[Clinical feature changes of a COVID-19 patient from mild to critical condition and cardiopulmonary pathological results].

Objective: To analyse the clinical history, laboratory tests and pathological data of a patient who suffered from novel coronavirus pneumonia(COVID-19) and provide reference for the clinical treatment of similar cases. Methods: Data of clinical manifestation, laboratory examination, bronchoscopy, echocardiography and cardiopulmonary pathological results were retrospectively reviewed in a case of COVID-19 with rapid exacerbation from mild to critical condition. Results: This patient hospitalized at day 9 post 2019 novel coronavirus(2019-nCoV) infection, experienced progressive deterioration from mild to severe at day 12, severe to critical at day 18 and underwent extracorporeal membrane oxygenation(ECMO) and continuous renal replacement therapy(CRRT) as well as heart lung transplantation during day 28-45 post infection, and died at the second day post heart and lung transplantation. The patient had suffered from hypertension for 8 years. At the early stage of the disease, his symptoms were mild and the inflammatory indices increased and the lymphocyte count decreased continuously. The patient's condition exacerbated rapidly with multi-organ infections, and eventually developed pulmonary hemorrhage and consolidation, pulmonary hypertension, right heart failure, malignant ventricular arrhythmias, liver dysfunction, etc. His clinical manifestations could not be improved despite viral RNAs test results became negative. The patient underwent lung and heart transplantation and finally died of multi organ failure at the second day post lung and heart transplantation. Pathological examination indicated massive mucus, dark red secretions and blood clots in bronchus. The pathological changes were mainly diffused pulmonary hemorrhagic injuries and necrosis, fibrosis, small vessel disease with cardiac edema and lymphocyte infiltration. Conclusions: The clinical course of severe COVID-19 can exacerbate rapidly from mild to critical with lung, liver and heart injuries.

[Clinical analysis for preset double J tube in percutaneous nephrolithotripsy].

Objective: To evaluate the clinical application and effect of preseting Double-J ureteric stent in percutaneous nephrolithotripsy. Method: 74 cases of renal calculi treated with PCNL in our hospital during June 2014 to February 2017 were retrospectively analyzed. Of 74 cases, 54 was male, 20 was female. All cases were aged 27 to 78, the mean age was (49.5±12.3) years old. The diameter of the stone was 20 to 59 mm, and the mean diameter was of (29.4±4.3) mm.Our Surgical methods was first putingFr6 double J tube in abnormal ureteral in advance in lithotomy position, then indwellingthree-way Foley catheter and clipping drainage port, perfusingirrigation port with 3 000 ml saline from 60-80 cm height.Perfusingsaline through irrigation port in prone position, we produce artificial hydronephrosis, then indwelling channel Fr20 through B ultrasound guided percutaneous nephrostomy, and removing renal calculi using holmium laser lithotripsy. Results: All patients were successfully completed percutaneous nephrostomy and indwell Fr20 channel, mean channel set up time (8.0 ±2.0) min, mean operation time (79±46) min, mean decline of hemoglobin (17.0±4.0) g/L, mean serum creatinine increased(3.1±1.1) µmol/L, one-stage stone-free rates 81.1%, complication rate 8.1% (1 case injured pleura and suffered from pneumothorax, 1 case suffered from massive hemorrhage of renal arteriovenous fistula after operation, 4 cases suffered postoperative fever). Conclusion: Advance in percutaneous nephrolithotripsy indwelling double J tube is a safe and feasible method, which is advantageous to the percutaneous renal puncture and the establishment of channels, and can avoid the blindness of along the line of indwelling double J.

Overexpression of clusterin correlates with tumor progression, metastasis in gastric cancer: a study on tissue microarrays.

Clusterin (CLU) is expressed in a wide variety of human tissues and fluids. Overexpression of cytoplasmic clusterin (sCLU) has been implicated in cancer development and progression. The aim of the present study is to evaluate the association of sCLU overexpression with clinicopathological features of human gastric carcinomas (GC).We constructed a gastric cancer tissue microarray containing 173 primary gastric carcinomas and 70 paired non-neoplastic mucosa specimens. The expression of sCLU was studied by immunohistochemistry. The correlations between sCLU expression and clinicopathological features, p53 abnormality, as well as Ki67 activation were analyzed. Overexpressions of sCLU was detected in 28.5% (n=165) of primary GCs by immunohistochemical staining, but not in non-neoplastic mucosa. Clinical association study found that overexpression of sCLU was significantly correlated with lymph-node metastasis (p < 0.001), tumor invasion (p < 0.001) and TNM stage (p < 0.001). In Kaplan-Meier survival analysis, overexpression of sCLU was significantly correlated with unfavorable survival in advanced GCs (p < 0.03). Furthermore, the association of sCLU with abnormal expression of p53 was ascertained. These results suggested that overexpression of sCLU was involved in the progression of GC and it's oncogenic function might be associated with p53 abnormality. Overexpression of sCLU seems to be related with patient's shorter survival in late stage GC.

Clusterin plays an important role in hepatocellular carcinoma metastasis.

To identify genes associated with tumor metastasis in hepatocellular carcinoma (HCC), gene expression profiles between a pair of primary HCC (H2-P) and their matched metastatic HCC (H2-M) were compared. Overexpression of clusterin (CLU) was found in H2-M cells. To determine the roles CLU played in HCC metastasis, CLU was transfected into H2-P cells. Overexpression of CLU in H2-P cells increased cell migration by twofold in vitro and formation of metastatic tumor nodules in liver by eightfold in vivo. To evaluate the correlation of CLU expression with HCC metastasis, the expression levels of CLU in HCCs were investigated using a tissue microarray (TMA) containing 104 pairs of primary HCCs and their matched metastases. The frequency of CLU overexpression increased significantly in metastatic HCCs (59.1%) compared with that in primary tumors (32.6%, P<0.001). To gain additional insight into the function of CLU, the expression profile of H2P-CLU was compared with vector-transfected H2-P cells by cDNA microarray. A total of 35 upregulated and 14 downregulated genes were detected in H2P-CLU. One of the upregulated genes known as YKL-40, which is implicated in matrix-remodeling and metastasis, was further studied using TMA. A significant correlation (P<0.001) between the expression levels of YKL-40 and CLU was observed, implying that the CLU-YKL-40 pathway may play an important role in HCC metastasis.

miR-625 suppresses tumour migration and invasion by targeting IGF2BP1 in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most prevalent malignancies and the third leading cause of cancer-related deaths worldwide. Tumour metastasis is one of the major causes of high mortality. microRNAshave been implicated in HCC metastasis. In this study, we found that miR-625 was frequently downregulated in HCC samples. A decrease in miR-625 was significantly correlated with lymph node anddistance metastasis (P=0.013), the presence of portal venous invasion (P=0.036), tumor-node-metastasis (TNM) stage (P=0.027) and unfavourable overall survival (P=0.003). Compared with primary tumours, miR-625 expression was markedly reduced in portal venous metastatic tumours. Re-expression of miR-625 in HCC cells was remarkably effective in suppressing cell migration andinvasiveness in vitro and in vivo. Mechanistically, miR-625 was confirmed to downregulate IGF2 mRNA-binding protein 1(IGF2BP1) directly, the expression of which was inversely correlated with the level of miR-625 in HCC cell lines and tissues. High expression of IGF2BP1 was frequently found in HCC samples, and associated with poor prognosis. Knockdown of endogenous IGF2BP1 by siRNA exhibited similar effects as the overexpression of miR-625, whereas overexpression of IGF2BP1 (without the 3'-UTR) abrogated miR-625-mediated metastasis inhibition. Interference of the PTEN/HSP27 pathway contributed to miR-625-mediated metastasis inhibition. Taken together, our data suggest that miR-625 might function as an antimetastatic miRNA to have an important role in HCC progression by modulating the IGF2BP1/PTEN pathway. The newly identified miR-625/IGF2BP1 axis represents a new potential therapeutic target for HCC treatment.

Subcellular localization of merocyanine 540 (MC540) and induction of apoptosis in murine myeloid leukemia cells.

Subcellular localization of photosensitizers is thought to play a critical role in determining the mode of cell death after photodynamic treatment (PDT) of leukemia cells. Using confocal laser scanning microscopy and fluorescent organelle probes, we examined the subcellular localization of merocyanine 540 (MC540) in the murine myeloid leukemia M1 and WEHI 3B (JCS) cells. Two patterns of localization were observed: in JCS cells, MC540 was found to localize on the plasma membrane and mitochondria; and in M1 leukemia cells, MC540 was found to localize on lysosomes. The relationship between subcellular localization of MC540 and PDT-induced apoptosis was investigated. Apoptotic cell death, as judged by the formation of apoptotic nuclei, was observed 4 h after irradiation in both leukemia cell lines. Typical ladders of apoptotic DNA fragments were also detected by DNA gel electrophoresis in PDT-treated JCS and M1 cells. At the irradiation dose of 46 kJ/m2 (LD90 for JCS and LD86 for M1 cells), the percentage of apoptotic JCS and M1 cells was 78 and 38%, respectively. This study provided substantial evidence that MC540 localized differentially in the mitochondria, and the subsequent photodamage of the organelle played an important role in PDT-mediated apoptosis in myeloid leukemia cells.

A comparison of the photodynamic effects of temoporfin (mTHPC) and MC540 on leukemia cells: efficacy and apoptosis.

The photodynamic effects of temoporfin (meso-tetrahydroxyphenylchlorin, mTHPC) and merocyanine 540 (MC540) in murine myeloid leukemia M1 and WEHI 3B (JCS) cells were compared. The mTHPC was found to be more potent and selective. At a lethal dosage of 90% killing (LD90), only 1.3 microM of mTHPC and 4.2 kJ/m2 of light irradiation was required, which was a 20-fold lower drug concentration and 11-fold smaller light dose than that required when using MC540. Meanwhile, three times less, or 15%, of the coincubated erythrocytes were destroyed by mTHPC than by MC540. Confocal micrographs showed that both drugs accumulated diffusely inside the cytoplasm in a very similar fashion, but mTHPC induced a more extensive apoptosis in photosensitized JCS cells. For example, at LD90, mTHPC practically killed all JCS cells via apoptosis and cleaved the DNA to extremely small 150 base-pair fragments. In contrast, among the JCS cells killed by MC540, about 88% died via apoptosis and large DNA fragments were abundant. Relative to MC540, the ability of mTHPC to trigger large-scale and thorough apoptosis in leukemia cells may help explain its potency and selectivity.

[Comparison of point mutation induced by ethylnitrosourea in human fetus and Syrian hamster pulmonary epithelial cells].

Cytotoxicity and mutation at the HGPRT (hypoxanthineguanine phosphoribosyl transferase) locus induced by ethylnitrosourea (ENU) in human fetus and Syrian hamster pulmonary epithelial cells were studied. A dose-dependent relationship between ENU and both parameters was observed. Cell killing of ENU to both cell lines was similar when determined as a reduction in colony-forming efficiency. However, after treatment with 0.01 mg/ml of ENU, the frequency of point mutation was zero in the human cells, but 4.1/10(5) cells in the hamster cells. ENU at 0.8 mg/ml induced 248.4 mutants per 10(5) cells in the hamster cells, but only 9.4 mutants per 10(5) cells in the human cells. The average frequency of mutation from all doses in the hamster cells was 10.5-fold higher than in human cells. These results imply that hamster cells are more sensitive to mutation induction than human cells and the cytotoxic effect of ENU may not be responsible for point mutation in both cell lines. These results support our previous finding that human chromosomes are more stable than those of rodents.

[Pathology of primary non-Hodgkin's lymphoma of bone].

35 cases of primary non-Hodgkin's lymphoma (NHL) of bone are analysed. According to the diagnostic criteria of malignant lymphoma introduced at the National Conference on Lymphoma, they were divided into 3 groups: B cell derivative, T cell derivative and histiocytic derivative lymphomas. NHL of the bone was all of diffuse infiltration type, in which the incidence of T-cell and histiocytic derivative lymphomas was relatively frequent. X-ray changes and frequent complications of pathologic fracture in the tumor might be explained by the penetrating and osteolytic growth and direct morphologic osteolytic action of the tumor cells. But local fibrosis is more commonly found, on the basis of which reactive osseous hyperplasia is formed. Sometimes there are more macrophages and lymphocytes admixed around the tumor. It shows the immune reaction of the organism to the tumor.

Can anisodamine be a potential substitute for high-dose atropine in cases of organophosphate poisoning?

A case of organophosphate (OP) poisoning was admitted to the emergency room. The patient accepted treatment with pralidoxime (PAM), atropine, and supporting therapy. It was observed that even after 22 h after treatment, 960 mg of atropine was not enough for the patient to be atropinized. However, a 160-mg follow-up treatment of anisodamine was quite enough for atropinization after 4 h. As a case report, more studies are required before any definite conclusion can be reached regarding the use of anisodamine as a potential substitute for high-dose atropine in cases of OP poisoning.

Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma.

Clinically, human glioblastoma (GBM) may develop de novo or from a low-grade glioma (secondary GBM), and molecular alterations in the two pathways may differ. This study examined the status of Survivin expression and apoptosis in 30 primary and 26 secondary GBMs. Our results show that cytoplasmic Survivin positivity was significantly (P<0.001) more frequent in primary GBMs (83%) than that in secondary GBMs (46%). In addition, an inverse correlation of cytoplasmc Survivin positivity with GBM apoptotic index, and a positive association between cytoplasmic Survivin and size of the tumours were observed. These results suggest that cytoplasmic Survivin, via its antiapoptotic function, may be involved in the tumorigenesis of many primary GBMs, but only in a small fraction of secondary GBMs. Furthermore, the overall progression times from low-grade precursor lesions to secondary GBMs were significantly shorter (P<0.05) in cytoplasmic Survivin-positive cases (mean, 15.6 months) than those in Survivin-negative cases (mean, 23.8 moths), and the positive expression level of Survivin in cytoplasm was upregulated in most secondary GBMs when compared to matched pre-existing low-graded lesions. These results suggest that the increased accumulation of Survivin in the cytoplasm of more malignant glioma cells may prove to be a selective advantage, thus accelerating progression to a more aggressive phenotype.

[Ectopic osteogenesis of bone marrow stromal cells induced by bone morphogenetic protein].

OBJECTIVE: To investigate the ectopic osteogenesis of bone marrow stromal cells (MSC) induced by bone morphogenetic protein(BMP) in vitro and in vivo, providing the experimental evidence for making an artificial bone with its own capacity of bone formation. METHODS: MSC were separated and cultured from bone marrow of Wistar rats, MSC were co-cultured with BMP in vitro (cultured in plate and diffuse chamber). Artificial coral hydroxyapatites (CHA) with MSC and BMP were implanted into dorsal muscles of Wistar rats, their bone formation were observed by morphological examination, histochemistry and immunohistochemistry. RESULTS: Only cartilaginous matrix were produced by MSC in vitro (cultured in plate and diffuse chamber), and both cartilaginous and bone matrix production within the combined grafts were seen. The bone formation of experimental groups (CHA + BMP + MSC) was stronger than that of control A(CHA + MSC) and control B(CHA). CONCLUSION: It may be possible to produce an artificial bone with its own capacity of bone formation by combined graft (CHA + BMP + MSC). There may be multiple factors as well as BMP inducing bone formation both in the whole body and the location of the implantation. Further research on these factors will have the significance for making the ideal artificial bone.

A non-isotopic method for the detection of telomerase activity in tumour tissues: TRAP-silver staining assay.

Telomerase activity has been observed in a high proportion of specimens from a wide variety of human malignant tumours, but not in non-neoplastic tissues. Measurement of telomerase activity is a possible diagnostic maker for malignant tumours. A non-isotopic telomeric repeat amplification protocol (TRAP) silver staining method for the detection of telomerase activity in human bone tumour tissues is described. The TRAP-silver staining method was quick, safe, and effective, and can be used as a routine diagnostic method for the detection of telomerase activity in bone tumours.

The expression of ADAM12 (meltrin alpha) in human giant cell tumours of bone.

AIMS: To examine the expression of ADAM12 (meltrin alpha), a member of the disintegrin and metalloprotease (ADAM) family, in human giant cell tumours of the bone, skeletal muscle tissue from human embryos, and human adult skeletal muscle tissue. METHODS: ADAM12 mRNA was detected by reverse transcription polymerase chain reaction and in situ hybridisation. RESULTS: ADAM12 mRNA was detected in 14 of the 20 giant cell tumours of bone and in three of the six tumour cell cultures. The expression of ADAM12 in cells cultured from the tumour was linked to the presence of multinucleated giant cells. ADAM12 mRNA could not be detected in the five adult skeletal muscle tissue samples, although it was found in the two embryonic skeletal muscle tissue samples. ADAM12 mRNA was localised to the cytoplasm of multinucleated giant cells and some mononuclear stromal cells. CONCLUSIONS: These results indicate that multinucleated giant cells are formed by the cell fusion of mononuclear stromal cells in giant cell tumours of bone and that ADAM12 is involved in the cell fusion process.