Positive SARS-CoV-2 RNA recurs repeatedly in a case recovered from COVID-19: dynamic results from 108 days of follow-up.

The evidence of long-term clinical dynamic on Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) RNA re-positive case are less. We performed a 108 days follow-up on dynamic clinical presentations in a case, who hospitalized three times due to the positive recurrence of SARS-CoV-2 RNA after discharge, to understand the prognosis of the 2019-Coronavirus disease (COVID-19). In this case, positive SARS-CoV-2 recurred even after apparent recovery (normal CT imaging, no clinical symptoms, negative SARS-CoV-2 on stool sample and negative serum IgM test) from COVID-19, viral shedding duration lasted for 65 days, the time from symptom onset to disappearance was up to 95 days. Erythrocyte-associated indicators, liver function and serum lipid metabolism presented abnormal throughout during the observation period. Awareness of atypical presentations such as this one is important to prompt the improvement of the management of COVID-19.

[Detection of peripheral blood HBV-LHBs transactivation function and its relationship with anti-viral efficacy].

OBJECTIVE: Explore the serum of patients with CHB of HBV large envelope protein (HBV-LHBs) trans-activation function and antiviral therapy effect relationship. METHODS: 60 cases of anti-viral treatment of patients with chronic hepatitis B to take every 3 months HBVDNA, HBV-LHBs, as well as detection of hepatitis B immune markers to observe the changes in indexes. RESULTS: Income group 60 cases of anti-virus group HBVDNA with HBV-LHBs have a higher detection rate of the consistency of the results found no statistical significance (P > 0.05), HBV-LHBs-positive rate and positive rate of HBeAg differences (chi2 = 4.08, P < 0.05). After 24 months of antiviral therapy HBV-LHBs expression always HBVDNA in 29 cases of which occurred 24 months after the negative reaction of the 20 cases, continuous positive were seven cases of non-negative. 60 cases of patients 24 months found no HBsAg seroconversion, four cases of emergence of HBeAg seroconversion. CONCLUSION: (1) detection of serum HBV-LHBs to reflect the hepatitis B virus replication with HBVDNA good correlation. (2) anti-viral treatment of dynamic observation of the process of HBV-LHBs expression can predict the effectiveness of anti-viral therapy.

Induction of biliary cholangiocarcinoma cell apoptosis by 103Pd cholangial radioactive stent gamma-rays.

BACKGROUND: In recent years, interventional tumor therapy, involving implantation of intra-cholangial metal stents through percutaneous trans-hepatic punctures, has provided a new method for treating cholangiocarcinoma. (103)Pd cholangial radioactive stents can concentrate high radioactive dosages into the malignant tumors and kill tumor cells effectively, in order to prevent re-stenosis of the lumen caused by a relapsed tumor. The aim of the present study was to investigate the efficacy of gamma-rays released by the (103)Pd biliary duct radioactive stent in treating cholangiocarcinoma via induction of biliary cholangiocarcinoma cell apoptosis. METHODS: A group of biliary duct cancer cells was collectively treated with a dose of gamma-rays. Cells were then examined by the 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl terazolium-bromide (MTT) technique for determining the inhibition rate of the biliary duct cancer cells, as well as with other methods including electron microscopy, DNA agarose gel electrophoresis, and flow cytometry were applied for the evaluation of their morphological and biochemical characteristics. The growth curve and the growth inhibition rate of the cells were determined, and the changes in the ultrastructure of the cholangiocarcinoma cells and the DNA electrophoresis bands were examined under a UV-lamp. RESULTS: The gamma-ray released by (103)Pd inhibited cholangiocarcinoma cell growth, as demonstrated when the growth rate of the cells was stunned by a gamma-ray with a dosage larger than 197.321 MBq. Typical features of cholangiocarcinoma cell apoptosis were observed in the 197.321 MBq dosage group, while cell necrosis was observed when irradiated by a dosage above 245.865 MBq. DNA agarose gel electrophoresis results were different between the 197.321 MBq irradiation dosage group, the 245.865 MBq irradiation dosage group, and the control group. CONCLUSIONS: (103)Pd radioactive stents which provide a radioactive dosage of 197.321 MBq are effective in the treatment of cholangiocarcinoma; (103)Pd radioactive stents should be useful for the clinical treatment of cholangiocarcinoma.

[Determination of intestinal permeability in cholelithiasis patients by oral administration of technetium-99m-diethylenetriaminepentaacetatic acid].

OBJECTIVE: To investigate the intestinal permeability of patients with cholelithiasis of different types. METHODS: Technetium-99m-diethylenetriaminepentaacetatic acid (99mTc-DTPA) at the dose of 185 MBq (5 mCi) was administered orally to 56 patients of cholelithiasis, 15 cases of cholesterol stone (CS group) and 41 cases of pigment stone (PS group) based on the cross section of the stone during operation, and 17 healthy controls. A 24 h urine collection was obtained after the ingestion of the tracer to calculate the urinary excretion of DTPA. RESULTS: The mean percentage of the total ingested dose of 99mTc-DTPA excreted in a 24 h urinary excretion was 5.0%+/-3.6% in the CS group, not significantly different from that in the control group (4.5%+/-3.4%. F=2.18, P>0.05), and the mean percentage of the total ingested dose of 99mTc-DTPA excreted in a 24 h urinary excretion of the PS group was 10.5%+/-6.9%, significantly higher than that in the control group (F=7.62, P<0.05), showing a significantly increase of intestinal permeability (P<0.05). CONCLUSION: The intestinal permeability of the patients of pigment stone is higher than that of the healthy subjects. Hyperpermeability may be a factor of the pathogenesis of pigment stone.

[Transplantation of human limbal cells cultivated on amniotic membrane for reconstruction of rat corneal epithelium after alkali burn].

OBJECTIVE: To investigate the biological characters of limbal cells and evaluate the effect of cultivated human limbal epithelial cells transplantation on ocular surface reconstruction. METHODS: Human limbal cells were isolated and cultivated in vitro. Immunofluorescence staining and RT-PCR were used to study the phenotype of the cells, BrdU labeling test was used to identify the slow-cycling cells in the cultures. Limbal stem cell deficiency (LSCD) was established in rat cornea by alkali burn. Two weeks after the injury, the rats received transplantation of cultivated human limbal epithelial cells with amniotic membrane carrier, and then the therapeutic effects were evaluated by slit lamp observation, HE staining and immunofluorescent staining. RESULTS: On day 7, p63 and K19 were strongly expressed by most cells, only a few cells expressed K3. On day 14 and day 21, p63 and K19 were still expressed by a majority of cells, while the proportion of K3 positive cells increased, some cells co-expressed p63 and K3. RT-PCR showed that gene expression of both p63 and K12 were positive in cultivated limbal cells, but in mature superficial epithelial cells only K12 was detected. Slow-cycling cells were observed after cultured for 21 days with BrdU free medium. Four weeks after limbal stem cells combined amniotic membrane transplantation (LSAT), both slit lamp observation and HE staining showed that LSAT relieved the pathological changes of rat cornea notably as compared with the amniotic membrane transplantation (AMT) group and control group. The rats that received LSAT exhibited reconstructed corneas with the intact epithelium and improved transparency. Immunofluorescence staining showed that a majority of the rat corneal epithelial cells stained positively to anti-human nuclear antibody and K3 antibody. CONCLUSIONS: P63 is not exclusively expressed by limbal stem cells (LSCs), a certain amount of p63 may also expressed by transient amplifying cells, LSCs are identified as p63 and K19 positive, K3/K12 negative cells. The detection of slow-cycling cells in the culture confirms that LSCs can be cultivated in vitro. Cultivated LSCs combine with amniotic membrane transplantation can functionally reconstruct the cornea suffered with LSCD.

[Subject diagnostic value of detecting a1pha-fetoprotein variants with a new microspincolumn method in hepatocellular carcinoma].

OBJECTIVE: To evaluate the usefulness of new microspincolumn method for the measurement of a1pha-fetoprotein variant AFP-L3 in differentiation of benign and malignant liver disease and the warming for liver cancer. METHODS: AFP-L3 was isolated by using microspincolumn coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were determined with chemiluminescent immunoassay, the proportion of AFP-L3 levels AFP-L3(%) were calculated, and the relationship between the elevated AFP-L3(%) levels and benign and malignant liver disease was analyzed. RESULTS: The levels of AFP-L3(%) in serum of patients with hepatocellular carcinoma was significantly higher than those in the patients with other liver diseases (P < 0.001). Taking AFP-L3(%) >or= 10% as the diagnostic criteria, the sensitivity for diagnosis of liver cancer was 90.9%. CONCLUSION: Detection of AFP-L3 seemed to be of clinical value in diagnosis and differential diagnosis of hepatocellular carcinoma; it may be especially important for identifying patients with hepatocellular carcinoma whose a1pha-fetoprotein level is low.

Transplantation of human limbal cells cultivated on amniotic membrane for reconstruction of rat corneal epithelium after alkaline burn.

BACKGROUND: The transplantation of limbal epithelial cells cultivated on amniotic membrane is a newly developed treatment for limbal stem cell deficiency. The purpose of our study was to investigate the biological characteristics of limbal epithelial cells and evaluate the effect of transplantation of cultivated human limbal epithelial cells on ocular surface reconstruction in limbal stem cell deficiency rat model. METHODS: Human limbal cells were isolated and cultivated in vitro. Cytokeratins 3, 12, and 19 (K3, K12 and K19) and p63 were detected by immunofluorescent staining or RT-PCR. BrdU labelling test was used to identify the slow cycling cells in the cultures. Limbal stem cell deficiency was established in rat cornea by alkali burn. Two weeks after injury, the rats received transplants of human limbal stem cells cultivated on amniotic membrane carrier. The therapeutic effect was evaluated by slit lamp observation, Hemotoxin and Eosin (HE) staining and immunofluorescent staining. RESULTS: On day 7 in primary culture, p63 and K19 were strongly expressed by most cells but only a few cells expressed K3. On days 14 and 21, p63 and K19 were still expressed by a majority of cells, but the expressive intensity of p63 decreased in a number of cells, while the proportion of K3 positive cells increased slightly and some cells coexpressed p63 and K3. RT-PCR showed that gene expression of both p63 and K12 were positive in cultivated limbal cells, but in mature superficial epithelial cells, only K12 was detected. BrdU labelling test showed that most cells were labelled with BrdU after 7 days' labelling and BrdU label retaining cells were observed after chasing for 21 days with BrdU free medium. For in vivo test, slit lamp observation, HE staining and immunofluorescent staining showed that the rats receiving transplant of human limbal stem cells cultivated on amniotic membrane grew reconstructed corneas with intact epithelium, improved transparency and slight or no neovascularization. A majority of epithelial cells of the reconstructed cornea were positive to antihuman nuclear antibody and cells expressing K3 were found mainly in superfacial epithelium. CONCLUSIONS: Limbal stem cells can be cultivated in vitro: the cells are characterized by high proliferation and slow cycling and identified as p63/K19 positive and K3/K12 negative. During culture, some stem cells can proliferate and differentiate into mature cornea epithelial cells. Amniotic membrane is a suitable carrier for limbal stem cells. Transplantation of human limbal stem cells cultivated on amniotic membrane can functionally reconstruct rat cornea with limbal stem cell deficiency.

Diabetic retinopathy: VEGF, bFGF and retinal vascular pathology.

BACKGROUND: Previous research indicated that the development of diabetic retinopathy (DR) is closely related to the excessive expression of growth factors. This paper was to study the relationship of DR with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the retinal vascular pathological change. METHODS: Fifty-five Wistar rats, weighing 100 - 200 g, were selected and randomly divided into four groups: control group (no streptozocin injection, n = 10), M1 group (streptozocin induced diabetes for 1 month, n = 15), M3 group (streptozocin induced diabetes for 3 months, n = 15), and M5 group (streptozocin induced diabetes for 5 months, n = 15). In situ hybridization and immunohistochemistry were used to investigate the expressions of bFGF and VEGF on retinal vascular, and retinal vessels were observed by transmission electron microscope. RESULTS: There was no difference in the number of pericytes between M1 and control group (P > 0.05), but the number of pericytes decreased obviously in M3 and M5 groups compared with the control group (P < 0.01, P < 0.001, respectively). Capillary embolization and non-cell capillary were seen in M5 group. Positive expression of VEGF was found in M5 group using in situ hybridization and immunohistochemistry respectively. Positive expression of bFGF could be seen in M3 (78%) and M5 group (89%). Most remarkable changes of vessels were observed in M5 group including fragmental thickness, split of basement membrane, swelling and distortion of endothelial cells. CONCLUSIONS: In retinal vascular of the streptozocin (STZ) rats, there shows the expression of bFGF at the third month and that of VEGF at the fifth month.

[Effects of vascular endothelial growth factor and basic fibroblast growth factor in early stage diabetic retinopathy and their molecular pathological mechanism].

OBJECTIVE: To study the effects of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in early stage diabetic retinopathy and their mechanisms, so as to guide the clinical work theoretically. METHODS: Fifty-five Wistar rats were randomly divided into four groups. Ten rats were used as controls (M group). Streptozocin was injected intraperitoneally to induce diabetes for I month (M(1) group, n = 15), 3 months (M(3) group, n = 15), and 5 months (M(5) group, n = 15). At the experimental ends of each group, the rats were over-anesthetized and their eyeballs were extracted to make digest preparation. In situ hybridization and immunohistochemistry were used to investigate the expression of bFGF and VEGF on retinal vascular. Transmission electron microscopy was used to observe the histology of the retinal vessels. RESULTS: There was no difference in the number of pericytes between M(1) group and M group (P > 0.05), but the number of pericytes was significantly lower in M(3) and M(5) groups than in the M and M(1) groups (P < 0.001). VEGF in situ hybridization showed an expression rate of 34% in the M(5) group. VEGF immunohistochemistry showed an expression rate of 56% in M(5) group. bFGF in situ hybridization showed an expression rate of 78% in M(3) group and an expression rate of 89% in M(5) group. Transmission electron microscopy showed no change in M group. However, it showed swelling of endothelial cells, finger-like process into the capillary cavity, and uneven distribution of heterochromatin in pericytes in M(1) group. Obvious fragmental thickening and splitting of basement membrane, swelling and deformation, finger like process to the capillary cavity, and concentration and margination of heterochromatin in endothelial cells and swelling and deformation of rough surfaced endoplasmic reticulum and mitochondrion in pericytes were seen in the M(3) and M(5) groups, especially the latter. CONCLUSION: During the course of diabetic retinopathy, morphologic changes of vessels occurs prior to the expression of growth factors, in which the expression of VEGF follows the expression of bFGF.