[Clinical features and prognostic analysis of transarterial chemoembolization combined with targeted immunotherapy in the treatment of patients with hepatitis B virus-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis].

Objective: To study the clinical features and prognostic impact of transarterial chemoembolization (TACE), immune checkpoint inhibitors (ICIs), and tyrosine kinase inhibitors (TKIs) combination therapy regimens in the treatment of patients with hepatitis B virus-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis. Methods: Patients with HBV-related intermediate-and advanced-stage hepatocellular carcinoma (HBV) who visited the Affiliated Hospital of Xuzhou Medical University between January 1, 2020, and December 31, 2022, were enrolled. TACE+TKIs +ICIs combination therapy was used to treat all patients. The occurrence and factors influencing cholestasis, as well as the impact on prognosis after combined therapy, were analyzed. The measurement data were compared using a t-test and a non-parametric rank sum test. The count data was compared using the χ(2) test. The survival rates were compared using a log-rank test between different groups. Results: A total of 106 cases with HBV-related intermediate-and advanced-stage hepatocellular carcinoma were enrolled. The probabilities of secondary cholestasis within 3 and 6 months, 1, 2, and 3 years after TACE+ICIs+TKIs combination therapy were 9.4%, 12.3%, 14.2%, 24.5%, and 24.5%, respectively. Patients with secondary cholestasis had persistent symptoms and rapid progression. During the treatment course, the median survival time was significantly longer in patients with hepatocellular carcinoma without secondary cholestasis than that of patients with cholestasis (26.9 months vs. 13.7 months, respectively, P < 0.05). Secondary cholestasis, baseline aspartate aminotransferase, and prothrombin activity levels were independent risk factors that affected the survival and prognosis of patients treated with combination therapy. There was no statistically significant difference in the occurrence of other adverse reactions between the two groups with secondary and non-secondary cholestasis during the treatment course (47.5% vs. 43.3%, χ(2)=0.058, P = 0.810). Conclusion: TACE+ICIs+TKIs therapy combination is relatively common in the treatment of patients with HBV-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis. Moreover, accelerated disease progression is an independent risk factor affecting the survival and prognosis of patients.

[Analysis of CT features of 15 children with 2019 novel coronavirus infection].

Objective: To explore imaging characteristics of children with 2019 novel coronavirus (2019-nCoV) infection. Methods: A retrospective analysis was performed on clinical data and chest CT images of 15 children diagnosed with 2019-nCoV infection. They were admitted to the Third People's Hospital of Shenzhen from January 16 to February 6, 2020. The distribution and morphology of pulmonary lesions on chest CT images were analyzed. Results: Among the 15 children, 5 were males and 10 females, aged from 4 to 14 years. Five of the 15 children were febrile and 10 were asymptomatic on the first visit. The first nasal or pharyngeal swab samples in all the 15 cases were positive for 2019-nCoV nucleic acid. For their first chest CT images, 6 patients had no lesions, while 9 patients had pulmonary inflammatory lesions. Seven cases had small nodular ground glass opacities and 2 cases had speckled ground glass opacities. After 3 to 5 days of treatment, 2019-nCoV nucleic acid in a second respiratory sample turned negative in 6 cases. Among them, chest CT images showed less lesions in 2 cases, no lesion in 3 cases, and no improvement in 1 case. The remaining 9 cases were still positive in a second nucleic acid test. Six patients showed similar chest CT inflammation, while 3 patients had new lesions, which were all small nodular ground glass opacities. Conclusions: The early chest CT images of children with 2019-nCoV infection are mostly small nodular ground glass opacities. The clinical symptoms of children with 2019-nCoV infection are nonspecific. Dynamic reexamination of chest CT and nucleic acid are important.

[Analysis of CT features of 15 Children with 2019 novel coronavirus infection].

Objective: To explore imaging characteristics of children with 2019 novel coronavirus (2019-nCoV) infection. Methods: A retrospective analysis was performed on clinical data and chest CT images of 15 children diagnosed with 2019-nCoV. They were admitted to the third people's Hospital of Shenzhen from January 16 to February 6, 2020. The distribution and morphology of pulmonary lesions on chest CT images were analyzed. Results: Among the 15 children, there were 5 males and 10 females, aged from 4 to 14 years old. Five of the 15 children were febrile and 10 were asymptomatic on first visit. The first nasal or pharyngeal swab samples in all the 15 cases were positive for 2019-nCoV nucleic acid. For their first chest CT images, 6 patients had no lesions, while 9 patients had pulmonary inflammation lesions. Seven cases of small nodular ground glass opacities and 2 cases of speckled ground glass opacities were found. After 3 to 5 days of treatment, 2019-nCoV nucleic acid in a second respiratory sample turned negative in 6 cases. Among them, chest CT images showed less lesions in 2 cases, no lesion in 3 cases, and no improvement in 1 case. Other 9 cases were still positive in a second nucleic acid test. Six patients showed similar chest CT inflammation, while 3 patients had new lesions, which were all small nodular ground glass opacities. Conclusions: The early chest CT images of children with 2019-nCoV infection are mostly small nodular ground glass opacities. The clinical symptoms of children with 2019-nCoV infection are nonspecific. Dynamic reexamination of chest CT and nucleic acid are important.

Spirometra erinaceieuropaei severely infect frogs and snakes from food markets in Guangdong, China: implications a highly risk for zoonotic sparganosis.

Sparganosis is a parasitic disease caused by plerocercoid larvae of the genus Spirometra. In China, the main source of sparganosis is from Guangdong, 16.1% of the country's human sparganosis cases occur in this province. Frequent international trade of amphibians and reptiles in Guangdong may introduce new species of Spirometra into the local market. In this study, a large-scale, high-intensity sampling survey was conducted to find out the causative species and epidemic situation of Sparganosis in Guangdong. The prevalence of sparganum infection in five species of frogs (Boulengerana guentheri, Fejervarya multistriata, Hoplobatrachus chinensis, Pelophylax nigromaculatus and Quasipaa spinosa) and nine species of snakes (Elaphe carinata, Lycodon rufozonatum, Hypsiscopus plumbea, Ptyas dhumnades, P. korros, P. mucosa, Naja atra, Sinonatrix annularis and Xenochrophis piscator) was investigated in Guangdong, Southern China from May 2014 to August 2015. The results showed that 9.8% (50/511) of the frogs and 40.8% (141/ 346) of snakes were found to be infected by plerocercoids (spargana). To identify the species of the collected spargana, a partial sequence of the mitochondrial cytochrome c oxidase subunit1 gene (cox1) was amplified and sequenced. Phylogenetic analysis identified all the spargana specimens as Spirometra erinaceieuropaei. Our study indicated that S. erinaceieuropaei, a highly pathogenic parasite, is the only causative agent of sparganosis in Guangdong, China. This study suggests that the large numbers of frogs and snakes in food markets in Guangdong may impact public health in China by transmitting S. erinaceieuropaei sparganum. Additional steps should be considered by the governments and public health agencies to prevent the risk of food-associated Spirometra infections in humans in China.

EB-virus latent membrane protein 1 potentiates the stemness of nasopharyngeal carcinoma via preferential activation of PI3K/AKT pathway by a positive feedback loop.

Our previous study reported that Epstein-Barr virus(EBV)-encoded latent membrane protein 1 (LMP1) could induce development of CD44(+/High) stem-like cells in nasopharyngeal carcinoma (NPC). However, the molecular mechanisms that underlie modulation of cancer stem cells (CSCs) in NPC remain unclear. Here, we show that LMP1 induced CSC-like properties through promotion of the expression of epithelial-mesenchymal transition-like cellular markers and through alterations in differentiation markers. Furthermore, LMP1 activated and triggered phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway, which subsequently stimulated expression of CSC markers, development of side population and tumor sphere formation. This suggests that PI3K/AKT pathway has an important role in the induction and maintenance of CSC properties in NPC. Similarly, PI3K/AKT pathway was also activated by phosphorylase in LMP1-induced CD44(+/High) cells. In addition, LMP1 greatly increased expression of miR-21 and downregulated expression of the miR-21 target, PTEN. Overexpression of miR-21 by transfection of miR-21 mimics into LMP1-transformed cells led to phosphorylase-mediated activation of the PI3K/AKT pathway and induction of CSCs. On the contrary, phosphorylation of the PI3K/AKT pathway and the expression of CSC were reversed by an miR-21 inhibitor. The specific inhibitor (Ly294002) of PI3K/AKT pathway significantly decreased expression of miR-21 and CSC markers and upregulated the expression of PTEN, which indicates that miR-21 and PTEN are the downstream effectors of PI3K/AKT and that expression of these two effectors are related to the development of NPC CSCs. Taken together, our novel findings indicate that LMP1, PI3K/AKT, miR-21 and PTEN constitute a positive feedback loop and have a key role in LMP1-induced CSCs in NPC.

Genomewide analysis of intronic microRNAs in rice and Arabidopsis.

MicroRNAs (miRNAs) are potent regulators of gene transcription and posttranscriptional processes. The majority of miRNAs are localized within intronic regions of protein-coding genes (host genes) and have diverse functions in regulating important cellular processes in animals. To date, few plant intronic miRNAs have been studied functionally. Here we report a comprehensive computational analysis to characterize intronic miRNAs in rice and Arabidopsis. RT-PCR analysis confirmed that the identified intronic miRNAs were derived from the real introns of host genes. Interestingly, 13 intronic miRNAs in rice and two in Arabidopsis were located within seven clusters, of which four polycistronic clusters contain miRNAs derived from different families, suggesting that these clustered intronic miRNAs might be involved in extremely complex regulation in rice. Length analysis of miRNA-carrying introns, promoter prediction and qRT-PCR analysis results indicated that intronic miRNAs are coexpressed with their host genes. Expression pattern analysis demonstrated that host genes had a very broad expression spectrum in different stages of development, suggesting the intronic miRNAs might play an important role in plant development. This comparative genomics analysis of intronic miRNAs in rice and Arabidopsis provides new insight into the functions and regulatory mechanisms of intronic miRNAs in monocots and dicots.

Cadmium toxicity is alleviated by AtLCD and AtDCD in Escherichia coli.

AIMS: Arabidopsis thaliana l- and d-cysteine desulfhydrases (AtLCD and AtDCD) are two important H(2) S-generating enzymes. This study determined the effects of H(2) S derived from AtLCD and AtDCD on cadmium (Cd) toxicity in Escherichia coli. METHODS AND RESULTS: AtLCD and AtDCD were cloned into pET28a vectors and transformed into wild-type E. coli strain BL21(DE3), named BL21(LCD) and BL21(DCD). In the induced BL21(LCD) and BL21(DCD) compared with wild type, significantly higher H(2) S generation rates were observed. Additionally, higher survival rates, reduced contents of malondialdehyde (MDA) and hydrogen peroxide (H(2) O(2)), decreased activities of superoxide dismutase and catalase under 220 µmol l(-1) Cd stress were noted. We obtained similar results in the wild type treated with NaHS, a H(2) S donor. The above changes were substantially counteracted by the mixture of ammonia and pyruvic acid potassium (NH(3) + C(3) H(3) KO(3)), a synthetic inhibitor of H(2) S. CONCLUSIONS: AtLCD and AtDCD catalyse the H(2) S production, generating an ameliorating effect against Cd-induced oxidative stress and resulting in E. coli resistance to Cd toxicity. SIGNIFICANCE AND IMPACT OF THE STUDY: H(2) S as a gasotransmitter is certified to have an ameliorating effect against Cd toxicity, thus providing information for further research regarding the role of H(2) S in regulating resistance to the heavy metal stress in organisms.

Progress in cell membrane chromatography.

Cell membrane chromatography (CMC) was first established by He et al. in 1996. A bioaffinity chromatography technique, CMC has since proven to be an important method for studying drug-receptor interactions and screening active compounds from medicinal herbs. This paper briefly reviews the characteristics of the cell membrane stationary phase (CMSP), the CMC analytical system, and its applications.

Cesium cholate: determination of X-ray crystal structure indicates participation of the ring hydroxyl groups in metal binding.

The crystal structure of cesium cholate, C(24)H(36)(OH)(3) COOCs has been determined with three-dimensional X-ray diffractometer data. It crystallized in the monoclinic space group P2(1) with unit-cell dimensions a = 11.543(5) A, b = 8.614(3) A, and c = 12.662(5) A, beta(deg) = 107.95(2), V = 1197.7 A(3) and Z = 2. The atomic parameters were refined to a final r = 0.0269 and R(omega) = 0.0280 for 2342 observed reflections. Each Cs(+) is coordinated to 7 oxygen atoms from 5 different cholate anions with Cs-O distances ranging from 2.957(4) A to 3.678(5) A. In this crystal, 5 cholates are coordinated with 1 Cs(+), and 5 Cs(+) are coordinated with 1 cholate anion. Carboxyl and all the 3 ring hydroxyl groups of cholate anion participate in binding to Cs(+) simultaneously, and there is no water molecule coordinated with the Cs(+). The pattern of successive rows arranged with polar (p) and non-polar (n) faces in apposition leads to the formation of a sandwich sheet structure with polar and non-polar channels. The Cs ions lie within the polar interior of the sandwich. The H-bond network is reorganized in forming cesium cholate from cholic acid. All the oxygen atoms in cholate anion are involved in H-bonding reciprocally or with water molecules to form an extensive 3-dimensional network of H-bonds. Compared with cholic acid and other similar type of steroids, the coordination structure and H-bonding of Cs cholate crystal are distinct.

NO mediated increase of Fos protein and NMDA1A R mRNA expression in rat spinal cord during morphine withdrawal.

AIM: To investigate the effects of nitric oxide (NO) on activation of the rat spinal cord neurons during naloxone-precipitated morphine withdrawal. METHODS: Fos immunocytochemistry, NADPH-d histochemistry, Fos/NADPH-d double-labeling, intrathecal injection, antisense oligonucleotides (AS-ONs) techniques, and RT-PCR were used. RESULTS: Acute administration of naloxone and chronic administration of morphine did not change the expression of Fos protein and NADPH-d positive neurons, and there was no expression of Fos/NADPH-d double-labeled neurons in the spinal cord of rats. Morphine withdrawal increased the expression of Fos protein, NADPH-d positive, and Fos/NADPH-d double-labeled neurons, and they were observed in all the laminae of the rat spinal cord. Intrathecal injection of nNOS antisense oligonucleotides (nNOS-AS) inhibited the increase of Fos protein and NMDA(1A)R mRNA expression in the rat spinal cord during morphine withdrawal and decreased the scores of morphine withdrawal symptoms. The effect of nNOS-AS was greater than that of eNOS-AS. There was no effect in nNOS sense oligonucleotides (nNOS-S) group. CONCLUSION: NO mediated the increase of Fos protein and NMDA1A R mRNA expression in the rat spinal cord during morphine withdrawal.

[Intrathecal injection of NOS antisense oligonucleotides inhibits the increase of NMDA1AR mRNA expression in the spinal cord and brainstem of morphine-withdrawal rats].

Methods of reverse transcription polymerase chain reaction (RT-PCR), intrathecal injection and antisense drugs were used to study the effects of nitric oxide (NO) on the scores of morphine-withdrawal syndrome and the expression of NMDA1AR mRNA in rat spinal cord and brainstem. Intrathecal injection of NOS antisense oligonucleotides (AS-ONs) significantly decreased the scores of morphine-withdrawal symptoms. The effect of nNOS AS-ONs was greater than that of eNOS AS-ONs. The expression of NMDA1AR mRNA in the spinal cord and brainstem increased in morphine-dependent rats and increased to a greater extent in morphine-withdrawal rats. Intrathecal injection of nNOS AS-ONs significantly inhibited the increased expression of NMDA1AR mRNA in the spinal cord and brainstem of morphine-withdrawal rats. Intrathecal injection of eNOS antisense oligonucleotides inhibited the expression of NMDA1AR mRNA in the spinal cord of morphine-withdrawal rats, but did not in the brainstem. It is suggested that NO mediates morphine-withdrawal reaction and participates in modulating the expression of NMDA1AR mRNA in morphine-withdrawal rats.

[NO involvement in mediation of spinal neuron sensitization in morphine-withdrawal rats].

The effects of nitric oxide (NO) on the activation of spinal cord neurons were studied using immunocytochemistry, intrathecal injection and antisense oligonucleotides (AS-ONs) techniques in morphine-withdrawal rats. Acute administration of naloxone and chronic administration of morphine changed neither the expression of Fos-LI and NADPH-d positive neurons nor the expression of Fos/NADPH-d double-labeled neurons in the spinal cord of rats. Fos-LI, NADPH-d positive and Fos/NADPH-d double-labeled neurons were increased significantly in number in morphine-withdrawal rats and they were observed in all the laminae of the spinal cord. Intrathecal injection of L-NA, nNOS antisense oligonucleotides significantly inhibited the expression of Fos-LI in the spinal cord and decreased the scores for morphine-withdrawal symptoms in morphine-withdrawal rats, but not in nNOS-S group. The results suggest that NO mediates the spinal neurons sensitization in morphine-withdrawal rats.

The first organo-templated cobalt phosphate with a zeolite topology.

An organic molecule containing cobalt phosphate (denoted CoPO-GIS) which is isostructural with the zeolite gismondine has been synthesized under solvothermal conditions by using [Co(en)3]Cl3 and phosphoric acid as the reactants and ethylene glycol as the solvent. CoPO-GIS ((H3NCH2CH2NH3)0.5.CoPO4) crystallizes in the monoclinic space group C2/c with a = 14.744(3) A, b = 8.850(3) A, c = 10.062(3) A, beta = 131.609(19) degrees, and Z = 8. The structure consists of a 3-D network of strictly alternating CoO4 and PO4 tetrahedra interconnected by oxygen bridges, and the charge-balancing diprotonated ethylenediamine cations are highly ordered in the cages of the CoPO4 framework. CoPO-GIS is the only amine-containing cobalt phosphate with a known zeolite topology.

Cardiac responses activated by nicotine in canine ganglial plexus between aorta and pulmonary artery.

In order to study the location and function of nicotine-sensitive neurons of cardiac ganglial plexuses, microdissections of collections of fat pads were carried out, and nicotine (100 micrograms) was injected into the ganglial plexus between aorta and pulmonary artery (A-PGP) in 30 anesthetized open-chest dogs. There were numerous ganglia or neurons in A-PGP. Either positive or negative inotropic and chronotropic responses were elicited following injections of nicotine into A-PGP. Control injections of 0.1 ml saline into A-PGP and injections of nicotine (100 or 200 micrograms) into right marginal ganglial plexus did not elicit any cardiac responses. After acute decentralization, nicotine (100 micrograms) was again injected into the same locus of A-PGP. Some positive and negative responses could still be induced, but their frequencies were reduced. These suggest that nicotine can directly activate the efferent parasympathetic and sympathetic neurons and indirectly activate them by stimulating the afferent neurons existing on the surface of dog heart.

[Comparison between the effects of (-)- and (+)-gossypol on protein kinase C and protein kinase A].

Protein kinase C(PKC) and protein kinase A(PKA) purified from rat liver were incubated with (-)- and (+)-gossypol. The activity of PKC was significantly inhibited in a concentration-dependent manner by both (-)-gossypol and (+)-gossypol. (-)-gossypol was found to inhibit type I PKA, whereas the inhibitory action of (+)-gossypol for type I PKA was less potent. Both (-)-gossypol and (+)-gossypol showed inhibition for type II PKA only at high concentration. These results suggest that administration of gossypol might impair cellular signal transduction.