[Construction of zebrafish models for screening intracranial hemorrhage associated genes].

Objective: To construct zebrafish models for the screening of intracranial hemorrhage (ICH) associated genes. Methods: ICH zebrafish models were constructed through morpholino oligonucleotides (MOs) technique and microinjection technique, and multiple verification was performed from macro and micro perspectives. First, the normal wild-type AB strain zebrafish injected with control MO was used as the control group, and AB zebrafish embryos microinjected with MOs of genes related to development of neural crest-derived cells (NCDCs) were used as the study group, such as col8a1 MO, tfap2α MO, msx1a MO, msx2 MO, and dkk1a MO. Preliminary verification of the model was conducted under a white-light optical microscope. Then, the model was verified by Tg (flk1: gfp; gata1: dsRed) double transgenic zebrafish, with vascular endothelial cells labeled by green fluorescent protein (GFP) and red blood cell labeled by fluorescent protein (dsRed), and thus the location of cerebral hemorrhage can be observed more clearly. Specifically, zebrafish embryos were microinjected with Control MO as the control group and those microinjected with col8a1 MO as the study group. Then the embryos were cultured until 48 hours post-fertilization to observe the leakage of red blood cells under the confocal laser scanning microscope. Finally, Tg (flk1: gfp) transgenic zebrafish was used to verify the model based on the blood-brain barrier (BBB). Through the leakage of dextran-rhodamine and DAPI dyes, the destruction of BBB and the occurrence of cerebral hemorrhage in zebrafish were further clarified, and quantitative statistics were carried out to verify the relationship between NCDCs development related genes and cerebral hemorrhage phenotype, which proved that the modeling was effective. Results: The zebrafish with col8a1, tfap2α, and msx1 mutations in the study group had apparent ICH compared with wildtype zebrafish, and the prevalence of ICH was 18.18% (52/286), 23.04% (62/251), and 35.94% (23/64), respectively. While, the zebrafish with msx2 and dkk1a mutations rarely had ICH, with the ICH prevalence of 1.03% (1/97) and 1.15% (1/87), respectively. The prevalence of red blood cells leakage in Tg (flk1:gfp; gata1:dsred) double transgenic zebrafish injected with Control Mo and col8a1 Mo was 0.37% (1/273) and 18.18% (52/286) (P<0.001). The number of DAPI positive nuclei of Tg (flk1: gfp) transgenic zebrafish injected with Control Mo and col8a1 Mo was 10.05±5.27 and 60.35±3.96 (P<0.001), and the fluorescent intensity of midbrain parenchymal induced by dextran-rhodamin leakage was 2.54±4.70 and 5.13±3.52 (P<0.001). Conclusion: This study successfully constructs the ICH zebrafish models, and ICH-related genes are screened out, such as col8a1, tfap2α, msx1, and so on.

[Effect of D-dimer on the prognosis of patients with aneurysmal subarachnoid hemorrhage based on propensity score matching].

Objective: To evaluate the effect of D-dimer on the prognosis of patients with aneurysmal subarachnoid hemorrhage (aSAH). Methods: A total of 1 658 patients who were first diagnosed with aSAH in West China Hospital of Sichuan University from December 2013 to June 2019 were retrospectively analyzed. All patients were divided into four groups according to the median and quartiles of D-dimer level, including 415 cases, 414 cases, 414 cases, and 415 cases in groups Q1, Q2, Q3, and Q4, respectively. Groups Q2, Q3, Q4, and group Q1 were matched by propensity score matching (PSM), and the correlation between D-dimer and each outcome was analyzed by logistic regression. Since there is no general clinical classification standard for D-dimer, this study attempted to reclassify patients into groups q1 (<0.55 mg/L, 94 cases), q2 (0.55-1.65 mg/L, 435 cases), q3 (1.65-5.50 mg/L, 650 cases) and q4 (>5.50 mg/L, 303 cases) based on 1, 3, 5, 10 times of the upper limit of the current clinical reference value. Results: The age of 1 658 aSAH patients were (57±12) years, including 1 068 males and 590 females. After PSM based on the median and quartiles of D-dimer level, there were 318 cases, 318 cases, 251 cases, and 229 cases in groups Q1, Q2, Q3, and Q4, respectively. Compared with group Q1 (<1.23 mg/L), the risk of in-hospital infection (OR=2.14, 95%CI: 1.47-3.11, P<0.001), pneumonia (OR=2.22, 95%CI: 1.51-3.28, P<0.001), urinary tract infection (OR=1.75, 95%CI: 1.12-2.75, P=0.014) and intracranial rebleeding (OR=3.59, 95%CI: 1.30-9.91, P=0.013) group Q4 (>4.95 mg/L) was higher. Likewise, the risk of adverse outcomes in group Q4 was also higher than that in group Q1, including unfavorable outcome at discharge (OR=2.12, 95%CI: 1.43-3.14, P<0.001), mortality during hospitalization (OR=3.03, 95%CI: 1.26-7.33, P=0.014), mortality within 90 days (OR=2.33, 95%CI:1.29-4.22, P=0.005), mortality within 180 days (OR=1.92, 95%CI: 1.12-3.29, P=0.018), mortality within 1 year (OR=2.07, 95%CI:1.23-3.47, P=0.006) and mortality during the longest follow-up period (OR=1.97, 95%CI:1.26-3.09, P=0.003). After secondary grouping and PSM based on current clinical reference values, there were 90 cases, 87 cases, 90 cases, and 43 cases, respectively in groups q1, q2, q3 and q4. The risk of nosocomial infection (OR=2.26, 95%CI: 1.14-4.45, P=0.019), blood-borne infection (OR=8.86, 95%CI:1.08-72.78, P=0.042), poor prognosis at discharge (OR=4.92, 95%CI: 2.18-11.07, P<0.001), death within 180 days (OR=3.39, 95%CI: 1.04-11.08, P=0.043), death within 1 year (OR=3.23, 95%CI: 1.10-9.49, P=0.033), and death within the longest follow-up period (OR=3.28, 95%CI: 1.34-8.01, P=0.009) was still higher in group q4 than that in group q1. Conclusion: aSAH patients with high D-dimer level have a higher risk of complications and mortality during hospitalization and worse clinical prognosis.

[Advances in the study of non-infectious liver diseases and precancerous lesions of hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) is a major disease with a high degree of malignancy, and poor prognosis, which seriously endangers human health. Chronic viral hepatitis (HBV, HCV)-cirrhosis-liver cancer patterns of pathogenesis has been widely accepted. However, the relationship between non-infectious liver disease and HCC is not completely clear; thereby how various non-infectious liver diseases develop through precancerous lesions has attracted widespread attention to HCC. A full understanding of these precancerous lesions is likely to provide new ideas and strategies for the prevention and treatment of non-infectious liver disease-related HCC.

[Chloroquine inhibits viability of renal carcinoma cells and enhances sunitinib-induced caspase-dependent apoptosis].

OBJECTIVE: To determine whether chloroquine (CQ), an often used inhibitor of late autophagy and autophagosome/lyosome fusion, can inhibit proliferation of renal carcinoma cells and investigate its effect on sunitinib (ST)-induced apoptosis. METHODS: Renal carcinoma cell line 786 O and ACHN had been used as cellular model and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay was carried out to detect the cell viability in response to CQ or ST treatment. Both transmission electron microscope and immunoblotting had been employed to observe apoptotic and autophagic process. To examine the involvement of autophagy in ST-dependent apoptosis, autophagy had been inhibited either chemically or genetically via utilizing autophagy inhibitor or specific small interference RNA (siRNA) targeted to either Ulk1 (unc-51-like kinase 1) or LC3 (microtubule associated protein 1 light chain 3 fusion protein), two essential autophagic proteins. RESULTS: Both ST and CQ induced cell viability loss, indicating that either of them could inhibit renal cancer cell proliferation. Clone formation experiments confirmed the aforementioned results. Furthermore, the combined ST with CQ synergistically promoted the loss of cell viability. By transmission electron microscopy and immunoblotting, we found that the ST induced both autophagy and caspase-dependent apoptosis. While 3-MA, an early autophagy inhibitor, reduced the ST-induced cleavage of poly (ADP-ribose) polymerase-1 (PARP-1), a substrate of caspase 3/7 and often used marker of caspase-dependent apoptosis, CQ promoted the ST-dependent PARP-1 cleavage, indicating that the early and late autophagy functioned differentially on the ST-activated apoptotic process. Moreover, the knock down of either Ulk1 or LC3 decreased the ST-caused apoptosis.Interestingly, we observed that rapamycin, a specific inhibitor of mTOR (mammalian target of rapamycin) and an inducer of autophagy, also showed to inhibit cell viability and increased the cleavage of PARP-1 in the ST-treated cells, suggesting that autophagy was likely to play a dual role in the regulation of the ST-induced apoptosis. CONCLUSION: ST activates both apoptotic and autophagic process in renal carcinoma cells. Although autophagy precedes the ST-induced apoptosis, however, early and late autophagy functions differentially on the apoptotic process induced by this compound. Additionally, ST can coordinate with the inducer of autophagy to inhibit the cell proliferation. Further research in this direction will let us illuminate to utilize CQ as a potential drug in the treatment of renal carcinoma.

[Ventilator-associated pneumonia among premature infants <34 weeks' gestational age in neonatal intensive care unit in China: a multicenter study].

Objective: To investigate the incidence and pathogen distribution of ventilator-associated pneumonia (VAP) among preterm infants admitted to level Ⅲ neonatal intensive care units (NICU) in China. Method: A prospective study was conducted in 25 level Ⅲ NICU, enrolling all preterm infants <34 weeks gestational age admitted to the participating NICU within the first 7 days of life from May 2015 to April 2016. Chi-square test, t test and Mann-Whitney U test were used for statistical analysis. Result: A total of 7 918 patients were enrolled, within whom 4 623(58.4%) were males. The birth weight was (1 639±415) g and the gestational age was (31.4±2.0) weeks; 4 654(58.8%) infants required non-invasive mechanical ventilation and 2 154(27.2%) required intubation. Of all the mechanically ventilated patients, VAP occurred in 95 patients. The overall VAP rate was 7.0 episodes per 1 000 ventilator days, varying from 0 to 34.4 episodes per 1 000 ventilator days in different centers. The incidence of VAP was 9.6 and 6.0 per 1 000 ventilator days in children's hospitals and maternity-infant hospitals respectively, without significant differences (t=1.002, P=0.327). Gram-negative bacilli (76 strains, 91.6%) were the primary VAP microorganisms, mainly Acinetobacter baumannii (24 strains, 28.9%), Klebsiella pneumonia (23 strains, 27.7%), and Pseudomonas aeruginosa (10 strains, 12.0%). Conclusion: The incidence of VAP in China is similar to that in developed counties, with substantial variability in different NICU settings. More efforts are needed to monitor and evaluate the preventable factors associated with VAP and conduct interventions that could effectively reduce the occurrence of VAP.

[Strategy of points, lines and layers in needle assisted laparoscope functional modified neck dissection through bilateral breast approach].

The incidence of papillary thyroid carcinoma is growing rapidly. Especially in young patients, the patients have high demand for better cosmetic effect and less psychological trauma. The application of laparoscope technique in papillary thyroid carcinoma surgery should meet the needs of patients. The development of needle assisted technique make it easy to access the neck potential fascial spaces, and provide the basic instrument for laparoscope modified neck dissection. The application of needle assisted techniques mainly include suture suspension and V type needle retractor, MiniLap, 3 mm laparoscopic apparatus. Indications of this procedure is very important, and it is necessary to make full use of the "critical points" , "mark lines" and "fascia layers" to precise positioning in the operation. Furthermore, the levels of dissection must be determined based on the pattern of cervical lymph node metastasis in differentiated thyroid carcinoma, that will make the surgical procedures more safety.

[Efficiency of FibroScan and FibroTouch in liver stiffness measurement and fat quantification: a comparative analysis].

Objective: To investigate the efficiency of FibroScan(FS)and FibroTouch(FT)in liver stiffness measurement(LSM)and fat quantification through a comparative analysis. Methods: The outpatients or hospitalized patients who underwent LSM and fat quantification using FS and FT were enrolled. The differences in success rate and detecting parameters between FS and FT were analyzed, as well as the correlation between FS and FT values. The t-test was used for comparison of normally distributed continuous data between groups, and a one-way analysis of variance or the Kruskal-Wallis test was used for comparison between multiple groups. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. Results: A total of 1621 patients were enrolled. The success rates of FT and FS were 100% and 94.96%, respectively, and the success rate of FS was influenced by sex, age, body mass index, and biochemical markers of liver function. FT has a significantly shorter duration of single detection and a significantly lower number of times of single detection than FS(duration of single detection: 190.21±38.78 s vs 220.89±68.36 s, P < 0.01; number of single detection times: 10.31±1.32 vs 11.81±3.76, P < 0.01), as well as a significantly lower ratio of interquartile range to median of fat quantification in the same patient(5.39%±4.81% vs 17.18%±14.07%, P < 0.01). The LSM and fat quantification of FS were significantly correlated with those of FT(r = 0.645 and 0.620, both Based on the duration and number of times of single detection, success rate, and stability of fat quantification, FT seems to have a better detection efficiency than FS. The detection values of FT and FS can be calculated with regression equations < 0.01). The equations of linear regression were LSM(FT)= 4.435+0.477×LSM(FS); CAP(FT)= 134.71+0.456×CAP(FS). Conclusion: Based on the duration and number of times of single detection, success rate, and stability of fat quantification, FT seems to have a better detection efficiency than FS. The detection values of FT and FS can be calculated with regression equations.

Identification of pathway-related modules in high-grade osteosarcoma based on topological centrality of network strategy.

OBJECTIVE: The objective of this paper is to identify pathway-related modules which are defined as in high-grade osteosarcoma based on topological centralities analysis of networks. MATERIALS AND METHODS: Co-expression network was constructed by weighted gene co-expression network analysis (WGCNA) based on differentially expressed genes (DEGs). Pathway enrichment analysis was conducted by Kyoto Encyclopedia of Genes and Genomes (KEGG) database to detect pathway enriched genes. Pathway-related modules of pathway enriched genes were mined from the co-expression network. Then topological centralities (degree, closeness, stress and betweenness centrality) analyses for co-expression network and sub-networks were performed to explore hub genes. Validation of hub genes was carried out utilizing reverse transcription-polymerase chain reaction (RT-PCR) assays. RESULTS: There were 129 nodes and 1229 edges in co-expression network. We obtained a total of 16 hub genes and 11 pathway-related modules. Module 17 (Bladder cancer module) was the most significant module, which comprising 9 of 16 hub genes and 6 pathway enriched genes, taking intersection elements (CAV1 and CCND1). RT-PCR results showed that both of CAV1 and CCND1 in high-grade osteosarcoma were significantly differentially expressed compared with normal controls. CONCLUSIONS: This work may contribute to understanding the molecular pathogenesis and provide potential biomarkers for detections and effective therapies of high-grade osteosarcoma.

Pathway-based network analysis of myeloma tumors: monoclonal gammopathy of unknown significance, smoldering multiple myeloma, and multiple myeloma.

Although many studies have been carried out on monoclonal gammopathy of unknown significances (MGUS), smoldering multiple myeloma (SMM), and multiple myeloma (MM), their classification and underlying pathogenesis are far from elucidated. To discover the relationships among MGUS, SMM, and MM at the transcriptome level, differentially expressed genes in MGUS, SMM, and MM were identified by the rank product method, and then co-expression networks were constructed by integrating the data. Finally, a pathway-network was constructed based on Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, and the relationships between the pathways were identified. The results indicated that there were 55, 78, and 138 pathways involved in the myeloma tumor developmental stages of MGUS, SMM, and MM, respectively. The biological processes identified therein were found to have a close relationship with the immune system. Processes and pathways related to the abnormal activity of DNA and RNA were also present in SMM and MM. Six common pathways were found in the whole process of myeloma tumor development. Nine pathways were shown to participate in the progression of MGUS to SMM, and prostate cancer was the sole pathway that was involved only in MGUS and MM. Pathway-network analysis might provide a new indicator for the developmental stage diagnosis of myeloma tumors.

Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles.

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

A double-blind comparative study of Chinese herbal medicine Jinlianqingre Effervescent Tablets in combination with conventional therapy for the treatment of uncomplicated hand, foot, and mouth disease.

Chinese herbal medicine Jinlianqingre Effervescent Tablets (JET) are the recommended control measure for uncomplicated hand, foot, and mouth disease (HFMD) by the Ministry of Health of China. However, high-quality evidence to support this recommendation is limited. A total of 288 patients ranging in age from 1 to 13 years were randomly assigned to JET in combination with conventional therapy (mainly including the reduction of temperature by applying physical cooling paste or warm bathing), or conventional therapy with placebo group for 7 days. The objective was to test the hypothesis that JET combination therapy is more effective than conventional therapy for uncomplicated HFMD. A randomized, double-blind, placebo-controlled trial was designed. Our study showed that, compared with conventional therapy, the median time to fever resolution was significantly shorter in the JET combination therapy (8 vs. 80 h; p < 0.0001); the risk of fever resolution increased in the JET combination therapy [hazard ratio, 19.8; 95% confidence interval (CI), 12.8 to 30.7]; the median healing time of rash or oral ulcer was significantly shorter in the JET combination therapy (14 vs. 74 h; p < 0.0001); and the median symptom score for skin or oral mucosa lesions improved more rapidly in the JET combination therapy during the follow-up period. The median duration of hospital stay was 6 days in the JET combination therapy and 7 days in the conventional therapy (p < 0.0001). No significant adverse events and complications were found in both groups. The addition of JET to conventional therapy reduced fever clearance time, healing time of skin or oral mucosa lesions, and duration of hospital stay in children with uncomplicated HFMD.

Neuronal apoptosis in the developing cerebellum.

The following study analysed apoptosis in proliferative cells and migrating neurons of the developing cerebellum. The external granular layer, Purkinje cell layer and internal granular layer in the developing mouse cerebellar cortex were analysed by active caspase-3 immunohistochemistry, Hoechst 33258 staining and Western blot analysis. Immunocytochemistry results indicated that the peak of apoptosis appeared at postnatal days P8, P5 and P9 in the external granular layer, Purkinje cell layer and internal granular layer, respectively. Subsequently, in each region, the rate of apoptosis decreased with increasing age. In contrast, Western blot results demonstrated the highest expression of activated caspase-3 in the cerebellum at P5, followed by a subsequent decline and disappearance of expression by P14. Activated caspase-8 was expressed maximally at P10, and subsequently disappeared by P30. The results of this study suggest that the key period of neuronal apoptosis in the cerebellar cortex is between P0 and P14, indicating that this developmental period could be susceptible to treatment for congenital neurodegenerative diseases.

A randomised controlled trial of low-dose misoprostol and dinoprostone vaginal pessaries for cervical priming.

OBJECTIVE: We studied the efficacy of 25-microg misoprostol pessaries as either single or double dose compared with a 3-mg dinoprostone pessary for cervical priming. DESIGN AND SETTING: A randomised controlled trial in Singapore. POPULATION: One hundred and seventy-one women with term pregnancies and modified Bishop scores (mBS) < or =6 from 2003 to 2004. METHOD: Patients were randomised to single misoprostol dose, double misoprostol dose or the current dinoprostone regimen. MAIN OUTCOME MEASURES: Primary outcome was number of women who achieved favourable mBS >6 or active labour by day 2. Secondary outcomes were time interval from insertion to delivery, cardiotocographic abnormalities, delivery and neonatal outcome. RESULTS: More women in the misoprostol double-dose group (96.6%) and dinoprostone group (93%) achieved the primary outcome compared with the single-dose group (77.8%) (P = 0.003 and P = 0.03, respectively). There was no difference in secondary outcomes. More multiparous women achieve primary outcome compared with nulliparous women (odds ratio 0.21, 95% confidence interval 0.06-0.77). CONCLUSION: Double-dose misoprostol 25 microg is as effective as dinoprostone 3 mg inserts for cervical priming; both are more efficacious than a single-dose misoprostol pessary. Parity prognosticates the success of induction.

A randomised crossover comparison of patient-controlled sedation and patient-maintained sedation using propofol.

This randomised, crossover study compared patient-controlled sedation using boluses of propofol and patient-maintained sedation using a target-controlled infusion of propofol. Twenty-three patients aged 18-35 years having surgical removal of bilateral third molar teeth under local anaesthesia during two separate visits were studied. In the majority of patients, both techniques provided moderate sedation, good operating conditions, stable physiological parameters and a high degree of patient satisfaction. Two patients became over-sedated during patient-controlled sedation. The time taken for titration to adequate sedation was longer with patient-maintained sedation than with patient-controlled sedation [mean (SD) = 8.6 (3.7) min vs. 5.7 (3.1) min, p < 0.005]. The mean overall propofol consumption was similar with both techniques. The majority of patients preferred patient-maintained sedation to patient-controlled sedation, p < 0.05.

The use of a genetically engineered herpes simplex virus (R7020) with ionizing radiation for experimental hepatoma.

The herpes simplex virus (HSV) recombinant virus R7020 is an attenuated virus designed as a candidate for immunization against both HSV-1 and HSV-2 infections. It was extensively tested in an experimental animal system and in a healthy human adult population without significant untoward effects. We report on the use of R7020 with ionizing radiation as an oncolytic agent for hepatomas. Two hepatoma cell lines were studied, Hep3B and Huh7. R7020 replicated to higher titers in Hep3B cells than in Huh7 cells. Tissue culture studies correlated with hepatoma xenograft responses to R7020. R7020 was more effective in mediating Hep3B tumor xenograft regression compared with Huh7. Ionizing radiation combined with R7020 also showed differential results in antitumor efficacy between the two cell lines in tumor xenografts. Ionizing radiation enhanced the replication of R7020 in Hep3B xenografts. Moreover, the combination of ionizing radiation and virus caused a greater regression of xenograft volume than either R7020 or radiation alone. Ionizing radiation had no effect on the replication of R7020 virus in Huh7 xenografts. These results indicate that a regimen involving infection with an appropriate herpesvirus such as R7020 in combination with ionizing radiation can be highly effective in eradicating certain tumor xenografts.

[A historical review and some comments on the nuclear transplantation in fish].

In this review article, a brief history and main results of the studies on the nuclear transplantation (cloning) in fish was introduced. Late Professor T. C. Tung (TONG Di-Zhou), a noted Chinese Experimental Embryologist and his research group at the Institute of Zoology, the Chinese Academy of Sciences (CAS) in Beijing, China initiated and established successful technology of nuclear transplantation in fish in 1963. During the past 36 years, most of the studies on fish cloning were performed mainly by Chinese groups except only one article was published by authors in former USSR (Gasaryan et al. 1979), and the other one was published by Japanese authors in Japan (Niwa et al. 1999). The primary goals of the studies of Tung's group were: (1) to study the inter-relationship between the nucleus and the cytoplasm in terms of the controlling effects determined by nucleus or cytoplasm or both during the ontogenesis, cell differentiation and phenotypic expression in a developing animal; and (2) to produce fish clones of commercial importance for agricultural purposes. The most successful results that have been obtained by Tung's group in collaboration with the investigators at various fisheries institutions in China were the production of nucleocytoplasmic adult hybrid fish between different varieties, species, genera and subfamilies that produced viable offspring. Furthermore, these nucleocytoplasmic fish hybrid revealed that while most phenotypic characteristics are controlled by the nucleus, a few are controlled by the cytoplasm or by both. In addition, the resulting nucleocytoplasmic fish hybrids also showed some better characteristics of economic importance such as faster growth rate, increasing of protein content and reducing of fat content in muscle, etc. So far, no such kinds of evidences are available either in amphibian or in mammals. Another most important result obtained by other Chinese groups showed that the uncultured or cultured adult somatic cell in fish which can support the nuclear transplanted eggs developing into adults. They were: (1) a 17 month old gold-fish obtained by transplanting an adult erythrocytes into an enucleated egg (WU et al. 1982); (2) a subcultured kidney cell nucleus of Crucian carp (Carassius auratus) can support the transplanted enucleated egg of same species developing into a three years old adult with female sexual characteristics (CHEN et al. 1986) and (3) a cultured liver cell nucleus from grass carp (Ctenopharyngoden idellus) can support a nuclear transplanted unfertilized egg of the same species developing into an adult fish at least of one year old when that paper was published. Some discussions and comments for evaluating the results obtained from previous studies and suggestions for further investigations in this research field are also provided. More detailed information could be found in the book--"Cloning in Fish-nucleocytoplasmic Hybrids" which was written by Shaoyi Yan in English and published by International Union of Biological Sciences and Educational and Cultural Press Ltd. Hong Kong in 1998.

Replication-competent, nonneuroinvasive genetically engineered herpes virus is highly effective in the treatment of therapy-resistant experimental human tumors.

A genetically engineered, nonneurotropic herpes simplex virus (R7020) with a proven safety profile in both animals and humans was found effective in the treatment of large xenotransplanted tumors arising from a radiation- and chemotherapy-resistant human epidermoid carcinoma and a hormone-refractory prostate adenocarcinoma. R7020 replicated to high titer and caused rapid regression of the human tumor xenografts. Tumor destruction was accelerated in animals given both R7020 and fractionated ionizing radiation. Tumors arising from cells surviving one treatment with R7020 were fully susceptible to a second dose of virus. We conclude R7020 is an effective antitumor agent for non-central nervous system tumor xenografts with an excellent safety profile.

Primary structure of the human platelet serotonin 5-HT2A receptor: identify with frontal cortex serotonin 5-HT2A receptor.

Previous radioligand binding studies have demonstrated human platelet serotonin2A (5-HT2A) receptor binding sites. Pharmacological similarities between platelet and frontal cortex 5-HT2A receptor binding parameters have been demonstrated. However, it is not clear whether the platelet 5-HT2A receptor primary structure is identical to that of the brain receptor. Three overlapping cDNAs were obtained to span completely the coding region of the 5-HT2A receptor. These clones were sequenced with external and internal primers. The nucleotide sequence of human platelet 5-HT2A cDNA was identical to that reported for the human frontal cortex 5-HT2A receptor, except for nucleotide 102 (T-->C), which has been reported to represent a normal DNA polymorphism that does not alter the amino acid sequence. This finding may have implications in the study of neuropsychiatric disorders for which altered platelet 5-HT2A receptor binding has been demonstrated.