac Injury by Targeting miR-330-5p/TRAF6/NF-κB Axis.

Sepsis is a whole-body inflammation and main cause of death in intensive care units worldwide. We aimed to investigate the roles of lncRNA MIAT and miR-330-5p in modulating inflammatory responses and oxidative stress in lipopolysachariden (LPS)-induced septic cardiomyopathy. Serum and heart tissue were collected from in vivo septic mice model, ELISA and qRT-PCR were used to measure the expression of pro-inflammation cytokines, MIAT and miR-330-5p, respectively. The knockdown of MIAT and overexpression of miR-330-5p were conducted to assess their effects on regulating inflammation response and intracellular oxidative stress in LPS-stimulated HL-1 cells. The reactive oxygen (ROS) level, mitochondrial membrane potential (MMP), GSH/GSSH ratio, and lipid peroxidation assessment (MDA) were used to evaluate the intracellular oxidative stress. Dual-luciferase reporter assay was performed to identify the association between MIAT and miR-330-5p, TRAF6 and miR-330-5p, respectively. In septic mice, the expression of MIAT and pro-inflammation cytokines was elevated while the expression of miR-330-5p decreased. Knockdown of MIAT or overexpression of miR-330-5p restrained inflammation and oxidative stress induced by LPS in vitro; MIAT directly targeted miR-330-5p to regulate NF-κB signaling, and miR-330-5p targeted against TRAF6 to suppress the activation of NF-κB signaling. We determined that lncRNA MIAT directly binds to miR-330-5p to activate TRAF6/NF-κB signaling axis and further promotes inflammation response as well as oxidative stress in LPS-induced septic cardiomyopathy. This finding suggests the potential therapeutic role of lncRNA MIAT and miR-330-5p in LPS-induced myocardial injury.

Spinal Clear Cell Meningiomas: Clinical Features and Factors Predicting Recurrence.

OBJECTIVE: As a World Health Organization grade II tumor of the nervous system, clear cell meningioma (CCM) is an uncommon histologic variant of meningioma. Spinal CCMs are even rarer, with <100 spinal CCMs reported in the English literature. We present this study to characterize clinical manifestations of spinal CCMs and determine the factors predicting recurrence. METHODS: A literature search was performed for relevant case reports and series in PubMed and Embase until September 1, 2019. These articles were reviewed to identify clinical features, treatment modalities, and prognosis of patients with spinal CCMs. RESULTS: Eighty-four spinal CCMs were analyzed. Of these patients, 36 (42.9%) were young (age ≤18 years), and the mean age at resection was 24 years. Fifty-three patients (63.1%) were female and 31 (36.9%) were male. Most of the tumors (56/84, 66.7%) were located in the lumbar region. In 31 patients (36.9%) >2 segments in the craniocaudal direction were involved (number of involved segments ≥3 levels). Gross tumor resection was performed in 77 patients (91.7%). Twenty patients (23.8%) showed radiographic evidence of recurrence during follow-up. Recurrence-free survival at 1, 5, and 10 years after resection of spinal CCM was 87%, 71%, and 47%, respectively. Multivariate analysis showed that age ≤18 years (hazard ratio [HR], 3.64; P = 0.024), subtotal resection (HR, 3.43; P = 0.031), and segments involving ≥3 levels (HR, 5.66; P = 0.002) were associated with increased recurrence. CONCLUSIONS: Spinal CCMs have their own unique clinical features compared with conventional spinal meningiomas and intracranial CCMs. Spinal CCMs have a predilection to affect younger patients, are prone to appear in the lumbar region, and have a high recurrence rate. Age ≤18 years, subtotal resection, and involvement of long segments (≥3 levels) are positive predictors of recurrence.

Factors Affecting the Prognosis of Women with Liver Dysfunction in the Third Trimester of Pregnancy / 中山大学学报(医学科学版)

@#【Objective】To explore the effects of liver dysfunction in the third trimester of pregnancy on maternal outcomes and identify the factors affecting the maternal prognosis.【Methods】We collected the clinical data of 1 113 women with liver dysfunction in the third trimester of pregnancy （case group） and 1 113 normal pregnancies （control group） from the Third Affiliated Hospital of Sun Yat-sen University between January 2015 and December 2018. We compared the rates of cesarean section，premature delivery，placental abruption，postpartum hemorrhage and maternal mortality in the two groups，conducted the univariate and multivariate analysis for the case group and determined the prognostic risk factors by using Logistic regression. Receiver operating characteristic（ROC）curve analysis was applied to estimate the value of each independent risk factor for predicting liver dysfunction-related maternal mortality. 【Results】The rates of cesarean section，premature delivery，placental abruption，postpartum hemorrhage in the case group were higher than those in the control group（P < 0.05），and the odds ratios（ORs）were 3.59 ，7.81 ，10.68 and 2.93 ，respectively. The maternal mortality in the case group（1.2%）was higher than that in the control group（0.0%）（P < 0.05）. Logistic analysis revealed that high total bilirubin（TBIL），low prothrombin activity（PTA）and low fasting plasma glucose（FPG）were independent risk factors for liver dysfunction- related maternal mortality. The ROC curve analysis indicated that when TBIL was 235.4 μmol/L，the Youden′ s index in maximum was 0.331 with sensitivity of 0.818 and specificity of 0.513. When PTA was 20.5% ，the Youden′ s index in maximum was 0.366 with sensitivity of 0.821 and specificity of 0.545. When FPG was 3.11 mmol/L，the Youden′s index in maximum was 0.405 with sensitivity of 0.769 and specificity of 0.636.【Conclusion】Liver dysfunction in the third trimester of pregnancy has adverse effects on maternal outcomes. TBIL ，PTA and FPG are the factors affecting the maternal prognosis and may have certain predictive value for maternal death.

Effect of Jiajian Fuyuan Huoxuetang Combined with Electroacupuncture on Neurological Rehabilitation After Spinal Cord Injury / 中国实验方剂学杂志

Objective:To observe the effect of Jiajian Fuyuan Huoxuetang combined with electroacupuncture on neurological rehabilitation of patients with spinal cord injury, and to investigate its effect on neurotrophic and inflammatory factors. Method:One hundred and twenty patients were randomly divided into control group (59 cases) and observation group (61 cases) by random number table. Both groups’ patients got monosialotetrahexosylganglioside by intravenous drip for 6 weeks, 200-400 mg/time, 1 time/day.Rat nerve growth factor for injection by intramuscular injection for 4 weeks, 20 μg/time, 1 time/day. Patients in control group additionally got electroacupuncture treatment,and tongluo huatan capsule,3 caps/time,3 time/days. The patients in observation group additionally got Jiajian Fuyuan Huoxuetang combined with electroacupuncture, 1 dose/day. The courses of treatment were 12 weeks in two groups. Scores of degree of spinal cord neurological impairment were graded by using impairment score of American Spinal Injury Association (ASIA). Lower extremity motor ability was discussed by using the walking index of spinal cord injuryⅡ (WISCⅠ Ⅱ) and 10 minutes’ walking time (10 MWT). Before and after treatment, functions of daily life were evaluated by using Barthel index (MBI). Bladder function was also discussed, and levels of brain-derived nerve growth factor in peripheral blood (BDNF), nerve growth factor (NGF), astrogenic calcium binding protein S-100 β (S-100 β), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were also detected. Result:After the treatment, scores of sensory and motor in ASIA scale in observation group were all higher than those in control group (P<0.01). Score of WISCⅠ Ⅱ in observation group was higher than that in control group, and 10 MWT was shorter than that in control group (P<0.01). Times of leakage of urine, urinary incontinence volume and residual urine volume were all less than those in control group, and bladder volume was more than that in control group (P<0.01). Functions of daily life in observation group were better than those in control group (Z=1.967, P<0.05), and the levels of BDNF and NGF were higher than those in control group, while levels of S-100 β, TNF-α and IL-1β were lower than those in control group (P<0.01). Conclusion:On the basis of routine comprehensive rehabilitation therapy of western medicine, Jiajian Fuyuan Huoxuetang combined with electroacupuncture can promote the recovery of nervous function, improve sensory and motor function, improve bladder function and daily living ability, and promote the expression of neurotrophic factors and inhibit inflammatory reaction in neurological rehabilitation for the patients with spinal cord injury.

Analysis of influencing factors and countermeasures for adverse reactions after retinal laser photocoagulation / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the influencing factors and coping strategies of adverse reaction after retinal laser photocoagulation.<p>METHODS: Retrospective analysis of 1 165 cases of adverse reactions in patients with retinal laser photocoagulation in our hospital.<p>RESULTS: The present study included 50 patients with adverse reactions in 1 165 patients(4.29%), including 13 patients with nausea and vomiting(1.12%), 21 patients with dizziness, chest tightness, cold sweat(1.80%), 4 cases with yawning and drowsiness(0.34%), 12 cases with syncope(1.03%), and no death. There was no difference in the incidence of adverse reactions between men and women(3.68% <i>vs</i> 5.14%, χ2=1.474, <i>P</i>=0.225); there was a difference in the incidence of adverse reactions among patients of different ages(χ2=48.817, <i>P</i><0.05)and under 40. The incidence of adverse reactions was higher than that of patients aged 40 and older, and the incidence of adverse reactions in men under 40 was significantly lower than that in women(8.46% <i>vs</i> 18.60%, χ2=6.094, <i>P</i>=0.014). The adverse reactions of patients with different fundus diseases were different.<p>CONCLUSION: The incidence and degree of postoperative adverse reactions in patients with fundus diseases treated by retinal laser photocoagulation are different, so we should understand the history and mental state of the patients in detail before operation, and prepare the patients with high risk factors and possible serious reactions in time to ensure the safety of the patients.

Septic shock caused by Rhizobium radiobacter in an elderly woman: A case report.

RATIONALE: Rhizobium radiobacter is a Gram-negative pathogen present in soil and plants. Cases of R radiobacter infection in immunocompromised hosts have been sporadically reported. However, septic shock caused by R radiobacter is rarely seen. PATIENT CONCERNS: Here, we describe an elderly patient with a rapid progression of watery diarrhea, anorexia, fever, weakness, oliguria, and shock. Blood results showed increased total white blood cell count and C-reactive protein. Arterial blood gas results showed hypoxia and elevated lactate level. The Sequential Organ Failure Assessment score was 11. Blood culture at admission showed Gram-negative bacteria, which were later confirmed as R radiobacter. DIAGNOSIS: Septic shock caused by R Radiobacter. INTERVENTIONS: The patient was treated with intravenous cefoperazone/sulbactam and sequential oral levofloxacin. OUTCOMES: The patient recovered completely. CONCLUSION: R radiobacter may be considered as a potential opportunistic pathogen that may cause severe sepsis in elderly patients, especially those with multiple underlying diseases.

Advanced research and technology development of antibody-drug conjugates / 药学学报

Although numbers of naked antibodies showing clinical efficacy as single agents, their therapeutic effect is limited. Chemotherapy is very effective but with relatively large side effects, so conjugation of small chemotherapeutic drugs to antibodies is one of the important methods to enhance therapeutic potential of antibodies. Antibody-drug conjugates (ADCs) represent a promising therapeutic approach for cancer patients by combining the antigen-targeting specificity of monoclonal antibodies (mAbs) with the cytotoxic potency of chemotherapeutic drugs. These modified antibodies are expected to selectively deliver chemotherapeutic drugs to tumor cells and provide sustained clinical benefit to cancer patients, at the same time, minimizing systemic toxicity. ADCs are expected to bring together the benefits of highly potent drugs on the one hand and selective binders of specific tumor antigens on the other hand. However, designing an ADC is very complex, requiring thoughtful combination of antibody, linker, and payload drugs in the context of a target and a defined cancer indication. Although many challenges remain, recent clinical success has generated intense interest in this therapeutic class.

Simultaneous Determination of Seven Isosteroidal Alkaloids in Fritillaria wabuensis by HPLC-ELSD Method / 中国药学杂志

OBJECTIVE: To establish an effective high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) method to simultaneously detect multi alkaloids in Fritillaria wabuensis, which can be used to control the quality of Fritillaria wabuensis. METHODS: The method was conducted on an InertSustain C18 column (4.6 mm×250 mm, 5 μm) with 0.1% TFA (V/V) aqueous solution-acetonitrile as mobile phase eluted in a gradient program. RESULTS: The method had good linearity (r2≥0.999 0).The accuracy was between 90% and 105%. This method was simple, sensitive, accurate and specific, which can be applied to determine imperialine-3-β-D-glucoside, imperialine, peimisine, verticine, verticinone, isoverticine and chuanbeinone in Fritillaria wabuensis. CONCLUSION: Nearly all fritillariae alkaloids can be detected under this condition, therefore, the chromatogram can be used as the characteristic chromatogram to testify different kinds of Fritillaria.The method can also be combined with mass spectrometric detector. It′s the first time for isoverticine and chuanbeinone to be detected in Fritillaria wabuensis by HPLC and also the first time for seven isosteroidal alkaloids in Fritillaria wabuensis to be simultaneously determined.

[miR-362 regulates the proliferation and apoptosis of porcine immature Sertoli cells through targeting the ZNF644 gene].

In testicular tissue, immature Sertoli cell proliferation ability determines the size of mature Sertoli cell populations, which further regulates the spermatogenesis in the adult male animals. Studies have demonstrated that microRNAs (miRNAs) participate in the regulation of the immature Sertoli cell proliferation and apoptosis, but the functions of most identified miRNAs remain unclear. In this study, based on previous RNA-seq results, we analyzed the regulatory role (s) of miR-362 in proliferation and apoptosis of porcine immature Sertoli cells. The ZFN644 gene was predicted to be a target gene of miR-362 using bioinformatics methods. The expression levels of miR-362 and ZNF644 gene were measured using qRT-PCR assay in developing porcine testicular tissues and in immature Sertoli cells transfected with either miR-362 mimic or miR-362 inhibitor. The dual luciferase reporter assay was used to determine the regulatory relationship between miR-362 and ZNF644. The results showed that a putative target site of miR-362 was located in the 3'UTR of ZNF644. The expression of miR-362 was significantly and negatively correlated with ZNF644 expression in the developing porcine testicular tissues. Co-transfection of miR-362 and psiCHECK2-ZNF644-WT 3'UTR luciferase vector significantly suppressed luciferase activity. The ZNF644 gene expression level was significantly regulated by miR-362, demonstrating that miR-362 targets ZNF644 gene and inhibits its expression in porcine immature Sertoli cells. Flow cytometry, CCK8, and EdU assays were used to measure the effects of over-expression of miR-362 or knockdown of ZNF644 on porcine immature Sertoli cell proliferation; Annexin V-FITC/PI staining assays and qRT-PCR technology were used to test the apoptosis and the expression levels of cell survival-related genes, respectively. Over-expression of miR-362 and knockdown of ZNF644 arrested the porcine immature Sertoli cells in G1 and G2 phases of the cell cycle, respectively, and inhibited proliferation, enhanced apoptosis in the porcine immature Sertoli cells, and significantly regulated the expression levels of cell survival-related genes. Taken together, these data indicate that miR-362 inhibits proliferation and promotes apoptosis in porcine immature Sertoli cells by targeting the ZNF644 gene, thereby providing the scientific basis for further study on the function(s) of miR-362 in the porcine spermatogenesis.

[Variations in growth response and cadmium accumulation of different gender Morus alba seedlings to combined treatments of Cd and acid rain]. / ä¸åŒæ€§åˆ«æ¡‘æ ‘å¹¼è‹—å¯¹é•‰ä¸Žé ¸é›¨å¤åˆå¤„ç†çš„ç”Ÿé•¿å“åº”åŠé•‰ç§¯ç´¯å·®å¼‚.

To investigate sex-specific adaptive responses and cadmium accumulation of Morus alba seedlings, we analyzed growth parameters, photosynthetic capacity and chlorophyll fluorescence parameters, and Cd accumulation and allocation of Qiangsang 1 (female) and Nongsang 14 (male) under different treatments [cadmium (Cd, 100 mg·kg-1), acid rain (AR, pH 3.0) and their combinations (Cd+AR)]. When exposed to Cd stress alone, females showed higher cadmium content in root, stem and leaf than males. The root, stem and total biomass, and maximum net photosynthetic rate (Amax) of males were significantly decreased, while those of females (except Amax) showed no significant changes. Cd had no effect on maximal photochemical efficiency (Fv/Fm), photochemical quenching coefficient (qP), non-photochemical quenching coefficient (qN) in both sexes. When exposed to Cd+AR stress, the total biomass and Amax of males and females decreased. Compared to Cd stress alone, the cadmium content in root and leaf of females significantly increased while those of males did not under Cd+AR. In addition, Cd+AR stress had no effect on Fv/Fm and qP, but significantly increased qN in both sexes. Our results suggested that females had greater tolerance than males when exposed to Cd stress alone in short term. Acid rain would decrease the tolerance of females to Cd stress, which might attribute to the enhanced absorption and accumulation of cadmium.