Objective To investigate the diagnostic value of microscopic observation drug susceptibility assay (MODS) in extrapulmonary tuberculosis. Methods MODS technology was constructed by using 24-well cell culture plate and liquid culture.Ziehl-Neelsen smear,Lowenstein-Jensen culture and MODS were used to detect Mycobacterium tuberculosis in 74 pleural fluid samples collected from patients with tuberculous pleurisy,63 cerebrospinal fluid samples collected from patients with tuberculous meningitis and 18 samples collected from non-tuberculosis suspects.The immunochromatography was used to distinguish Mycobacterium tuberculosis from nontuberculosis mycobacteria. The results of Ziehl-Neelsen smear, Lowenstein-Jensen culture and MODS were compared by x2 test.Results The positive rates of MODS,Lowenstein-Jensen culture and Ziehl-Neelsen smear were 58.1 ％ (43/74),18.9 ％ (14/74 ) and 6.8％ (5/74),respectively in tuberculous pleurisy patients; 54.0％(34/63),20.6％ (13/63) and 4.8％ (3/63),respectively in tuberculous meningitis patients.The positive rate of MODS technology was significantly higher than that of Lowenstein Jensen culture in tuberculous pleurisy patients (x2 =24.00,P＜0.01) and tuberculous meningitis patients (x2 =14.97,P ＜ 0.01). Each Mycobacterium obtained from MODS and Lowenstein-Jensen culture was identified as Mycobacterium tuberculosis by immunochromatography.All of the 18 pleural fluid and cerebrospinal fluid samples which collected from non-tuberculosis suspects were all negative detected by Ziehl-Neelsen smear,Lowenstein-Jensen culture and MODS.The median time to culture positive of MODS was 9 days in cerebrospinal fluid and 14 days in pleural fluid samples,which were both significantly shorter than that of Lowenstein-Jensen culture (31 days in both cerebrospinal fluid and pleural fluid samples). Conclusion Compared to conventional microbiological diagnosis methods,MODS is a rapid detection method of Mycobacterium tuberculosis with a higher positive detection rate,which is suitable for rapid diagnosis of extrapulmonary tuberculosis.

Objective To explore the mechanism and regulative function of betulin on anti-aging genes related of normol human dermal fibroblasts (ESF-1), and to reveal the repairing mechanism of betulin on wrinkles. Methods The cultured cell in vitro were divided into the control group, the estradiol group and the betulin high, medium and low dose groups. Competence of ESF-1 cells was detected by MTT. Expression of mRNA of collagen type I and Ⅲ (ColⅠand Col Ⅲ), tissue inhibitors of matrix metalloproteinase 1 and 2 (TIMP-1 and TIMP-2), matrix metalloproteinase 1 (MMP-1) was detected by RT-PCR. Results Compared with the control group, the proliferation of ESF-1 cells (0.455 ± 0.037, 0.445 ± 0.040 vs. 0.385 ± 0.601) in the estradiol group and the medial-dose betulin group were increased (P<0.05). Compared with the control group, the expression of mRNA of ColⅠ (0.960 ± 0.012, 0.929 ± 0.015 vs. 0.842 ± 0.014), Col Ⅲ (0.892 ± 0.009, 0.824 ± 0.022 vs. 0.768 ± 0.025), TIMP-1 (0.938 ± 0.026, 0.878 ± 0.035 vs. 0.796 ± 0.022), TIMP-2 (0.557 ± 0.025, 0.506 ± 0.036 vs. 0.436 ± 0.063) in the estradiol group and the medial-dose betulin group were increased (all P<0.01). Conclusion Betulin based on increasing the competence of ESF-1 cells, promoting collagen synthesis and the expression level of related-proteinase is to suspend the development of wrinkles.

Objective:To evaluate the efficacy and safety of cytoreductive surgery (CRS) in conjunction with hyperthermic intra-peritoneal chemotherapy (HIPEC) for treating patients with peritoneal carcinomatosis (PC) from colorectal cancer (CRC). Methods:A total of 62 CRC patients with complication of PC were divided into the CRS group, namely, Group One (n=29, CRS and systemic adju-vant chemotherapy) and the CRS+HIPEC group, namely, Group Two (n=33, CRS+HIPEC). The primary end point of the study was overall survival (OS) and the secondary end point was serious adverse events (SAE). Results:Patients' clinicopathologic characteris-tics, peritoneal carcinomatosis index, and completeness of cytoreduction therapy were well balanced and comparable between the two groups. The median follow-up was 41.9 mo (6.5 mo to 110.0 mo) in Group One and 32.0 mo (10.5 mo to 95.9 mo) in Group Two. The median OS was 8.5 mo (95%CI:4.9 mo to 12.1 mo) in Group One and 14.5 mo (95%CI:11.9 mo to 17.1 mo) in Group Two (P=0.007). Within 30 days after the surgery, SAE occurred in 3 of the 29 patients in Group One, and 9 of the 33 patients in Group Two (P=0.126). Multivariate analysis revealed that HIPEC, CC0-1 score, and chemotherapy over six cycles were the independent factors for OS improvement. Conclusion:The CRS+HIPEC method improves the OS of patients with PC from CRC, suggesting an acceptable safety.

This is a report of 3 cases of parkinsonism with long term administration of lamivudine. The clinic features were mask like faces, shuffling gait, lethargy and reduced automatic movement. After stopping use of lamivudine and treatment of anticholingeric drugs, the symptoms and signs of all cases were ameliorated. The possibilities of Wilson’s disease and other secondary parkinsonisms were excluded. The parkinsonism was attributed to complications from lamivudine. Lamivudine associated parkinsonism has not been reported previously in the medical literature.

Acute gastrointestinal dysfunction is a common and important complication of sepsis. As no exiting formal definition and classification of gastrointestinal dysfunction, most of the treatment strategies for gastrointestinal dysfunction are not based on clinical evidence, but on their own clinical experience. Experts of traditional Chinese medicine, integrated traditional Chinese and Western medicine and Western medicine from various disciplines in Shanghai are organized by the Shanghai Society of Integrated Traditional Chinese and Western Medicine and the Emergency Department Branch of Shanghai Physicians Association. After repeated discussion, literature search and formulation of the outline, we developed consensus on gastrointestinal dysfunction secondary to sepsis with integrating Traditional Chinese Medicine and Western medicine by consulting extensively on clinical experts in the fields of emergency medicine, gastroenterology, general surgery, infectious medicine and traditional Chinese medicine, and holding several expert forums and consultation meetings. This clinical expert consensus focused on acute gastrointestinal injury (AGI) classification and inducer of sepsis. In this consensus, the common symptoms, diagnosis, classifications, treatment strategies and suggestions of acute gastrointestinal injury or dysfunction secondary to sepsis were explored from the aspect of both Traditional Chinese Medicine and Western medicine.

Objective To investigate the serological and molecular identification of 2 rare B( A) blood groups. Methods The ABO blood groups of 2 samples from blood donors were detected by routine serological method. The genotype features was identified by PCR-sequence specific primer (PCR-SSP) and direct sequence analysis. Results The serological results for the 2 blood donors showed the characteristics of B(A) phenotype. The sample 1 was genotyped as BO2 subtype by PCR-SSP and direct sequencing showed B alleles in exon 7, presented nt640 A＞G mutation which was confirmed to be B(A)04/O02 genotype.The sample 2 was genotyped as BO1 sub-type by PCR-SSP and direct sequencing showed B alleles presented nt700 C＞G mutation in exon 7 which was confirmed to be B(A)02/O01 genotype. Conclusion The phenotype of the two samples should be B ( A ) and the genotypes should be rare B(A)04/O02 and B(A)02/O01.

Objective:To compare the clinical efficacies between arthroscopic anatomic single bundle reconstruction of posterior cruciate ligament (PCL) assisted by internal tension relieving technique combined with rapid rehabilitation and anatomic single bundle reconstruction combined with conventional rehabilitation in the treatment of PCL rupture.Methods:A retrospective cohort study was used to analyze the clinical data of 88 patients with PCL rupture admitted to First Affiliated Hospital of Kunming Medical University from September 2016 to September 2020. The patients included 65 males and 23 females, aged 18-55 years [(39.3±10.8)years]. Forty-four patients underwent arthroscopic anatomic single bundle reconstruction of PCL assisted by internal tension relieving technique combined with rapid rehabilitation therapy (tension-relieving group), and 44 patients underwent arthroscopic routine anatomic single bundle reconstruction of PCL combined with conventional rehabilitation (traditional group). The two groups were compared before and at 3, 12, and 24 months after surgery regarding the following items: International Knee Literature Committee (IKDC) score, Hospital for Special Surgery (HSS) score, Lysholm score, knee motion cycle (maximum stride length, minimum stride length, and stride frequency) and 6 kinematic indicators (angle of forward and back extension, angle of internal and external rotation, angle of internal and external rotation, up and down displacement, internal and external displacement, and forward and back displacement). The Marburger arthroscopy score (MAS) and Professor Ao Yingfang′s improved score by secondary arthroscopy were compared between the two groups at 12 months after surgery. The perioperative complications were observed.Results:All patients were followed for 24-36 months [(25.5±6.3)months]. In tension-relieving group and the traditional group, the values of IKDC score were (71.8±9.8)points and (68.5±6.5)points at 3 months after surgery, (87.6±6.0)points and (87.6±5.5)points at 12 months after surgery, and (95.5±3.1)points and (92.8±11.6)points at 24 months after surgery, respectively. The values were gradually increased, significantly higher than those before surgery [(48.1±16.9)points and (47.1±15.0)points] (all P<0.05). There were no significant differences between the two groups at each time point (all P>0.05). In tension-relieving group and the traditional group, the values of HSS score were (74.2±6.2)points and (68.4±9.5)points at 3 months after surgery, (91.9±5.4)points and (88.4±4.7)points at 12 months after surgery, and (97.1±2.0)points and (96.2±2.8)points at 24 months after surgery, respectively. The values of HSS score gradually increased, significantly higher than those before surgery [(57.5±17.7)points and (56.8±14.3)points] (all P<0.05). At 3 and 12 months after surgery, the values of HSS score in the tension-relieving group were significantly higher than those in the traditional group (all P<0.05), but others were not significantly different between the two groups (all P>0.05). In tension-relieving group and the traditional group, the values of Lysholm score were (74.2±14.9)points and (70.3±7.5)points at 3 months after surgery, (90.9±6.1)points and (88.7±4.7)points at 12 months after surgery, and (96.9±3.0)points and (96.3±2.8)points at 24 months after surgery, respectively. The values of Lysholm score were gradually increased, significantly higher than those before operation [(48.7±20.7)points and (48.2±19.9)points] (all P<0.05). There were no significant differences between the two groups at any time points (all P>0.05). At 3, 12, and 24 months after surgery, the motion cycle (maximum stride length, minimum stride length and stride frequency) and 6 kinematic indicators (angle of forward bending and backward extension, angle of internal and external rotation, angle of internal and external rotation, internal and external displacement, up and down displacement, and forward and backward displacement) of knee joint were significantly improved in both groups compared with those before surgery (all P<0.05). At 3, 12, and 24 months after surgery, the forward and backward displacement in the tension-relieving group was significantly decreased than that in the traditional group (all P<0.05), but others were not significantly different between the two groups (all P>0.05). The MAS was rated as excellent to good in 14 patients and fair to poor in 2, with the excellent and good rate of 87.5% (14/16) in the tension-relieving group, while the score was rated as excellent to good in 11 patients and fair to poor in 3, with the excellent and good rate of 78.6% (11/14) in the traditional group ( P>0.05). The Professor Ao Yingfang′s improved score was (10.6±1.5)points in the tension-relieving group, markedly higher than that in the traditional group [(9.6±2.3)points] ( P<0.05). No perioperative complications were observed. Conclusion:Compared with anatomic single bundle reconstruction combined with conventional rehabilitation, arthroscopic anatomic single bundle reconstruction of PCL assisted by internal tension relieving technique combined with rapid rehabilitation have higher subjective function score, better anteroposteric stability during knee movement, and better results of secondary microscopy.

BACKGROUND:Currently,there have been studies on the regulatory mechanism of lncRNA\miRNA\mRNA co-expression network on the occurrence and development of osteoarthritis.Our research group has screened qualified NONHSAT248596.1 and miR-146a-5p through the database in the previous stage,but the corresponding in vivo experiments to verify the above regulatory mechanisms are still lacking. OBJECTIVE:To explore the role of NONHSAT248596.1 in regulating competitive endogenous RNA of miR-146a-5p in cartilage degeneration mediated by stromal cell derived factor type 1/chemokine receptor 4 axis in vivo. METHODS:The models of osteoarthritis were established in 36 New Zealand rabbits by injecting stromal cell derived factor 1 solution into the knee joint of the right hind limb.According to the random number table method,they were divided into four groups.lncRNA group,miRNA group,ceRNA group and control group were injected with lentivirus vector overexpressing NONHSAT248596.1,lentivirus vector overexpressing miR-146a-5p,lentivirus vector overexpressing miR-146a-5p+NONHSAT248596.1 and empty lentivirus vector into the molded knee joint,respectively.At 4,8 and 12 weeks of modeling,cartilage tissues and subchondral bone tissues of the knee joint were taken for relevant detection. RESULTS AND CONCLUSION:Hematoxylin-eosin staining and safranin fast green staining showed different degrees of degeneration in the four groups.At 4 weeks,the cartilage tissue of the lncRNA group showed swelling of chondrocytes,loss of cell polarity,destruction of extracellular matrix,surface erosion,fracture formation and partial or full layer loss of cartilage tissue.The degree of cartilage injury was gradually aggravated with time.The progression of articular cartilage inflammation in the miRNA group was the slowest among the four groups.qRT-PCR showed that at the same time point,mRNA expression levels of NONHSAT248596.1,chemokine receptor 4,matrix metalloproteinase 3,matrix metalloproteinase 9 and matrix metalloproteinase 13 in cartilage tissue of the lncRNA group were higher than those of the other three groups(P＜0.05).The mRNA expression levels of miR-146a-5p,aggrecan and type Ⅱ collagen were lower than those of the other three groups(P＜0.05).The mRNA expression levels of NONHSAT248596.1,chemokine receptor 4,matrix metalloproteinase 3,matrix metalloproteinase 9 and matrix metalloproteinase 13 in the miRNA group were lower than those in the ceRNA group and control group at 8 and 12 weeks after the model construction(P＜0.05).The mRNA expressions of miR-146a-5p,aggrecan and type Ⅱ collagen were higher than those of the ceRNA group and control group(P＜0.05).Western blot assay showed that at the same time point,the expression levels of aggrecan and type Ⅱ collagen in cartilage tissue of the lncRNA group were always lower than those of the other three groups(P＜0.05).The expression levels of aggrecan and type Ⅱ collagen in cartilage tissue of the miRNA group at 8 and 12 weeks after modeling were higher than those of the ceRNA group and control group(P＜0.05).The results showed that miR-146a-5p,as the target of NONHSAT248596.1,could be inhibited by the effect of its ceRNA.After acting on miR-146a-5p,NONHSAT248596.1 regulates the stromal cell derived factor type 1/chemokine receptor 4 axis to affect the expression of matrix metalloprotein,type Ⅱ collagen,and aggrecan in osteoarthritis chondrocytes,resulting in the degradation of extracellular matrix and the loss of proteoglycan.

Cadmium (Cd) is a common heavy metal in the environment. Cd²⁺ may penetrate the blood-brain barrier and produce neurotoxicity, thus inducing various neurodegenerative diseases. Celastrol is an effective component of Tripterygium wilfordii Hook. F., which has many pharmacological effects such as anti-cancer and anti-inflammatory. Here we explored the effect of celastrol on the corresponding neurotoxicity induced by Cd²⁺. Cell proliferation test, cell membrane integrity test, and cell morphology were observed to analyze the effect of Cd²⁺ on the viability of HMC3. The neurotoxicity of Cd²⁺ and the effect of celastrol on the corresponding neurotoxicity induced by Cd²⁺ were analyzed by nitric oxide (NO) test, lipid peroxidation (MDA) test, and Western blotting. When the concentration of Cd²⁺ reached 40 μmol/L, the inhibition rate of HMC3 cell proliferation was (57.17±8.23)% (P < 0.01, n=5), compared with the control group. The cell activity continued to reduce when the Cd²⁺ concentration further increased. When the concentration of Cd²⁺ was higher than 40 μmol/L, the cell membrane of HMC3 was significantly damaged, and the damage was dose-dependent. Upon increasing the Cd²⁺ concentration, the cell morphology began to change and the adhesion also became worse. Cd²⁺ significantly increased the amount of NO released by HMC3 cells, while celastrol effectively inhibited the NO release of HMC3 cells induced by Cd²⁺. Cd²⁺ greatly increased the release of MDA in HMC3 cells, and the level of MDA decreased rapidly upon the addition of 10^-7 mol/L celastrol. Cd²⁺ increased the expression of p-PI3K protein, and the levels of p-PI3K protein and p-AKT protein were inhibited by the addition of celastrol (10^‒7 mol/L, 10^‒6 mol/L), thus preventing cell apoptosis. In conclusion, celastrol inhibits Cd²⁺ induced microglial cytotoxicity and plays a neuroprotective role.
Anti-Inflammatory Agents/pharmacology* ; Apoptosis ; Cadmium/toxicity* ; Nitric Oxide/pharmacology* ; Pentacyclic Triterpenes ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt/metabolism* ; Triterpenes/pharmacology*

OBJECTIVE: To investigate the feasibility of establishing an anterior cruciate ligament (ACL) reconstruction model using hamstring tendon autograft in cynomolgus monkeys. METHODS: Twelve healthy adult male cynomolgus monkeys, weighing 8-13 kg, were randomly divided into two groups ( n=6). In the experimental group, the ACL reconstruction model of the right lower limb was prepared by using a single bundle of hamstring tendon, and the ACL of the right lower limb was only cut off in the control group. The survival of animals in the two groups was observed after operation. Before operation and at 3, 6, and 12 months after operation, the knee range of motion, thigh circumference, and calf circumference of the two groups were measured; the anterior tibial translation D-value (ATTD) was measured by Ligs joint ligament digital body examination instrument under the loads of 13-20 N, respectively. At the same time, the experimental group underwent MRI examination to observe the graft morphology and the signal/ noise quotient (SNQ) was caculated. RESULTS: All animals survived to the end of the experiment. In the experimental group, the knee range of motion, thigh circumference, and calf circumference decreased first and then gradually increased after operation; the above indexes were significantly lower at 3 and 6 months after operation than before operation ( P<0.05), and no significant difference was found between pre-operation and 12 months after operation ( P>0.05). In the control group, there was no significant change in knee range of motion after operation, showing no significant difference between pre- and post-operation ( P>0.05), but the thigh circumference and calf circumference gradually significantly decreased with time ( P<0.05), and the difference was significant when compared with those before operation ( P<0.05). At 6 and 12 months after operation, the thigh circumference and calf circumference were significantly larger in the experimental group than in the control group ( P<0.05). At 3 and 6 months after operation, the knee range of motion was significantly smaller in the experimental group than in the control group ( P<0.05). Under the loading condition of 13-20 N, the ATTD in the experimental group increased first and then decreased after operation; and the ATTD significantly increased at 3, 6 months after operation when compared with the value before operation ( P<0.05). But there was no significant difference between the pre-operation and 12 months after operation ( P>0.05). There was no significant change in ATTD in the control group at 3, 6, and 12 months after operation ( P>0.05), and which were significantly higher than those before operation ( P<0.05). At each time point after operation, the ATTD was significantly smaller in the experimental group than in the control group under the same load ( P<0.05). The MRI examination of the experimental group showed that the ACL boundary gradually became clear after reconstruction and was covered by the synovial membrane. The SNQ at each time point after operation was significantly higher than that before operation, but gradually decreased with time, and the differences between time points were significant ( P<0.05). CONCLUSION The ACL reconstruction model in cynomolgus monkey with autogenous hamstring tendon transplantation was successfully established.
Animals ; Male ; Anterior Cruciate Ligament/surgery* ; Anterior Cruciate Ligament Injuries/surgery* ; Anterior Cruciate Ligament Reconstruction ; Hamstring Tendons/surgery* ; Knee Joint/surgery* ; Macaca fascicularis ; Transplantation, Autologous