Clinical Characteristics of Recurrent-positive Coronavirus Disease 2019 after Curative Discharge: a retrospective analysis of 15 cases in Wuhan China

In China, the patients with previously negative RT-PCR results again test positive during the post-discharge isolation period. We aimed to determine the clinical characteristics of these "recurrent-positive" patients. We retrospectively reviewed the data of 15 recurrent-positive patients and 107 control patients with non-recurrent, moderate COVID-19 treated in Wuhan, China. Clinical data and laboratory results were comparatively analyzed. We found that recurrent-positive patients had moderate disease. The rate of recurrent-positive disease in our hospital was 1.87%. Recurrent-positive patients were significantly younger (43(35-54) years) than control patients (60(43-69) years) (P=0.011). The early LOS (length of stay in hospital before recurrence) was significantly longer in recurrent-positive patients (36(34-45) days) than in control patients (15(7-30) days) (P =0.001). The time required for the first conversion of RT-PCR results from positive to negative was significantly longer in recurrent-positive patients (14(10-17) days) than in control patients (6(3-9) days) (P =0.011). Serum COVID-19 antibody levels were significantly lower in recurrent-positive patients than in control patients (IgM: 13.69 {+/-} 4.38 vs. 68.10 {+/-} 20.85 AU/mL, P = 0.015; IgG: 78.53 {+/-} 9.30 vs. 147.85 {+/-} 13.33 AU/mL, P < 0.0001). Recurrent-positive patients were younger than control patients. The early LOS (length of stay in hospital before recurrence) was significantly longer in recurrent-positive group than that in control group. COVID-19 IgM/IgG antibody levels were significantly lower in recurrent-positive group than those in control group, which might explain why the virus RNA RT-PCR was positive after the initial "clinical cure"(with three times of virus RNA RT-PCR negative). The virus might not be fully eliminated because of the lower IgG level and their later replicating might result in recurrent-positive virus RNA RT-PCR.

Efficacy and safety of chloroquine or hydroxychloroquine in moderate type of COVID-19: a prospective open-label randomized controlled study

The outbreak of novel coronavirus disease 2019 (COVID-19) has become a pandemic. Drug repurposing may represent a rapid way to fill the urgent need for effective treatment. We evaluated the clinical utility of chloroquine and hydroxychloroquine in treating COVID-19. Forty-eight patients with moderate COVID-19 were randomized to oral treatment with chloroquine (1000 mg QD on Day 1, then 500 mg QD for 9 days; n=18), hydroxychloroquine (200 mg BID for 10 days; n=18), or control treatment (n=12). Adverse events were mild, except for one case of Grade 2 ALT elevation. Adverse events were more commonly observed in the chloroquine group (44.44%) and the hydroxychloroquine group (50.00%) than in the control group (16.67%). The chloroquine group achieved shorter time to clinical recovery (TTCR) than the control group (P=0.019). There was a trend toward reduced TTCR in the hydroxychloroquine group (P=0.049). The time to reach viral RNA negativity was significantly faster in the chloroquine group and the hydroxychloroquine group than in the control group (P=0.006 and P=0.010, respectively). The median numbers of days to reach RNA negativity in the chloroquine, hydroxychloroquine, and control groups was 2.5 (IQR: 2.0-3.8) days, 2.0 (IQR: 2.0-3.5) days, and 7.0 (IQR: 3.0-10.0) days, respectively. The chloroquine and hydroxychloroquine groups also showed trends toward improvement in the duration of hospitalization and findings on lung computerized tomography (CT). This study provides evidence that (hydroxy)chloroquine may be used effectively in treating moderate COVID-19 and supports larger trials.

Influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine / 药学学报

To investigate the influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine (DEN), a total of 40 rats were randomly divided into 4 groups: normal control group, model group, and two bear bile treatment groups. The rat liver cancer model was induced by breeding with water containing 100 mg x L(-1) DEN for 14 weeks. The rats of the bear bile groups received bear bile powder (200 or 400 mg x kg(-1)) orally 5 times per week for 18 weeks. The general condition and the body weight of rats were examined every day. After 18 weeks the activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and total bilirubin (TBIL) were detected. Meanwhile, the pathological changes of liver tissues were observed after H&E staining. The expression of proliferative cell nuclear antigen (PCNA) and a-smooth muscle actin (alpha-SMA) in liver tissue were detected by immunohistochemical method. After 4 weeks the body weights of rats in normal group were significantly more than that in other groups (P < 0.05); and that in the two bile groups was significantly more than that in the model group. Compared with normal group, the level of serum glutamic-pyruvic transaminase and total bilirubin increased significantly in other groups; compared with model group, these two indexes decreased significantly in two bile groups. Hepatocellular carcinoma occurred in all rats except for normal group; there were classic cirrhosis and cancer in model group while there were mild cirrhosis and high differentiation in two bile groups. There were almost no expressions of PCNA and alpha-SMA in normal group while there were high expressions in model group; the two bile groups had some expressions but were inferior to the model group, and alpha-SMA reduced markedly. It indicated that bear bile restrained the development of liver cancer during DEN inducing rat hepatocarcinoma, which may be related to its depressing hepatic stellate cell activation and relieving hepatic lesion and cirrhosis.

Immunocharacteristics of bone marrow mesenchymal stem cell / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the immunocharacteristics of bone marrow mesenchymal stem cell (MSC) and provide experimental evidence for the potential therapeutic application.</p><p><b>METHODS</b>MSCs were isolated from rat bone marrow and confirmed by immunophenotype, and the growth dynamic and cell cycle were analyzed. MSCs were cultured with or without 200 U/ml interferon gamma (IFNg) , the expression of PDL-1, CD54, CD40, CD80, CD86, MHC-I, and MHC-II was detected by flow cytometry. MSCs were used as regulatory cells in mixed lymphocyte reaction (MLR), the PDL-1 and CD54 molecules on MSCs were blocked to explore their roles in MLR. The IFN, IL-2, IL-4 and IL-10 molecules in culture supernatant were quantified by ELISA. The homing of MSCs to liver and induction of microchimerism were analyzed after MSCs transplantation.</p><p><b>RESULTS</b>The purity of MSCs was high. The growth curve showed that the first two days were the lag phase; the third, fourth, fifth days were the log phase; the sixth and seventh days were the stationary phase. Flow cytometry indicated that 76.0%+/-2.0% of the MSCs were in G1/G0 phase, 13.0%+/-2.0% in S phase, 10.0%+/-1.7% in G2 and M phase. IFNg treatment led to up-regulation of CD54, PDL-1, MHC-I and MHC-II, however, CD40, CD80 and CD86 were not expressed on MSCs even after IFNg treatment. MSCs inhibited MLR, IFNg treatment enhanced the inhibitory effect of MSCs on MLR. Blocking of PDL-1 or CD54 on MSCs partially alleviated the inhibition effect. There were high levels of IFNg and IL-10, and low level of IL-4 in the culture supernatant of MLR, however, IL-2 was not detected. MSCs can home to the liver and induce formation of microchimerism after transplantation.</p><p><b>CONCLUSION</b>IFNg treatment enhances the inhibitory effect of MSCs on MLR, PDL-1 and CD54 are key molecules mediating this inhibitory effect. MSC can home to the liver and induce formation of microchimerism after transplantation.</p>

Pathological monitoring of intestinal graft acute rejection in auxiliary en-bloc liver-small bowel transplantation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the pathologic monitoring of intestinal graft rejection in auxiliary en-bloc liver-small bowel transplantation in pigs.</p><p><b>METHODS</b>Fifty outbred long-white pigs were randomized into three groups, and the auxiliary composite liver-small bowel allotransplantations were undertaken in 10 pigs in group A and group B while segment small bowel allotransplantations were undertaken in 10 pigs in group C. Group A and C were not treated with immunosuppressive drugs while group B was treated with cyclosporine A and methylprednisolone. The postoperative intestinal graft rejections were monitored by biopsy through the jejunostomy or ileuostomy on 1, 3, 5, 7, 14, 21 and 30 days after operation. Through routine management, the specimens were directly examined via optical and electronic microscope respectively.</p><p><b>RESULTS</b>As shown from pathological data, the median initial time of postoperative rejection in group A was 8 days (ranged from 7 to 12), later than that in group C (5 days:ranged from 3 to 5), P<0.05). On the 7th day postoperatively, the rejection scores in group A was 1.11+/-0.20, lower than that in group C(2.56+/-0.18, P<0.05), but higher than that in group B(0.20+/-0.13, P<0.05). Ultrastructure also showed more severe intestinal graft rejection in intestinal transplantation than that in combined transplantation. The median survival time was 9 days(ranged from 7 to 25) in group A and 12 days(ranged from 7 to 20) in group C, while all the pigs in group B lived longer than 30 days.</p><p><b>CONCLUSION</b>The pathological assessment through the jejunostomy or ileuostomy biopsy is a convenient method to monitor the postoperative graft rejections in intestinal related transplantation.</p>