To identify the pathogen of severe acute respiratory syndrome (SARS ), suckling mice and Vero E6 cells were inoculated with the lung samples from autopsied patients with SARS. The tissues obtained from mice were examined with light microscopy and electron microscope(TEM). The infected cells and supernatants were also examined under TEM. The primary hiatopathological leisions in the lungs were wall of lung alveolies thickened and alveolar cavities narrowed. The hepatocytes were swollen and undergone vacuolar and hydropic degeneration. A large quantity of coronavirus like particles could be observed in the infected cells and supernatants and also in mouse pneumocytes under TEM. The viruses are round with diameters 80～120nm.The isolated coronavirus was associated with the current SARS epidemics.

OBJECTIVE To investigate the resistance of Acinetobacter baumannii(ABA),and the mechanism of imipenem resistance in A.baumannii.METHODS All specimens were identified by VITEK-60 and the drug resistance was detected by Kirby-Bauer test.Three dimensional test was used to detect ESBLs and AmpC.PCR and DNA sequencing were performed to determine VIM,IMP,OXA-23 and OXA-24 ?-lactamases.Outer membrane protein was analyzed by SDS-PAGE,Reserpine synergistic inhibition test was used to study the active efflux mechanism.RESULTS Totally 120 strains were isolated from sputum(76.9%),and 16 strains from secretion(10.3%).ICU was the main infected department(51 strains,32.7%).The resistance to sulperazone was the lowest(20.5%),and to imipenem accounted for 38.5%,Of the 20 imipenem resistant strains,10 strains were ESBLs positive(50%),and 20 strains were AmpC positive(100%).VIM,IMP and OXA-24 ?-lactamases were not detected out,19 strains(95%) produced OXA-23.Compared to the imipenem-sensitive strains,the resistant strains lacked the outer membrane protein of 22?103,29?103 and 33?103.The MICs of A.baumannii to imipenem were not decreased by reserpine which demonstrated that excretive mechanism was negative.CONCLUSIONS ICU is the main infected department for ABA.The resistance rate is increasing for longer-term usage of carbopenem;OXA-23 production is the important resistance mechanism in ABA,AmpC production and outer membrane protein lacking show close relation to the drug-resistance in A.baumannii.

Objective To explore the effect of cryopreserved canine kidney cells (MDCK)single -layer on the isolation and proliferation of influenza virus B.Methods Revived P17 MDCK cells were passage for 2 -4 gener-ations,and subsequently preserved in 4℃ refrigerator for 3,6 and 9 days,respectively.Under same experimental con-ditions,the 4℃ refigerater preserved cells were co -incubated with influenza -like illness(ILI)throat swab speci-mens.Cytopathic effect (CPE)was observed,and the proliferation of virus was determined using real -time PCR and the hemagglutinin titers were determined by serological test.Results (1)CPE:The CPE of the MDCK cells pre-served in 4℃ refrigerator for 3 or 6 days had no significant differences compared with that in the control group,while the cell preserved in 4℃ refrigerator for 9 days showed CPE fastly and maintained for a short time.(2)Real -time PCR:the proliferation of influenza virus B in the MDCK cells preserved with 4℃refrigerator for 3 or 6 days (25.86 × 105 -30.25 ×106 ,26.31 ×105 -30.54 ×106 )had on difference compared with that of the control group (24.82 × 105 -29.86 ×106 ),with the proliferation rate of 105 to 106 times,while the proliferation cell with 4℃ cryopreserved for 9 days(19.72 ×104 -28.34 ×105 )the proliferation in cells preserved for 9 days was sharply decreased,with pro-liferation rate of 104 to 105 times.(3)The HA titer:The virus strains with hemagglutination titer above or equal to 116 (P >0.05)isolated with MDCK cells preserved in 4℃ refrigerator 3 or 6 days were not significantly different from that of the control group (10.92 ±0.79).And the cells with 4℃ cryopreserved for 9 days were significantly dicreased (P <0.01).Conclusion No significant effects on the isolation and proliferation of influenza virus B using MDCK cell preserved in 4℃ frigerator for near one week were observed in the present study.

Objective To investigate the role of hemoglobin (Hb),red blood cell count (RBC) and hematocrit (HCT) during the first trimester in predicting the occurrence of gestational diabetes mellitus (GDM) in the third trimester in nonanemic women in Kunming.Methods A prospective study was carried out.Routine blood tests were performed in 1189 nondiabetic singleton pregnant Chinese women at their initial prenatal healthcare before 14 gestational weeks in the First Affiliated Hospital of Kunming Medical University from January 1,2008 to December 31,2009.They were divided into four groups by quartiles of Hb,RBC and HCT levels respectively.Differences of GDM incidence at the third trimester were compared among groups.Kollmogorov Smirnov test was used to detect normal distribution of Hb,RBC and HCT results.Differences between rates of groups were detected by Chi-square test.Independent risk factors of GDM were screened by Logistic regression analysis.Results The incidence of GDM at the third trimester increased as quartile categories of Hb,RBC and HCT ascended.The GDM incidence (13.1 ％,34/259) of ≥75th Hb group was higher than that of ＜ 25th group (5.9 ％,18/304,x2 =8.53,P＜ 0.0 1),25th ～ group (9.2 ％,30/326,x2=4.18,P＜0.05) and 50th～ group (9.3％,28/300,x2=3.96,P＜0.05).The GDM incidence (11.7 ％,34/290) of 50th～ RBC group (RBC≥4.44 × 1012/L) was higher than that of ＜25th group (6.3％,19/304,x2=5.30,P＜0.05),25th～ group (7.0％,21/298,x2 =3.93,P＜0.05); and the GDM incidence (12.1％,36/297) of ≥75th group was also higher than that of ＜25th group (6.3％,19/304,x2 =6.49,P＜0.05),25th～ group (7.0％,21/298,x2=3.85,P＜0.05).The GDM incidence of ＜25th HCT group (5.3％,16/303) was lower than that of 25th～ group (9.3％,29/311,x2=7.04,P＜0.01),50th～ group (10.8％,31/287,x2=3.93,P＜0.05) and ≥75th group (11.8％,34/288,x2=6.49,P＜0.05).Logistic regression analysis showed that Hb (OR=1.031,95％ CI:1.010-1.052),RBC (OR=2.286,95％CI:1.318-3.963) and HCT (OR=1.106,95％ CI:1.037-1.179) were independent risk factors of GDM (all P＜0.05).Conclusions High maternal Hb,RBC and HCT levels in the first trimester could be used in predicting GDM in pregnant women in Kunming area.

Objective To evaluate the best mean for the treatment of varicocele. Methods The clini-cal effects of the high ligation of spermatic vein under the small incisal opening of the poritneum meta abdom-inal membrane and the ligation of spermatic vein under the laparoscope for the varicocele were retrospectively reviewed. The operating time, the postoperative days and the cost of hospitalization were compared of the two methods. Results Comparing with the group of operation and the group of laparoscope. Among the 158 vat-icocele patients with unilateral or bilateral varicocele, there was not statistical difference in the operating time between the two methods (P > 0. 05), whi]e there was significant difference in the postoperative days and the cost of hospitalization(P <0. 05). Conclusions When the bilateral varicoceles were simultaneously ligated, the high ligation of spermatic vein under the laparoscope was the best selection; if the unilateral varicocele was highly ligated or the patients were unconditional or the patients were not fit for laparoscopic operation, the high ligation of spermatic vein under the small incisal opening of peritoneum was the best operating method.

The protein production system using a baculovirus Antheraea pernyi nucleopolyhedrovirus (AnpeNPV) as a gene expression vector and its host insect as a natural bioreactor was successful established and its excellent performance in the protein production has been demonstrated. In this paper, the system is used to produce recombinant human epidermal growth factor (rhEGF), which have been widely used in medical and cosmetic treatment. A recombinant AnpehEGF virus has been constructed by replacing the viral polyhedrin gene with the rhEGF gene, and then injected it to Samia cynthia ricini pupae. Amplification and expression of rhEGF gene in the pupae was clearly detected by PCR, Western blot and ELISA analyses. These analyses have also revealed that rhEGF in the pupae was significantly increased at 6 days post-infection, and reached maximum level at the 12th day. The concentrations of rhEGF were 19.77, 24.90, 618.59 and 1 952.46 ng/g pupae at 3, 6, 9 and 12 days post-infection, respectively. However, the rhEGF concentration reduced at later stage (days 15). The rhEGF in the pupae could be purified using ammonium sulfate precipitation and Ni-NTA agrose affinity chromatography. Results demonstrate that Samia cynthia ricini pupae can be used as a bioreactor to produce rhEGF and, if successfully improved, will be a novel method of rhEGF production with lower cost and more efficient.
Amino Acid Sequence ; Animals ; Bombyx ; genetics ; metabolism ; Epidermal Growth Factor ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Humans ; Molecular Sequence Data ; Nucleopolyhedrovirus ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

Objective To investigate the molecular characteristics of H5 subtype avian influenza viruses (AIV) in Weining, Guizhou Province. Methods Nine representative strains were randomly select-ed from H5 subtype AIV that were identified by real-time PCR in Weining, Guizhou Province from 2015 to 2017. Nucleic acid was extracted from each sample and hemagglutinin (HA) genes were amplified and then sequenced. Homology, genetic evolution and the sites related to pathogenicity, receptor binding regions as well as potential glycosylation of H5 AIV were analyzed by bioinformation software. Results Homology analysis revealed that there was 96. 1%-99. 9% and 95. 7%-100% similarity among the nine strains in nu-cleotide and amino acid of HA gene, respectively. These strains belonged to two branches, H5-1 and H5-2. The cleavage site motifs were PLREKRRKR↓GLF for five strains in H5-1 branch and PQRERRRKR↓GLF for four strains in H5-2 branch, which made them high pathogenic. QSG and QRG at the key receptor bind-ing sites were found in H5-1 and H5-2 branch strains, respectively. They were responsible for receptor bind-ing specificity of AIV. Mutations of 138Q, 139G and 53K were all detected in the nine strains. 129K, 189T, 140K and 282V mutations were discovered in the five strains of H5-1 branch, while 189N, 140M and 282I mutations were found in the four strains of H5-2 branch. Results of the glycosylation motif analysis showed that six sites were conservative, but there was an addition of 124NHT site in two strains of H5-2 branch isolated in 2017. Conclusion Two high pathogenic H5 subtypes of AIV could be epidemic in Wein-ing, Guizhou Province during 2015 to 2017. Although H5 subtype AIV did not possess specific receptor binding regions like human influenza viruses, they were in continuous variation with an increase in glycosyla-tion motifs, which might enhance their virulence and pathogenicity to human beings. Hence, surveillance and study on the molecular properties of H5 subtype AIV should be strengthened.

【Objective】 To report the early clinical effects and surgical complications of endoscopic spinal fusion technology （Endo-LIF） in the treatment of degenerative lumbar disease. 【Methods】 The clinical data of 31 patients with degenerative lumbar spine disease treated with Endo-LIF from June 2019 to May 2021 were retrospectively analyzed. All the 31 patients underwent endoscopic spinal fusion therapy. We recorded the operation time， hospital stay duration， postoperative complications， visual analogue scale for pain （VAS）， oswestry dysfunction Index （ODI） and low back pain in the Chinese Orthopaedic Association Spine Group Surgery scoring standards before operation， immediately after operation， and the last follow-up to evaluate clinical efficacy. 【Results】 The operation time of the 31 patients was （134.80±34.98） min， the intraoperative blood loss was （100.13±18.49） mL， the hospital stay was （6.65±0.17） days， and the follow-up time was 6 to 18 （14±2.3） months. One patient had hematoma compression after surgery； he had incision made immediately to clear the hematoma and healed after bed rest. Two patients developed spinal hypertension and healed after bed rest. All the patients had no symptoms of nerve injury after operation， and the clinical symptoms were significantly relieved. We compared the perioperative VAS score and ODI index of all the patients， which were lower immediately after operation and at the last follow-up than those before the operation （P<0.05）， and the difference was statistically significant. 【Conclusion】 Endo-LIF technology has good short-term clinical effects and the advantages of milder trauma， less blood loss， and quick recovery after surgery. It is a safe and minimally invasive lumbar fusion surgery.